TW201207108A - Vectors conditionally expressing therapeutic proteins, host cells comprising the vectors, and uses thereof - Google Patents

Abstract

This invention relates to the field of therapeutics. Most specifically, the invention provides methods of generating conditionally expressing vectors for one or more immunomodulators or therapeutic proteins under the control of a gene expression modulation system in the presence of activating ligand and uses for therapeutic purposes in animals. These vector may be provided to treat a variety of disorders, e.g., neoplastic disorders, infectious diseases, ocular diseases, lysosomal storage diseases, or liver diseases, through direct injection or through in vitro engineered cells, such as dendritic cells.

Description

201207108 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to the field of gene therapy for treating diseases and conditions such as cancer, lysosomal storage diseases, eye diseases, liver diseases or infectious diseases. In one embodiment, the invention provides engineered immune cells or therapeutically-supported cells (TSC) for the expression of one or more immunomodulators and the use of such cells as therapeutic agents. In another embodiment, the invention encompasses vectors which conditionally exhibit immunomodulatory agents such as IL-1 2, TNF-α disclosed herein, such as adenovirus, and methods of using such vectors. [Prior Art]

Interleukin-12 (IL-12) is a member of the first-type interleukin family involved in a variety of biological processes including, but not limited to, protective immune responses and inhibition of tumorigenesis (Abdi et al., 2006; Adorini, 1 999; Adorini, 2 0 01; Adorini et al., 2 0 0 2; Adorini et al., 1 996; Akhtar et al., 2004; Akiyama et al., 2000; Al-Mohanna et al_, 2002; Aliberti Et al., 1 9 9 6; Allavena et al., 1 994; Alii and

Khar, 2 0 0 4; Al zona et al., 1 996; Amemiya et al., 2006; Araujo et al., 2000; Arulanandam et al., 19 9 9; Athie et al., 2000; Athie -Morales et al., 2004; Bertagnolli et al., 1 992; Bhardwaj et al., 1 9 9 6; B ie derm ann et al., 2006;

Brunda and Gately, 1 994; Buchanan et al., 1 9 9 5 ; Romani et al., 1 9 9 7; Rothe et al., 1 996; Satoskar et al., 2000;

Schopf et al., 19 9 9; Thomas et al., 2000; Tsung et al., 201207108 1 997; Wolf et al., 1 9 9 4 ; Yu mi nam ochieta 1 ·, 2 0 0 7 ) The more evidence that IL-12 may be an effective target for controlling human diseases (eg, cancer), although IL-12 may be an effective cancer therapeutic because it is strong against type 1 anti-tumor NK cells, CD4+ T cells, and CD8+ T cells. Effective support activity (Trinchieri, 2003)' but recombinant human IL-12 (rhIL-12) reported toxicity in patients (Atkins et al., 1 9 9 7), together with G Μ P-class rhIL- for clinical applications The limited source of 12 makes IL-12-based treatments unsuccessful. It is therefore considered reasonable that gene therapy represents a safer and more viable treatment option. In fact, recombinant IL-12 cDNA (Heinzerling et al., 2005) or IL-based is delivered in or around the tumor with a virus (Sangro et al., 2004; Triozzi et al., 2005) or plastid-based. The first phase of clinical trials of 12 gene-modified autologous fibroblasts (Kang et al., 2001) has been considered safe and well tolerated. However, in patients with melanoma who receive these gene therapies or in patients with different cancers, there is little objective clinical response, or the clinical response is inconsistent or transient, and most of the clinical response is limited to the treatment site (Heinzerling et Al., 2005; Kang et al., 2001; Sangro et al., 2004; Triozzi et al., 2005). In cases of partial or complete remission, an increase in the number of tumor infiltrating lymphocytes was noted (Heinzerling et al., 2000; Sangro et al., 2004) and the amount of tumor-specific CD 8+ T cells in the circulation. Increased (Heinzerling et al., 2005), these findings are consistent with increased cross-sensitization of antigen-specific T cells in patients. -6- 201207108

Since DCs, which are natural but regulated sources of IL-1 2, are the most cross-sensitizing specific T cells (Berard et al., 2000), recent DC-based IL-12 gene therapy Reports of superior preclinical efficacy have received high attention (Satoh. et al., 2002; Tatsumi et al., 2003; Yamanaka et al., 2002). For example, studies have shown that intra-tumor (it) injection of intracellular (it) injections of IL-12p70-producing DCs (infected by recombinant adenovirus) results in a significant increase in broad reactivity, tumor-specific CD8+ T cell storage Cross-sensitization and tumor rejection in the library (Tatsumi et al., 2003). Considering the use of a recombinant adenovirus encoding mIL-12 under the control of the CMV promoter (rAd.cIL12, (Tatsumi et al., 2003)), the IL-12 line produced by the engineered DC is constitutive, Therefore, in terms of treatment results, the early immune effects of this interleukin in tumor lesions and the late immune effects in tumor-draining lymph nodes cannot be solved. Therefore, we need to engineer the DCs that conditionally express IL-12 in order to regulate the amount of transgene expression and activate the transgene at the appropriate time. The present invention provides promising therapeutic results using such cells. Many therapeutic proteins currently in preclinical or clinical trials are present in the patient without exhibiting deleterious adverse effects before the nucleic acid sequence is expressed by the host cell of the patient or in a suitable physiological setting. However, some proteins cause adverse effects when expressed outside of normal physiological tissues or background (e. g., exposure to non-target tissues), such as tumor necrosis factor (TNF). Systemic or even topical administration of this protein is highly toxic to many non-tumor cell types and may cause allergic reactions and cachexia. In addition, long-term exposure to TNF-α produced significantly different cellular responses than acute stimulation. For these reasons, 201207108, safe and effective TNF-α anticancer therapy is still difficult to achieve. In view of the problems associated with gene expression of genes by carrier compositions comprising proteins encoded by the nucleic acid sequences of interest, improved transport vector compositions are still needed for direct injection or for cell-based therapies. Lysosomal storage disease (LSD) represents a class of hereditary genetic diseases that are currently only treatable with protein therapeutics in the form of enzyme replacement therapy. LSD contains 49 genetic hereditary diseases characterized by defects in one or more lysosomal enzymes that cause undigested macromolecules to accumulate in lysosomes. The accumulation of these waste products causes the lysosomes in the cells to expand, resulting in cell damage and degradation. Damage that accumulates in organs and tissues causes a progressive deterioration of the physiological and/or mental state, which ultimately leads to death. Diagnosis is usually established during infancy. The severity of individual diseases varies, depending on the amount of residual enzyme activity produced by the defective gene. The incidence of LSD is about one in every 5,000 people (130,000 cases worldwide). Severity varies, depending on the amount of residual enzyme activity produced by the defective gene. Patients with severe illness may not survive more than adolescence, and less severe patients may survive to adulthood. Enzyme replacement therapy is the only method available for the treatment of L S D . Therapies include systemic infusions that target lysosomes and break down the active proteins that accumulate waste molecules. Examples of LSD protein therapeutics include Fabrazyme (Genzyme), which treats Fabry's disease, Elaprase (Shire), which treats MPSII, and Myozome, which treats Pompe's disease. (Genzyme) and Cerezyme (Genzyme), which treats Gaucher's disease 201207108. Enzyme replacement therapy has certain drawbacks, such as protein modifications that require post-translational translation, the half-life of the substitution enzyme in vivo is too short, and the patient develops an immune response to the replacement enzyme. Therefore, there is still a need in the art for enzyme replacement therapy and its replacement therapy for the treatment of lysosomal storage diseases. SUMMARY OF THE INVENTION The present invention provides a recombinant vector encoding a protein having the function of one or more immunomodulators under the control of one or more promoters. In one embodiment, the one or more promoters are conditional promoters. In another embodiment, the one or more promoters are constitutive promoters. In another embodiment, the vector encodes an adenoviral vector encoding a protein that shuts down the promoter, which can be provided by providing a soluble small molecule ligand such as a dimercaptopurine (eg, RG-115819, RG-115830, or RG). -115932) Conditional activation. This vector allows control of the expression of proteins from immune cells, T S C and from direct injection of a vector containing an immunomodulatory agent. In one embodiment, the invention provides a vector for conditionally expressing a protein having the function of one or more immunomodulators, the vector comprising a polynucleotide encoding a gene switch, wherein the polynucleotide encoding the gene switch comprises ( 1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleoside encoding a protein having one or more functions of an immunomodulator An acid that is linked to a promoter that is activated by the ligand-dependent transcription factor. In one embodiment, the immunomodulator is selected from the group consisting of -9-201207108 IL-1, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL10R DN or subunit, IL-15' IL-18, IL-21, IL-23, IL-24, IL-27, GM-CSF, IFN-α, IFN-γ, CCL3 (MIP-la) , CCL5 (RANTES), CCL7 (MCP3), XCL1 (lymphocyte chemotactic factor), CXCL 1 (MGS Α·α), CCR7, CCL 1 9 (ΜIP - 3 b), CXCL9 (MIG), CXCLIO (IP- IO) , CXCL 1 2 (SDF -1), CCL21 (6Ckine), OX40L, 4-1BBL, CD40, CD70, GITRL, LIGHT, b-defensin, HMGB 1, Flt3L, IFN-β, TNF-α, dnFADD , TGF-a, PD-L1 RNAi, PD-L1 antisense oligonucleotide, TGFbRII DN, ICOS-L, S100, CD40L, p53, survivin, p53-surplus fusion, MAGE3, PSA or PSMA. In another embodiment, the invention provides a vector for expressing a protein having the function of one or more immunomodulators and a protein having the function of IL-12, the vector comprising a polynucleotide encoding a gene switch, wherein the multinuclear The glucoside comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) encodes the function of one or more immunomodulators. a polynucleotide of a protein, and (3) a polynucleotide encoding a protein having the function of IL-12; wherein at least one of the polynucleotides of (2) and (3) is activated by the ligand-dependent transcription factor Promoter connection. In some embodiments, the vectors of the invention conditionally express TNF-a. In certain embodiments, the vector, such as an adenoviral vector, which conditionally exhibits one or more proteins having the function of an immunomodulatory agent, such as TNF-a, further comprises a nucleic acid sequence encoding a signal peptide. This signal peptide can be codon optimized -10- 201207108 . In other embodiments, the vector further comprises a 5' non-translated region (UTR), a 3' regulatory region or a second region, and enhances protein performance and/or overall yield. The invention further provides a method of producing a cell population, such as an immune cell or a TSC population, which exhibits a protein having the function of one or more immunomodulators, the method being characterized by the use of one or more immunomodulators Recombinant vectors of functional proteins are modified (eg, transfected, electroporated, etc.) such cells, wherein the vector comprises a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least one and promoter operability a ligated transcription factor sequence, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleotide encoding one or more proteins having the function of an immunomodulator, the polynucleotide being Promoter linkage of body-dependent transcription factor activation. In another embodiment, the invention provides a method of producing a population of cells, such as an immune cell or a TSC population, which exhibits a protein having one or more immunomodulatory functions and a protein having the function of IL-12,

The method modifies the cells by using a recombinant vector comprising a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription a factor sequence encoding a ligand-dependent transcription factor, (2) a polynucleotide encoding the protein having one or more functions of an immunomodulator, and (3) a polynucleotide encoding a protein having the function of IL-12; At least one of the polynucleotides of (2) and (3) is linked to a promoter activated by the ligand-dependent transcription factor. In some embodiments, the invention provides a method of increasing the performance, mRNA expression, or protein expression of an immunomodulator -11 - 201207108, such as TNF-α, the method comprising the conditional expression of one or more immunomodulatory agents A functional protein and a vector of one or more regulatory sequences, wherein the one or more regulatory sequences enhance the performance of the immunomodulatory agent, such as TNF-[alpha]. The invention further provides a cell population, such as an immune cell or a TSC population, which expresses a protein having the function of one or more immunomodulators, the cell population being recombinantly modified by a protein which conditionally exhibits the function of one or more immunomodulators (eg, transfection, electroporation, etc.), wherein the vector comprises a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one The transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleotide encoding one or more proteins having the function of an immunomodulator linked to a promoter activated by the ligand-dependent transcription factor . In another embodiment, the present invention provides a cell population, such as an immune cell or a TSC population, which exhibits a function of one or more immunomodulators and a protein having a function of 1 L. 12 A recombinant vector of a polynucleotide of a gene switch, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, (2) a polynucleotide encoding the protein having one or more functions of an immunomodulator, and (3) a polynucleotide encoding a protein having a function of 1L - 12; wherein at least (2) and (3) A polynucleotide line is linked to a promoter activated by the ligand-dependent transcription factor. In another embodiment, the invention provides a -12-12201207108 or a plurality of cells (eg, immune cells or TSCs comprising the invention) a composition of a group, wherein each cell population in the composition exhibits one or more immunomodulators not expressed by one or more of the other cell populations in the composition The immunomodulator. In one embodiment, the composition comprises two cell populations. In another embodiment, the composition comprises more than two cell populations. In another embodiment, the composition comprises three cell populations. In another embodiment, the composition comprises four cell populations. In another embodiment, the invention provides a live, in vitro engineered cell comprising a vector, such as an immune cell or TSC, the vector comprising a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) a transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleotide encoding a protein having an immunomodulatory function, the multinuclear The nucleoside is linked to a promoter that is activated by the ligand-dependent transcription factor. In another embodiment, the invention provides an in vitro engineered Q cell comprising a vector, such as an immune cell or TSC, the vector comprising a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) At least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, (2) a polynucleotide encoding a protein having an immunomodulatory function, and (3) A polynucleotide encoding a protein having the function of IL-1 2; wherein at least one of the polynucleotides of (2) and (3) is linked to a promoter activated by the ligand-dependent transcription factor. In another embodiment, the present invention provides a composition comprising a group of two or more in vitro engineered cells (eg, immune cells or TSCs) of the invention, wherein each of the compositions is The in vitro engineered cell population comprises a vector comprising a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor a sequence encoding a ligand-dependent transcription factor' and (2) a polynucleotide encoding a protein having the function of an immunomodulator, the polynucleotide being linked to a promoter activated by the ligand-dependent transcription factor, and wherein the composition The various in vitro engineered cell populations exhibit one or more immunomodulatory agents that differ from one or more of the immunomodulatory agents exhibited by other in vitro engineered cell populations in the composition. In one embodiment, the invention provides a composition comprising two or more in vitro engineered cells (eg, immune cells or TSCs), each of said populations comprising a vector comprising a multinuclear encoding a gene switch A nucleotide, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) encodes an immunomodulatory agent. a polynucleotide of a functional protein, and (3) a polynucleotide encoding a protein having the function of IL-12; wherein at least one of the polynucleotides of (2) and (3) and a ligand-dependent transcription factor thereof Activated promoter linkage. In one embodiment, the composition comprises two populations that are engineered in vitro. In another embodiment, the composition comprises more than two in vitro engineered cell populations. In another embodiment, the composition comprises three populations of cells that have been engineered in vitro. In another embodiment, the composition comprises four populations that are engineered in vitro. The invention also provides a pharmaceutical composition comprising a cell population as described herein, e.g., an immune-14-201207108 cell or a TSC population' or a composition suitable for direct injection of a cell-free population, i.e., a direct injection composition. In one embodiment, the polynucleotide encoding a protein having one or more functions of an immunomodulatory agent is under the control of a promoter of a gene switch and the polynucleotide encoding a protein having the function of IL-12 Under the control of a constitutive promoter. In another embodiment, a polynucleotide encoding a protein having an immunomodulatory function and a polynucleotide encoding a protein having a function of IL-12 are controlled at a polycistronic promoter of the gene switch under. In another embodiment, the polynucleotide encoding the protein having the function of an immunomodulator is controlled by a promoter of a gene switch and the polynucleotide encoding the protein having the function of IL-12 is linked to the gene The promoter has different conditional promoter controls. In other embodiments, the gene regulatory system of a polynucleotide encoding a protein having the function of an immunomodulator is orthogonal to a gene regulatory system of a polynucleotide having the function of IL-12. In other embodiments, the gene regulatory systems encoding individual polynucleotides of each protein are orthogonal. In one embodiment, the invention also provides treatment of cancer, such as, but not limited to, melanoma, glioma, renal cancer, prostate cancer, and the cancers listed in Table 1. IL-12 gene therapy has demonstrated anti-tumor efficacy when administered with recombinant cDNA vectors in animal model tests (Faure et al., 1 998; Sangro et al., 2005), and more particularly when genetically modified Antitumor efficacy at the time of DC administration (Satoh et al., 2002; Svane et al., 1 999; Tatsumi et al., 2003; Yamanaka et al., 2002). However, to date, the first phase of human IL-15 gene therapy using plastid or viral vectors has failed to achieve a long-lasting, objective clinical response in a cancerous environment (Heinzerling et a 1., 2 0 0 5; Kang et al., 2 00 1; S angro et al., 2004; Triozzi et al., 2005), the gene therapy described herein provides a promising therapeutic modality. In one embodiment, the invention provides a method of treating a tumor in a mammal, the method comprising the steps of: (a) intratumoral administration to a tumor microenvironment, a region surrounding the tumor, or systemic administration of the immunity of the invention a population of cells, a TSC population, or a vector (or a combination thereof) that is engineered in vitro to conditionally express one or more proteins having the function of an immunomodulatory agent: and (b) the mammal A therapeutically effective amount of an activating ligand is administered; thereby inducing a protein having the function of an immunomodulatory agent to treat the tumor. In one embodiment, the invention provides a method of treating a tumor in a mammal, the method comprising the steps of: (a) administering an immune cell or a TSC population intratumorally to a tumor microenvironment, the cell line being subjected to in vitro engineering Characterizing one or more proteins having the function of an immunomodulatory agent; and (b) administering to the mammal a therapeutically effective amount of an activating ligand; thereby inducing protein expression having the function of an immunomodulatory agent to treat the protein Tumor. In another embodiment, the invention provides a method of treating a tumor in a mammal, the method comprising the steps of: (a) administering two or more immune cells or a TSC population intratumorally to the tumor microenvironment, the cell lines In vitro engineered to conditionally express one or more of the proteins having the function of an immunomodulator, wherein the various immune cells or TSCs exhibit different one or more immunomodulators; and (b) the breastfeeding The animal is administered a therapeutically effective amount of one or more activating ligands; thereby inducing a protein having the function of an immunomodulatory agent to treat the tumor. In another embodiment, the invention provides a method of treating a tumor in a mammal, the method comprising the steps of: (a) administering an immune cell or a TSC population intratumorally to a tumor microenvironment, the cell line being ex vivo Engineering to conditionally express one or more proteins having the function of an immunomodulator and a protein having the function of IL-12, wherein at least one protein having an immunomodulatory agent or an IL-12 function is subjected to ligand-activated conditions Controlling the promoter; and (b) administering to the mammal a therapeutically effective amount of an activating ligand; thereby inducing a protein having the function of an immunomodulator and/or a protein having the function of IL-12 to treat the tumor . In another embodiment, the invention provides a method of treating a tumor in a mammal, the method comprising the steps of: (a) administering two or more immune cells or a TSC population intratumorally to the tumor microenvironment, the cell lines In vitro engineered to conditionally express one or more proteins having the function of an immunomodulator and a protein having the function of IL-12, wherein various immune cells or TSCs exhibit different functions of one or more immunomodulators a protein, wherein at least one protein having the function of an immunomodulator or IL-1 2 is under the control of a ligand-activated conditional-17-201207108 promoter; and (b) administering a therapeutically effective amount to the mammal One or more activating ligands; thereby inducing a protein having an immunomodulatory function and/or a protein having a function of IL-12 to treat the tumor. In another embodiment, the invention provides a method of treating a disease or condition in a mammal, the method comprising the steps of: U) administering to the mammal a modified population of cells, the population of cells being modified Conditionally expressing one or more proteins having the function of an immunomodulatory agent; and (b) administering to the mammal a therapeutically effective amount of an activating ligand; thereby inducing a protein having the function of an immunomodulatory agent to treat the disease or Disease. In another embodiment, the invention provides a method of treating a disease or condition in a mammal, the method comprising the steps of: (a) administering to the mammal two or more modified cell populations, the population of cells a protein that has been modified to conditionally exhibit one or more functions of an immunomodulatory agent, wherein the various modified cell populations exhibit one or more immunomodulators of different groups; and (b) the mammal is administered for treatment An effective amount of one or more activating ligands; thereby inducing a protein having the function of an immunomodulatory agent to treat the disease or condition. In another embodiment, the invention provides a method of treating a disease -18-201207108 or a condition in a mammal, the method comprising the steps of: (a) administering to the mammal a modified population of cells, the population of cells A protein that has been modified to conditionally exhibit one or more proteins having the function of an immunomodulator and a protein having the function of IL-12, wherein at least one protein having an immunomodulatory agent or an IL-1 2 function is activated by a ligand Controlling the conditional promoter; and (b) administering to the mammal a therapeutically effective amount of an activating ligand; thereby inducing a protein having the function of an immunomodulator and/or a protein having the function of IL-12 to Treat the disease or condition. In another embodiment, the invention provides a method of treating a disease or condition in a mammal, the method comprising the steps of: (a) administering to the mammal two or more modified cell populations, the population of cells A protein that has been modified to conditionally exhibit one or more proteins having the function of an immunomodulator and a protein having the function of IL-12, wherein the various modified cell populations exhibit different functions of one or more immunomodulators a protein, wherein at least one protein having the function of an immunomodulator or IL-12 is under the control of a ligand-activated conditional promoter; and (b) administering to the mammal a therapeutically effective amount of one or more activation partners The protein which induces the function of the immunomodulator and/or the protein having the function of IL-12 is thereby expressed to treat the disease or condition. The present invention also provides a method for determining the therapeutic effect of a therapy based on engineered cells such as immune cells or TSC, by measuring the amount or amount of IFN-γ expression in a subject patient at the start of treatment, thereby Producing a dose of -19-201207108, followed by administration of an effective amount of activation of a cell engineered to express one or more proteins having the function of an immunomodulator and a protein optionally having the function of IL-12 The ligand is then measured for the amount of IFN-[gamma] to produce a test amount' and the control and test amounts are compared to determine if the therapeutic formulation is effective. Also included is a method of treating a tumor, reducing tumor size, or preventing tumorigenesis in a mammal in need thereof, the method comprising (a) administering a therapeutically effective amount to the mammal to conditionally exhibit at least one immunomodulatory agent, such as IL- 12. A vector for TNF-[alpha], (b) administering to the mammal a therapeutically effective amount of one or more activating ligands, wherein the activating ligand activates the expression of a protein having the function of an immunomodulatory agent, thereby inducing immunity The protein functioning as a modulator acts to treat the tumor. In one embodiment, the invention provides a method of determining the efficacy of a therapeutic formulation based on an in vitro engineered cell, such as an immune cell or TSC, comprising: (a) measuring a first biological sample The amount or amount of interferon gamma (IFN-γ) or the amount of both, the first biological sample is obtained from the patient to be measured before administering the in vitro engineered cell, This produces a control amount; (b) administering the in vitro engineered cells to the patient to be measured 'the cells are engineered to conditionally express one or more proteins having the function of an immunomodulator and a protein optionally having the function of IL_丨2; (c) administering an effective amount of an activating ligand to the patient to be measured; -20- 201207108 (d) measuring lFN-γ in the second biological sample The amount of the expression or the amount of the activity or the amount of both, the second biological sample is obtained from the patient to be measured after administration of the in vitro engineered immune cell and the activating ligand, thereby generating a detection amount; And (e) comparing IFN-γ And detecting the amount, wherein the detection of the amount of lFN-γ expression levels, amount or activity of both increased compared to the control formulation means that the need of such treatment of effective treatment of the patient line. In one embodiment, the invention provides a method of treating a tumor, reducing tumor size, or preventing tumorigenesis in a mammal in need thereof, the method comprising: (a) administering one or more conditional expressions via intratumoral administration a vector of a protein that functions as an immunomodulator to a tumor microenvironment, the vector comprising a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein At least one transcription factor sequence encoding a ligand-dependent transcription factor, and (2) a polynucleotide encoding one or more proteins having the function of an immunomodulator activated by a ligand-dependent transcription factor Promoter operably linked, wherein the one or more immunomodulatory agents are selected from the group consisting of 11-1, 11^2, 11-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL10R DN or its subunit, IL-15, IL-18, IL-21, IL-23, IL-24, IL-27, GM·CSF, IFN-α, IFN-γ, IFN-α 1, IFN α 2, IL-15-Ra, CCL3 (MIP-la), CCL5 (RANTES), CCL7 (MCP3) ' XCL1 (lymphocyte chemotaxis Sub), CXCL1 (MGSA-a), CCR7, CCL19 (MIP-3b), CXCL9 (MIG), CXCL10 (IP-10), CXCL12 (SDF-1), CCL21 (6Ckine), OX40L, 4-1BBL, - 21 - 201207108 CD40, CD70, GITRL, LIGHT ' b-defensin, HMGB1 Flt3L, IFN-β, TNF-α, dnFADD, BCG, TGF-α, PD-RNAi, PD-L1 antisense oligonucleotide, TGFbRn DN, ICOS-L S100, CD40L, OX4 0L, p53, survivin, p53-survivin melt, MAGE3, PSA or PSMA, wherein the vector is not included in the fine; and (b) administered to the mammal A therapeutically effective amount of one or more living ligands; thereby inducing the one or more egg proteins having the function of an immunomodulatory agent to treat the tumor. The invention also provides a method of treating a disease in a non-human animal in need thereof, the method comprising: (a) administering to the non-human animal a vector for conditionally expressing white matter, the vector comprising a polynucleotide encoding a gene switch, wherein the polyglycoside The acid comprises (1) at least one transcript sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transduction factor and (2) a polynucleotide encoding one or more proteins, The nucleic acid is operably linked to a promoter activated by the ligand-dependent transcription factor, wherein the vector is not contained in the cell; and (b) the therapeutically effective amount of the one or more activating ligands is administered to the non-human; Thereby the protein is expressed to treat the disease. The invention also provides a method for treating lysosomal storage of a non-human animal in need thereof. The method comprises: (a) administering to the non-human animal a vector to express one or more proteins in a strip, the vector comprising a gene-switching switch a polyglycoside, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) encodes one or more Protein L1, cytosine, nucleoside, nucleoside, nucleoside, multi-drug, nucleus, multi--22-201207108 nuclear acid' operably linked to a promoter activated by the ligand-dependent transcription factor, wherein The vector is not included in the cell; and (b) administering to the mammal a therapeutically effective amount of - or a plurality of activating ligands; thereby inducing the one or more proteins to treat the lysosomal storage disease. The invention also provides a method of treating a liver disease in a non-human animal in need thereof, the method comprising: (a) administering to the non-human animal a vector for conditionally expressing a protein comprising: a polynucleotide encoding a gene switch, wherein the vector comprises The polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor' and (2) a polynucleotide encoding one or more proteins, The polynucleotide is operably linked to a promoter activated by the ligand-dependent transcription factor, wherein the vector is not contained in the cell; and (b) administering to the non-human animal a therapeutically effective amount of one or more activation partners The body is thereby induced to manifest the one or more proteins to treat the liver disease. Detailed description of the sequence The immunomodulator interleukin-1 (IL-1) is an important interleukin in the anti-infective inflammatory response. The polynucleotide sequences can be obtained from public databases, such as the accession number. M2 898 3 (human IL-Ια), M 1 5 3 3 0 (human IL-Ιβ), AF201830 (human IL-Ιδ), AF201831 (human IL-Ιε), AF201832 (human IL-Ιζ), AF201833 (human) IL-Ιη), NM_010554 (mouse IL-la), NM_0 08361 (mouse IL-Ιβ), NM_0 1 945 1 (mouse IL-Ιδ), NM —0 1 9450 (mouse IL-lf6), - 23- 201207108 NM_02 7 1 63 (mouse IL-lf8), N M_ 1 5 3 5 1 1 (mouse IL -1 f 9), NM_204524 (chicken IL-Ιβ), NM_017019 (rat IL-Ια) and NM_03 15 12 (rat IL-Ιβ), the sequences of which are incorporated herein by reference. Amino acid sequences of IL-1 (IL-1) can be obtained from public databases such as accession number AAA59 1 34 ( Human IL-la), AAA59 1 35 (human IL-Ιβ), AAF2 5 2 1 0 (human IL-Ιδ), AAF2 5 2 1 1 (human IL-Ιε), AAF252 1 2 (human IL-Ιζ), AAF25 2 1 3 (human IL-1 η), NP_0346 84 (mouse IL-1 a), NP_03 23 87 (mouse IL-1β), NP_062324 (mouse IL- 1 δ), NP —0623 23 (mouse IL-1 f 6), N P_ 0 8 1 4 3 9 (mouse IL-1 f 8 ), NP-705731 (mouse IL-lf9), NP_989855 (chicken IL-Ιβ), NP_058715 (rat IL-la) and NP-113700 (rat IL-Ιβ), the sequences of which are incorporated herein by reference. Laurent et al., Psychiatr Genet. 7: 103 (1997) identified polymorphic mutations in the human interleukin-1β gene. Interleukin 2 (IL-2) belongs to the interleukin family including IL-4, IL-7, IL-9, IL-15 and IL-21. The polynucleotide sequence of IL-2 is available from the public database. For example, accession numbers U25676 (human), NM_008366 (mouse), NM-204153 (chicken), and NM_053836 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of interleukin 2 (IL-2) can be obtained from public databases such as accession number AAA7 0092 (human) 'NP - 0 3 23 92 (mouse), NP - 989484 (chicken) and NP_44 62 8 8 (rat), the sequences of which are incorporated herein by reference.

Liu et al 1., Appl. Biochem. Biotechnol. 1 3 3: 77(2006) -24- 201207108 Production of mutant human IL-2, and Lorberboum et al., J. Biol. Chem. 265: 1 63 1 1 (1 990) describes the production of chimeric IL-2. Interleukin 4 (IL-4) induces the differentiation of naive T helper cells into Th2 cells, and the polynucleotide sequence of IL-4 can be obtained from a public database, such as accession number M23442 (human), NM_02 1 283 (mouse), NM_00 1 007 079 (chicken) and NM_20 1 270 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of interleukin 4 (IL-4) can be obtained from public databases such as accession number AAA5 9 1 5 0 (human), NP _0 6 7 2 5 8 (mouse), ΝΡ_0 0 1 0 0 7 0 8 0 (chicken) and NP_95 8427 (rat), their sequences are incorporated by reference

Kawashima et al., J. Med. Genet. 3 5: 502 (1 998) describes the polymorphism associated with atopic dermatitis in the IL-4 gene. Interleukin 7 (IL-7) is an important interleukin for the development of B and T cells. The polynucleotide sequence of IL-7 can be obtained from public databases such as accession number J041 56 (human), NM_0083 7 1 (mouse), Ν Μ_0 0 1 0 3 7 8 3 3 (chicken) and NM_01 3 110 ( Rats, their sequences are incorporated herein by reference. The amino acid sequence of interleukin 7 (IL-7) can be obtained from public databases such as accession number AAA59 1 56 (human), ΝΡ_0 3 2 3 9 7 (mouse), ΝΡ_00 1 032922 (chicken) and NP_037242 (rat), their sequences are included here by reference 〇

Feng et al., Genetics 1 75:545 (2007) have identified point mutations in IL-7 that lead to functional defects. Interleukin 9 (IL-9) is an interleukin produced by T cells, which is a regulator of hematopoietic -25-201207108 cells. The polynucleotide sequence of IL-9 can be obtained from public databases such as accession numbers NM_000590 (human), NM_0 0 83 73 (mouse), NM_001037825 (chicken), and NM_001105747 (rat), the sequences of which are referred to by reference. Included here. The amino acid sequence of interleukin 9 (IL-9) can be obtained from public databases such as the NP_ 000581 (human), N P_ 032399 (/J, mouse), NP_0 01032914 (chicken) and NP_00 1 09 92 1 7 (rat), the sequences of which are incorporated herein by reference. IL-12 is an interleukin that acts as a growth factor for activated T and NK cells, enhances the lytic activity of NK/lysin-activated killer cells and stimulates peripheral blood mononuclear cells (P BMC) production. Preparation of IFN-γ. The polynucleotide sequence of IL-1 2 can be obtained from public databases such as accession number NM_000882 (human IL12A), NM_002 1 87 (human IL12B), NM-0083 5 1 (mouse IL12a), NM_0083 52 (mouse IL12b) ), NM_2 1 3 5 88 (chicken IL12A), NM_2 1 3 5 7 1 (chicken IL 1 2B), NM_05 3 3 90 (rat IL12a) and NM_0226 1 1 (rat IL12b), their sequences are referred to The way is included here. The amino acid sequence of interleukin 12 (1 L-12) can be obtained from a public database, such as accession number NP_000 873 (human IL12A) 'NP — 0 0 2 1 7 8 (human IL 1 2 B ), NP_032377 ( Mouse IL12a), NP-032378 (mouse IL12b), NP-998753 (chicken IL12A), NP-998736 (chicken IL12B), NP_445842 (rat IL12a) and NP_072133 (rat IL12b), their sequences are referred to The way is included here. Interleukin 15 (IL-15) is a mediator that regulates the activation and proliferation of T and natural killer -26-201207108 cells. The polynucleotide sequence of IL-1 5 can be obtained from public databases such as accession numbers U14407 (human), NM_008 3 5 7 (mouse), EU334509 (chicken), and AF0157 19 (rat), and their sequences are The reference method is included here. The amino acid sequence of interleukin 15 (IL-15) can be obtained from public databases such as accession number AAA21551 (human), NP_03 23 83 (mouse), ABY55 3 1 2 (chicken) and AAB94 5 3 6 ( Rats, their sequences are incorporated herein by reference. Interleukin 18 (IL-18) is an interleukin produced by macrophages that induces a cellular vector immune response following infection with microbial products. The polynucleotide sequence of IL-1 8 can be obtained from public databases such as accession numbers U90434 (human), NM_008 3 60 (mouse), EU7473 3 3 (chicken) and AY2 5 8 448 (rat), The sequence is incorporated herein by reference. The amino acid sequence of interleukin 18 (IL-18) can be obtained from public databases such as accession numbers AAB500 1 0 (human), NP -0323 86 (mouse), ACE79 1 88 (chicken) and A API 4669 (Rat), their sequences are incorporated herein by reference. Interleukin 21 (IL-21) is a cytokine that induces cell proliferation to potently regulate cells of the immune system, including natural killer cells and cytotoxic T cells. The polynucleotide sequence is available from the public. A database, such as accession numbers AF254069 (human), NM_02 1 782 (mouse), NM_0 0 1 024835 (chicken), and NM - 00 1 108943 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of interleukin 21 (IL-21) can be obtained from public databases such as accession numbers AAG29348 (human), NP_068 5 54 (mouse), NP_001020006 (chicken) and NP_001102413 (rat), The sequence is incorporated herein by reference to -27-201207108. Interleukin 27 (IL-27) is an interleukin that plays an important role in regulating the activity of B and T lymphocytes. The polynucleotide sequence of IL-27 can be obtained from public databases such as accession numbers AY099296 (human), NM_145636 (mouse), and XM_344962 (rat), and their sequences are incorporated herein by reference. The amino acid sequence of (IL-27) can be obtained from public databases such as accession numbers AAM34498 (human), NP_663611 (mouse) and XP_344963 (rat), the sequences of which are incorporated herein by reference. Interferon beta 1 (IFNB 1 ) is a family member of interferon proteins that bind to specific cell surface receptors (IFNAR), which stimulate both macrophages and natural killer cells to induce antiviral responses. The nucleotide sequence can be obtained from public databases such as accession number 002 002 1 76 (human), NM_0 10510 (mouse), NM_00 1 02483 6 (chicken) and NM - 019 127 (rat), the sequences of which are referred to The way is included here. The amino acid sequence of interferon beta Ι (ΙΡΝΒΙ) can be obtained from public databases such as accession number 0020021 167 (human), ΝΡ_034640 (mouse), ΝΡ_00 1 02007 (chicken) and ΝΡ_062000 (rat), their sequences Incorporate here by reference. Interferon gamma (IF Ν - γ) is a soluble interleukin, the only type 11 interferon with antiviral, immunomodulatory and antitumor activity. The polynucleotide sequence of ΙΡΝ_γ can be obtained from public databases such as accession numbers 0006_000619 (human), NM-008337 (mouse), and ΝΜ_138880 (rat)' their sequences are incorporated herein by reference. -28- 201207108 The amino acid sequence of interferon Y (IFN-Y) can be obtained from public databases such as accession numbers NP_000610 (human), NP_032363 (mouse) and NP_620235 (rat), and their sequences are referenced. The way is included here. Tumor necrosis factor (TNF-α) is a multifunctional inflammatory cytokine secreted mainly by mononuclear spheres/macrophages, which affects fat metabolism, coagulation, insulin resistance and endothelial cell function. The nucleotide sequence can be obtained from public databases such as accession numbers X〇2910 (human), NM_01 3693 (mouse), and BC 1 0767 1 (rat), and their sequences are incorporated herein by reference 〇〇 TNF- The amino acid sequence of alpha can be obtained from public databases such as accession numbers CAA26669 (human), NP_038721 (mouse) and AAI07672 (rat), the sequences of which are incorporated herein by reference. Human TNF-α (abbreviated herein as hTNF-tx or simply hTNF) is a human interleukin with a soluble form of 17 kilodaltons (sTNF-α) and a membrane-associated form of 26 kilodaltons ( In the presence of tmTNF-α), the biologically active form consists of a trimeric body of a non-covalently bound 17 kilodalton molecule. The structure of hTNF-a is described, for example, in Pennica, D., et al. (1984) Nature 3 1 2: 724-729, Davis, J. Μ., et al. (1 987) Biochemistry 26: 1 3 22 - 1 3 26 and Jones, EY, et al. (1 9 8 9) Nature 33 8:225-22 8 〇TNF-α may be associated with TNF receptor type 1 (TNFR-1) or TNF receptor type 2 (TNFR- 2) Binding, which involves modulating immune cells, inducing apoptosis or inflammation, or inhibiting tumorigenesis or viral replication. Cell signaling ladders produced by TNF/TNFR binding are described, for example, in Wajant, H., et al. (2003) Cell Death Differ. 1 0( 1 ): 45 -65 or Chen, G., et al. 2002) -29- 201207108

Science 296: 1634-5 ° The full length human TNF-α polypeptide consists of a cytoplasmic domain, a transmembrane domain, and an extracellular domain. The polypeptide sequence of 2 3 amino acids is reported as the human TNF-α polypeptide sequence, here designated SEQ ID NO: 37, which has the cytoplasmic domain of amino acids 1 to 35 of SEQ ID NO: 37, SEQ ID NO: the transmembrane domain of amino acids 36 to 56 of 37 and the extracellular domain of amino acids 57 to 23 of SEQ ID NO: 37. SEQ ID NO: 37 is a nucleotide sequence encoding SEQ ID NO: 35 or 36. Variants of human TNF-cx include, but are not limited to, polypeptides having one or more of the following mutations: L105S, R108W, L112F, A160V, S162F, V167A, E222K, F63S, PSD 84-86VNR or E 1 83R. Chemokine hormone chemokine (C motif) ligand 1 (XCL1', also known as lymphocyte chemotactic factor), chemokates CD4 + and CD8+ T cells but does not chemoatrate mononuclear cells and induces intracellular intracellular lymphocytes The rise of calcium. The polynucleotide sequence of XCL 1 can be obtained from public databases such as accession numbers NM_002995 (human), NM_00 8 5 1 0 (mouse), and NM_1 3 43 6 1 (rat), and their sequences are incorporated by reference. Here. The amino acid sequence of XCL 1 can be obtained from public databases such as accession numbers NP-002986 (human), NP_032536 (mouse), and NP-599188 (rat), the sequences of which are incorporated herein by reference. U.S. Patent No. 6,022,534 discloses lymphocyte chemokines and their use for attracting cytotoxic T cells and/or NK cells and/or inducing intrinsic cell proliferation. Isolation and use of anti--30- 201207108 lymphocyte chemotactic factor antibody and XCL1 fusion protein also revealed that 〇CC chemokine ligand 3 (CCL3), also known as macrophage inflammatory protein 1 (M IP-1), The so-called mononuclear factor associated with the acute inflammatory state of aggregation and activation of polymorphonuclear leukocytes (a type of interleukin mainly produced by mononuclear cells and macrophages), the polynucleotide sequence of CCL3 can be obtained from public databases. , such as accession numbers NM — 0029 83 (human), NM_0 1 1 3 3 7 (mouse), and NM_0 1 3 025 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of CCL3 can be obtained from public databases such as accession numbers NP_0 02974 (human), NP_0 3 5467 (mouse), and NP_037157 (rat), the sequences of which are incorporated herein by reference. CCL5 (R ANTES) is a pro-inflammatory cytokine associated with inflammation and asthma. The polynucleotide sequence of CCL5 can be obtained from public databases such as accession number AF043 3 4 1 (human), NM_0 1 3 65 3 (mouse). And NM_031 1 16 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of CCL5 can be obtained from public databases such as accession numbers AAC03541 (human), heart _0 3 8 68 1 (mouse) and fiscal _ 112378 (rat), and their sequences are incorporated by reference. At the office. CC chemokine ligand 7 (CCL7) is a chemokine involved in macrophage aggregation during inflammation and cancer invasion. The polynucleotide sequence of CCL7 can be obtained from public databases such as accession number NM_006273 (human), NM_01 3 65 4 (mouse) and NM_001007612 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of CCL7 can be obtained from public databases such as accession number 201207108 NP-006264 (human), NP_03 86 82 (mouse) and NP_00 1 0076 1 3 (rat), and their sequences are incorporated by reference. Here. Chemokine (CXC motif) ligand 9 (CXCL9 ', also known as MIG) is a T cell chemotactic factor that can be induced by interferon gamma. The polynucleotide sequence of CXCL9 can be obtained from public databases such as accession numbers ΝΜ_0024 16 (human), ΝΜ_0 1 08 599 (mouse), and ΝΜ_145672 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of CXCL9 can be obtained from public databases such as accession numbers 002_002407 (human), ΝΡ_032625 (mouse), and ΝΡ_663705 (rat), and their sequences are incorporated herein by reference. Chemokine (C-X-C motif) ligand 10 (CXCL10) is a small interferon that functions to chemotaxis of immune system cells, adhesion of T cells to endothelial cells, anti-tumor activity, and angiogenesis. The polynucleotide sequence of CXCL10 can be obtained from public databases such as accession numbers X02 5 3 0 (human), NM_02 1 274 (mouse) and BC058444 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of the chemotactic hormone (CXC motif) ligand 10 (CXCL10) can be obtained from public databases such as accession numbers CAA2 63 70 (human), NP_067249 (mouse) and AAH58444 (rat), The sequence is incorporated herein by reference. Chemokine (CXC motif) ligand 12 (CXCL12), also known as stromal cell-derived factor 1 (SDF-1), is a small interleukin belonging to the family of mesotonin, a member of the secretin family that activates white blood cells and usually Induced by proinflammatory stimuli such as LPS, TNF or IL1. The polynucleotide sequence of CXCL12 is available from -32 to 201207108 from public databases such as accession number NM_000609 (human), ΝΜ_00 1 0 1247 7 (mouse), NM_2 04 5 10 (chicken) and NM_001033883 (rat), The sequence of such is incorporated herein by reference. The amino acid sequence of CXCL12 can be obtained from public databases such as accession number NP_000600 (human), NP-00 1 0 1 2495 (mouse), NP_98984 1 (chicken), and Ρ Ρ_0 0 1 0 2 9 0 5 5 ( Rats, their sequences are incorporated herein by reference.

Hansson et al., Microbes and Infection 8:84 1 (2006) Discuss chemokine (CC phantom) receptor 7 (CCR7) and chemokine (CC phantom) ligand 19 (CCL19, also known as ΜΙΡ-3β The interaction between them is critical to the generation of a primary immune response. The polynucleotide sequence of CCR7 can be obtained from public databases such as accession numbers 00_00 1 83 8 (human) and ΝΜ_00771 9 (mouse), and their sequences are incorporated herein by reference. The amino acid sequence of CCR7 can be obtained from public databases such as accession numbers ΝΡ_00 1 829 (human) and ΝΡ_03 1 745 (mouse), and their sequences are incorporated herein by reference. The polynucleotide sequence of CCL 19 can be obtained from public databases such as accession numbers ΝΜ_006 274 (human) and NM_0 1 1 88 8 (mouse), the sequences of which are incorporated herein by reference. The amino acid sequence of CCL19 can be obtained from public databases such as accession numbers NP_00 6265 (human) and NP_03 601 8 (mouse), and their sequences are incorporated herein by reference. CC chemokine ligand 21 (CCL21) is a clearly established ligand for CCR7, which is required for CD4+ but non-CD8+ T cells to achieve their stable state "set point", disturbance in the performance of CCL21. It is possible to change the susceptibility of autologous-33-201207108 immunity. The polynucleotide sequence of CCL2 1 can be obtained from public databases such as accession number AB002409 (human), NM_0 1 1 3 3 5 (mouse CCL21a), NM_01 1 124 (Mouse CCL21b) and NM_023 052 (mouse CCL21c); their sequences are incorporated herein by reference. The amino acid sequence of CCL2 1 can be obtained from public databases such as accession numbers BAA21817 (human), NP_03 5456 (mouse CCL21a), NP_3 5 25 4 (mouse CCL21b), and NP_075539 (mouse CCL21c), The sequences are incorporated herein by reference. Interleukin 8 (IL-8) is a chemokine, also known as neutrophil activating peptide 1 or SCYB8, which is a tissue-derived peptide secreted by several cell types in response to inflammatory stimuli. The amino acid and polynucleotide sequences of humanized anti-IL-8 antibodies are disclosed in U.S. Patent Nos. 6,13,426 and 6,1,77,98. The polynucleotide sequence of human IL-8 can be obtained from a public database, such as accession number ΝΜ_0 005 84, the sequence of which is incorporated herein by reference. The amino acid sequence of human IL-8 can be obtained from a public database, such as accession number NP-0005 75, which is incorporated herein by reference. Growth factor granules/macrophage colony-stimulating factor (GM-CSF) is an interleukin that acts as a white blood cell growth factor to stimulate stem cells to produce granules (neutrophils, eosinophils, and basophils) and Nuclear ball. The polynucleotide sequence of GM-CSF can be obtained from public databases such as accession numbers Ml 1 734 (human), NM_009969 (mouse), EU5203 03 (chicken), NM_00103 7660 (rat Csf2ra), and NM_ 1 3 3 5 5 5 (rat Csf2rb), the sequences of which are incorporated herein by reference to -34-201207108. The amino acid sequence of granule/macrophage colony-stimulating factor (GM-CSF) can be obtained from public databases such as accession numbers AAA52 1 22 (human), NP_034099 (mouse), ACB 1 1 534 (chicken), ΝΡ_00 1 032749 (rat Csf2ra) and NP_5 9 823 9 (Csf2rb), the sequences of which are incorporated herein by reference. FMS-associated tyrosine kinase ligand (FLT3/FLK2 ligand, Flt3L) can be used as a growth factor receptor on hematopoietic stem or progenitor cells or both, and the polynucleotide sequences of the other can be obtained from public databases. , such as accession numbers U04 8 06 (human) and NM_0 1 3 52 0 (mouse), their sequences are incorporated herein by reference. The amino acid sequence of FLT3/FLK2 ligand (Flt3L) is available from public data. Libraries, such as accession numbers AAA17999 (human) and NP_038548 (mouse), are included herein by reference. Transformed growth factor a (TGF-a), which is upregulated in some human cancers, can be reversely granted to the transformed phenotype to cultured cells, and the polynucleotide sequence can be obtained from a public database, such as accession number NM_00 1 09969 1 (human), NM_03 1 1 99 (mouse), NM_001001614 (chicken) and NM_012671 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of TGF-a can be obtained from public databases such as accession numbers NP_00 1 093 1 6 1 (human), NP_1 12476 (mouse), NP_001001614 (chicken) and NP_03 6 8 03 (rat), The sequence of such is incorporated herein by reference. Adjuvants -35- 201207108 Beta-defensins are antimicrobial peptides that are involved in the innate immune response against many Gram-negative and Gram-positive bacteria, fungi and viruses. The polynucleotide sequence of β-defensin can be obtained from public databases such as accession number Χ92744 (human hBD-1), AJ0001 52 (human hBD-2), AF2 1 7245 (human beta defensin 3), AJ314835 (human) β defensin 4), ΑΒ 089180 (human hBD-5), AY122466 (human defensin β 106, DEFB106), AF540979 (human beta defensin 107, DEFB107), AF529416 (human beta defensin, DEFB108), DQ012014 (human βΡ Fang Yusu 11〇, DEFB110), DQ012015 (human beta defensin 111, DEFB111), DQ012016 (human beta defensin 112, DEFB112), DQ012017 (human beta defensin 113, DEFB113), DQ012018 (human beta defensin 114, DEFB114) , DQ012019 (human beta defensin 115, DEFB115), DQ0 1 2020 (human beta defensin 116, DEFB116), DQ012021 (human beta defensin 117, DEFB117), ΝΜ_007843 (mouse defensin β 1), ΝΜ_010030 (mouse) Defensin β 2 , Defb 2 ), ΝΜ _0 1 3 75 6 (mouse defensin β 3 , Defb 3 ), NM_0 1 9728 (mouse defensin β 4 , Defb 4 ), NM_03 0734 (mouse defensin β 5 , Defb 5 ), NM_05 4074 (mouse defensin β 6, Defb6), ΝΜ_1 3 9220 (mouse defensin β 7), ΝΜ_1 5 3 1 08 ( Mouse defensin β 8 , Defb 8 ), NM_139219 (mouse defensin β 9, Defb9) and N M_ 1 3 9 2 2 5 (mouse defensin β 1 0, D e fb 1 0); The reference method is included here. The amino acid sequence of β-defensin can be obtained from public databases such as accession numbers CAA63 405 (human hBD-1), CAB65 1 26 (human hBD-2), AAF73 8 5 3 (human beta defensin 3), CAC85520 (human beta defensin 4), BAC10630 (human-36-201207108 hBD-5), AAM93908 (human defensin β 106, DEFB106), ΑΑΝ33115 (human beta defensin 107, DEFB107), AAQ0952 5 (human beta defensin , DEFB108), ΑΑΥ5 975 0 (human beta defensin 1 1 0, DEFB110), A AY 59751 (human beta defensin 111, DEFB111), AAY59752 (human beta defensin 112, DEFB112), AAY5 9753 (human beta defensin 113, DEFB113), ΑΑΥ5 9754 (human beta defensin 1 14, DEFB1 14), ΑΑΥ59755 (human beta Ρ 御 115 115, DE FB115), ΑΑΥ 59756 (human beta Ρ

素素116, DEFB116), ΑΑΥ5975 7 (human beta defensin 117, DEFB117), ΝΡ_031869 (mouse defensin β 1), ΝΡ_034160 (mouse vasopressin β 2, Defb2), ΝΡ_03 87 84 (mouse defensin β 3 ' Defb3), NP_062702 (mouse defensin β 4 , Defb4), NP_1 0965 9 (mouse defensin β 5, Defb5), NP_4734 15 (mouse defensin β 6, Defb6), NP_63 1 966 ( Mouse defensin β 7), ΝΡ_694748 (mouse defensin β 8, Defb8), ΝΡ_63 1 965 (mouse defensin β 9, Defb9) and ΝΡ_63 1 971 (mouse defensin β 10, DefblO): these The sequence is incorporated herein by reference. See also U.S. Patent No. 5,242,902 for additional human and rat defensin peptide sequences. The high-mobility group - l (HMGB1) protein acts as a non-histone chromosomal protein of interleukins, a local and systemic response of these vectors to necrotic cell death and cancer, pathogen invasion, trauma and sepsis. The polynucleotide sequence of the HMGB1 protein can be obtained from public databases such as accession numbers NM_002128 (human), NM_01 0439 (mouse), NM_204902 (chicken), and NM_0 1 2963 (rat), and their sequences are incorporated by reference. Here. The amino acid sequence of High Mobile Group-1#1^1061) can be obtained from the public database -37-201207108, such as accession number NP_002 1 1 9 (human), NP_034 5 69 (mouse), NP_990233 (chicken) And NP_037095 (rat), the sequences of which are incorporated herein by reference. The phagocytic S100 protein mediator inflammatory response and attraction of inflammatory cells to the site of tissue damage are important molecular patterns of damage-related molecular (D A Μ P) in innate immunity. See F o e 11 et al., J. Leukocyte Biol. 81:1 (2 00 6). The polynucleotide sequence of the S100 protein can be obtained from a public database, such as accession number BC014392 (human S100 Al), BC002829 (human S100 A2), BC0 1 28 93 (human S 1 00 A3), BC 0 1 6 3 0 0 (person S 1 0 0 A4), Z 1 8 95 4 (person S100D), BC00 1 43 1 (person S100 A6), BC 03 4687 (person S 100 A7), BC 00 5 92 8 (person S 1 00 A8 ), BC 0 4 7 6 8 1 (human S 1 0 0 A9), BC015973 (human S100 A10), D38583 (human calcium binding protein (calgizzarin)), N M_0 1 1 3 0 9 (mouse S 1 0 0 a 1 ), NM_009 1 1 5 (mouse SlOOb), ΝΜ_0 1 3650 (mouse S 1 00a8), NM -0091 14 (mouse S100a9), NM _ 0 1 1 3 1 0 ( /Jn mouse S100a3), NM-011311 (mouse S100a4) and NM_01 1 312 (mouse S100a5), the sequences of which are incorporated herein by reference. The amino acid sequence of the S100 protein can be obtained from public databases such as accession numbers AAH1 4392 (person S100 A1), AAH02829 (person S 1 00 A2), AAH 1 2893 (person S 100 A3), AAH1 6300 (person S 1 00 A4), CAA79479 (person S100D), ΑΑΗ0 1 43 1 (person S 100 A6), AAH3 46 87 (person S 100 A7), AAH05 92 8 (person S 1 00 A8), AAH4 7 68 1 (person S 1 00 A9), AAH15973 (human S 1 00 A 1 0), BAA07 5 97 (human calcium binding protein), NP_0 3 5 43 9 (mouse SlOOal), -38- 201207108 NP_033141 (mouse SlOOb), NP_038678 (small Murine S100a8), NP_033140 (mouse S100a9), NP-035440 (mouse S100a3), NP_035441 (mouse S100a4), and NP_035442 (mouse S100a5) 'These sequences are incorporated herein by reference. Mannan (a plant polysaccharide, a polymer of mannose) can be used to generate an immune response. U.S. Patent No. 5,807,559 discloses immunogenic conjugates of mannan which may be used to produce T cell immunity to antagonize tumor-associated carbohydrate structures or in infectious agents and/or in infected host cells. The carbohydrate structure expressed on the surface. U.S. Patent No. 5,773,425 discloses the use of mannan for relieving symptoms and/or treating toxic diseases and enhancing immune responses. BCG (attenuated live mycobacterial species) is used as a vaccine against serious and fatal tuberculosis. U.S. Patent No. 7,393,541 discloses the production of adjuvant vaccines for the production of T cell-mediated immune responses against mycobacteria in mammalian individuals. See also Hubbard and Collins, Infect. Immun. 59(2): 570. U.S. Patent No. 5,292,5 1 3 discloses a method for sterilizing BCG priming macrophages in heat in a patient in need of enhanced bactericidal and antiviral activity in vivo. The complete genomic sequence of BCG can be obtained from a public database, such as accession number NC_008769 (M. bovis BCG Pasteur strain 1173P2, complete genome). Bacterial lipopolysaccharide (LPS) is an endotoxin that induces a strong immune response when infected with Gram-negative bacteria. U.S. Patent No. 4,148,877 discloses LPS fractions derived from bacterial culture and uses the -39-201207108 component as a drug to induce resistance to bacterial infection. U.S. Patent No. 5,292,513, discloses the use of LPS to prime macrophages in a patient in need of enhanced bactericidal and antiviral activity in vivo. Costimulatory molecules (forward) 0X40 ligand (0X40L) belongs to the tumor necrosis factor (ligand) superfamily member 4 (Tnfsf4), which is expressed on dendritic cells to promote Th2 cell differentiation. The polynucleotide sequence of the 0X40 ligand can be obtained from public databases such as accession numbers X79929 (human), U12763 (mouse), and AF037067 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence 0U of the 0X40 ligand (OX40L) can be obtained from public databases such as accession numbers CAA5 62 84 (human), AAA 21871 (mouse) and AAC67236 (rat), and their sequences are incorporated by reference. Here. The 4-1BB ligand (4-1BBL) belongs to the tumor necrosis factor (ligand) superfamily member 9 (Tnfsf9), which is a type 2 transmembrane glycoprotein and is expressed on activated T lymphocytes. The polynucleotide sequence of 4-1BBL can be obtained from public databases such as accession numbers NM_0038 1 1 (human), NM_009404 (mouse), and AY3 32409 (rat), the sequences of which are incorporated herein by reference. The amino acid sequence of the 4-1BB ligand (4-1BBL) can be obtained from public databases such as accession number NP_0 03 8 02 (human), NP_03 3 43 0 (mouse) and A AQ0 1 22 8 (rat ), their sequences are incorporated herein by reference. The CD4 0 protein belongs to the tumor necrosis factor receptor superfamily member 5 and is critical for a wide range of mediators of immune and inflammatory responses, including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal-40- 201207108 Heart formation. The polynucleotide sequence of the CD40 protein can be obtained from public databases such as accession numbers 〇6〇5 92 (human), ΝΜ_1 7070 1 (mouse), ΝΜ_204665 (chicken), and ΝΜ_1 343 60 (rat), The sequence is incorporated herein by reference. The amino acid sequence of the CD40 protein can be obtained from a public database, such as the slogan CAA43 045 (human), N P_ 7 3 3 0 2 2 (/J, mouse), NP_9 8 9 9 9 6 (chicken) and NP_59 9 1 87 (rat), the sequences of which are incorporated herein by reference. CD40L (CD40 ligand or CD154) is predominantly expressed in activated T cells and is a member of the TNF molecule superfamily. It binds to CD40 on antigen-presenting cells. CD40L acts as a costimulatory molecule and induces activation of antigen-presenting cells and stimulation of T-cell receptors by MHC molecules on antigen-presenting cells. CD40L has three binding partners: CD40, α5β1 integrin and αΙΒβ3. The CD40L sequence is available from public databases such as accession numbers ΝΜ_000074 and ΜΡ_000065 (human) and ΝΜ_011616 and ΝΡ_0 3 5 74 6 (mouse). Glucocorticoid-induced tumor necrosis factor receptor family-associated protein (GITR) can cause potent tumor immunity via sputum cell stimulation. Administration of anti-GITR monoclonal antibodies (mAbs) elicits strong tumor-specific immunity and clears established tumors without inducing significant autoimmune diseases. See Ko et al., J. Exp. Med. 7: 885 (2005). The US Patent No. 6,503,1 84 B1 discloses anti-GITR antibodies. The polynucleotide sequence of the GITR ligand (GITRL) can be obtained from public databases such as accession numbers AY358868 (human) and AY359852 (mouse), the sequences of which are incorporated herein by reference. • 41 - 201207108 The amino acid sequence of the GITR ligand (GITRL) is available from public databases such as accession numbers AAQ89227 (human) and AAQ5 5265 (mouse), the sequences of which are incorporated herein by reference. The herpesvirus enters the vector factor (HVEM) binding ligand (HSVgD), also known as p30 or LIGHT, which is a member of the TNF family involved in co-stimulation with T cells. LIGHT has two receptors, the herpes virus enters the mediator (HVEM) and the lymphotoxin-beta receptor (LT-PR). In the case of HVEM ligands, HSVgD acts as a co-stimulatory factor for T cells to activate T cells, resulting in T cell proliferation and interleukin secretion. The polynucleotide and amino acid sequence of LIGHT can be found in U.S. Patent No. 7,1,8,742. U.S. Patent No. 5,654,174 describes the deletion of a variant gD protein of a carboxy terminal residue. CD70 is an interleukin that binds to CD27. It affects T cell activation. It induces proliferation of co-stimulated T cells and promotes the production of cytolytic T cells. The polynucleotide sequence of CD 70 can be obtained from public databases such as accession numbers NM_00 1 25 2 (human), NM_011617 (mouse), and ΝΜ_0 0 1 1 0 6 8 7 8 (rat), their sequences The amino acid sequence incorporated herein by reference is available from public databases such as accession numbers NP_001243 (human), NP_035747 (mouse), and NP_001 100348 (rat), and their sequences are incorporated by reference. At the office. ICOS-L is a ligand for the T cell-specific cell surface receptor ICOS as a costimulatory signal for T cell proliferation and intercellular secretion. ICOS-L also induces B cell proliferation and differentiation into plasma cells. ICOS-L plays an important role in the local tissue response of the media to inflammatory conditions. The secondary immune response can also be modulated by co-stimulation -42-201207108 Memory tau cells. The polynucleotide sequence of ICOS-L can be obtained from public databases such as accession numbers NM_0 1 5259 (human) and NM_〇1 5 790 (mouse), and their sequences are incorporated herein by reference. ICO SL Amino acid sequences can be obtained from public databases such as accession numbers NP_05 60 74 (human) and NP_05 6 605 (mouse), the sequences of which are incorporated herein by reference. The PD-L1 (aka CD274) protein is expressed in activated monocytes, T cells, and B cells. When treated with IFN-γ, the PD-L1 line in the mononuclear sphere was up-regulated, and PD-L1 in dendritic cells and keratinocytes was also up-regulated when treated with IFN-γ together with other activators. The polynucleotide sequence of the PD-L1 protein can be obtained from public databases such as accession numbers NM_014143 (human) and NM_021893 (mouse), the sequences of which are incorporated herein by reference. The amino acid sequence of the PD-L1 protein can be obtained from public databases such as accession numbers NP_05 4 8 62 (human) and NP_06 8 693 (mouse), and their sequences are incorporated herein by reference.共同 Co-stimulatory molecule (negative) Cytotoxic T lymphocyte-associated antigen 4 (CTLA4) is a member of the immunoglobulin superfamily and a costimulatory molecule expressed in activated T cells. U.S. Patent Nos. 7,34,121 and 6,984,720 disclose methods of making and using antibodies that antagonize CTLA4. U.S. Patent No. 6,984,720 also discloses the amino acid sequences of the heavy and light chains of anti-CTLA4 antibodies. A member of the PD-1 molecule immunoglobulin gene superfamily that binds to the PD-1 ligand (PD-L1). PD-L1 and the PD-1 receptor on the T cell -43-201207108 transmit a common stimulation signal to the cell to prevent the cell from continuing to the cell cycle and increase T cell proliferation. Inhibition of interactions between PD-L1 and receptors on T cells by anti-PD-L1 antibodies results in down-regulation of immune responses, termed immune cell failure. U.S. Patent No. 7,029,674 discloses a method of preparing and sequencing an anti-PD-L1 antibody. PD-L2 is mainly known as a ligand for PD-1 (or human homolog PDCD1). However, PD-L2 has been reported to be associated with a common signaling signal necessary for PD lymphocyte-independent T lymphocyte proliferation and IFN-γ production. Interaction with PDCD1 inhibits T cell proliferation and interleukin production by blocking the cell cycle. Yamazaki et al., J. of Immunol. 169: 5538 (2002) 3⁄4 Ansari et al., J. Exp. Med. 1 9 8: 63 (2003) describe the preparation of anti-PD-L2 monoclonal antibodies. Reverse immunosuppressant (tolerance inhibitor) Transgenic growth factor-p (TGF-p) is one of two types of transmembrane serine/threonine kinase receptors type I and II A multifunctional protein that interacts to regulate cell proliferation and differentiation. See Chen et al., Science 28: 1 3 3 5 (1 993). TGF receptor type II (TGFR2) phosphorylates and activates type I receptors to autophosphorize type I receptors, followed by binding and activation to SMAD transcriptional regulators. Lynch MA et al., Cancer Res. 5 8: 4227 (1 9 9 8) describes mutations in the transforming growth factor beta receptor type II gene (TGF BR2) associated with human ovarian cancer. B r an deta 1., J. Biol. Chem. 268:1 1500-1 1 5 0 3 (1 9 9 3) Describes deletion of the expected cytosolic/threonine kinase cytoplasmic domain (TGFPR2 cDNA H2-3FF Nucleotides 1172 to 2036, available from -44-201207108 public databases such as accession number M85079 and amino acid sequence such as accession number AAA6 1 164) impair all three TGF-p (l, 2 and 3) dependence Sexual gene expression. TGF-β is produced in most human tumors, which inhibit tumor antigen-specific cellular immunity. Foster et al., J. Immunother. 3 1:5 00 (2008) Describes that the expression of dominant negative TGFPR2 in cytotoxic T lymphocytes results in resistance to TGF-β inhibitory effects. TGFp synergizes with TGFa on induced transformation. It also acts as a negative autocrine growth factor. Abnormal regulation of TGFP activation and signaling may result in apoptosis. Ziyadeh et al., Proc. Natl. Acad. Sci. 97: 80 1 5 (20 00) Description The administration of anti-TGFp antibodies in db/db mice prevents renal insufficiency and glomerular sclerosis, which is evident in development. Type II diabetes model of kidney disease. Methods for the production and use of TGFp monoclonal antibodies are described in U.S. Patent No. 6,419,928. 3&1:. 611〇3-11〇££6{&1., eight 111].? 31; 11 〇 1. 1 47: 5 (1 995 ) also describes a method of producing a TGFP antibody. The amino acid and nucleotide sequence of the TGFP fusion protein construct are described in U.S. Patent No. 6,756,251. IL-10 is an interleukin produced by activated Th2 cells, B cells, keratinocytes, mononuclear cells, and macrophages. IL-10 inhibits the synthesis of a variety of interleukins, including IFN-γ' IL-2, IL-3' TNF and GM-CSF produced by activated macrophages and T helper cells. IL-10 can be used to promote the growth and differentiation of activated human B cells, inhibit the Th1 response to prevent transplant rejection and T cell-mediated autoimmune diseases. O'Farrell et al., EM B O J. 17:1006 (1998) , Kanbayashi et al., Cell

Immunol. 171:153 (1996), F ukushima et al., Br. J. -45- 201207108

Ophthalmol. 9 0 : 1 5 3 5 ( 2 0 0 6 ) v an Lent et al., Ann. Rheum. Dis. 66: 334 (2007) describes the preparation of anti-IL10 antibodies. U.S. Patent No. 7,3,26,5,67 discloses a polynucleotide sequence of an IL-10 antibody. U.S. Patent No. 5,837,232 discloses a method of treating a B cell vector autoimmune disease using an anti-IL-10 antibody. The protein of the Interleukin Signaling Inhibitor (SOCS) family forms part of a typical negative feedback system that regulates intercellular signaling. Alexander et al. Cell 98: 597 (1999) describe that interleukin signaling inhibitor 1 (SOCS1) is an important inhibitor of interferon gamma signaling, which prevents the neonatal effects of this interleukin. 1^11〇1161&1.,?1'〇£;· Natl. Acad. Sci. USA 95:1 1 4 (1 999) discusses that the SOCS1 line is associated with negative regulation of interleukin signaling via the JAK/STAT3 pathway. . Ohya et al. J. Biol. Chern. 272: 27178 (1997) describe that SOCS proteins appear to be important regulatory factors for signaling by interleukin 6 (IL-6) and leukemia inhibitory factor (LIF). U.S. Patent No. 6,5 3,277, the disclosure of which is incorporated herein by reference in its entirety, the entire entire entire entire entire entire entire entire entire entire entire entire entire entire all It is administered to the living body for therapeutic effects. Anti--3〇〇31 antibodies are also disclosed in U.S. Patent Nos. 6,323,317 and 7,049,418. A TGF-α-like cleavage polypeptide that binds to the EGF receptor and acts synergistically with TGF-β to promote non-adherent-dependent cell growth in softwood gum. Ellis et al., N. Engl. J. Med. 3 1 7:1 5 8 (1 9 8 7) Description TGF-α is associated with the associated characterization of certain tumors of melanoma. U.S. Patent No. 4,742,003 and 乂1&1161&1.,1'1^1.(^1^31〇.1^111.&-46-201207108

Cytochem. 47:949 (1 999) describes a method of preparing an anti-TGF-α antibody.

Both tumor necrosis factor receptor (TNFR1) and Fas contain a cytoplasmic Fas-associated death domain protein (FAD D), which is a Fas and TNF-induced planning cell death (apoptosis) and receptor oligomerization. It must be translated. FADD, which is named after a mammalian protein, has the ability to bind to the cytoplasmic domain or domain of the Fas receptor and inhibit the apoptosis of FAS-mediated cells. The polynucleotide sequence of F ADD can be obtained from a public database, such as registration number U2423 1, the amino acid sequence having the accession number AAA865 1 7, which is incorporated herein by reference. U.S. Patent No. 6,562,797 B1 describes a F ADD fragment or a nucleic acid encoding the same, which is a dominant negative inhibitor of a functional FADD that is functionally intact. P53 (also known as protein 53 or tumor protein 53) is a tumor suppressor protein encoded by the TP53 gene in humans. P53 is important for multicellular organisms, which regulates the cell cycle and therefore functions as a tumor suppressor associated with cancer prevention. Amino acid and polynucleotide sequences of P5 3 are available, such as accession numbers NM_00546 and NP_000537 (human) and NM_01 1 640 and NP_03 5770 (mouse). Survivin is a member of the family of apoptosis inhibitors. The function of the survivin protein is to inhibit the activation of caspase, thereby leading to the negative regulation of apoptosis or planned cell death. This function has been confirmed by disrupting the survivin-induced pathway leading to increased apoptosis and decreased tumor growth. Survivin protein is highly expressed in most human tumors and fetal tissues, but does not exist in terminally differentiated cells. This fact therefore makes survivin an ideal target for cancer treatment, because when cancer cells are targeted, normal cells are let go. The list of survivin -47- 201207108 is also regulated by the height of the cell cycle, which is only expressed in the G2-M phase. Survivors are known to be restricted to mitotic spindles by interacting with tubulin during mitosis and may play a contributing role in regulating mitosis. Regulation of survivin appears to be associated with P53 protein. Amino acid and polynucleotide sequences of p53 are available, such as accession numbers NM_00 1 0 1 2270 and NP_001012270 (human) and NM_001012273 and NP_001012273 (mouse) 0 melanoma-associated antigen 3 (MAGE3) amino acid sequence As shown in registration number P43357-l (UniParc). Prostate specific antigen (PSA) is a protein produced by cells of the prostate. PSA is present in small amounts in the serum of people with prostate health, but it is usually elevated in people with prostate cancer and other prostate diseases. Prostate-specific membrane antigen (PS Μ A) is a type 2 integral membrane glycoprotein found in prostate tissue and some other tissues. This is a possible therapeutic target for prostate cancer. BRIEF DESCRIPTION OF THE SEQUENCE LISTING SEQ ID NO: 1 is a polynucleotide sequence encoding a construct of mlL-12 and m-IL21. SEQ ID NO: 2 is a polynucleotide sequence encoding a construct of hIL-12 and h-IL21. SEQ ID NO: 3 is a polynucleotide sequence encoding a construct of mlL-21 and m-IL15. SEQ ID NO: 4 is a polynucleotide sequence encoding a construct of mlL-12 -48-201207108 SEQ ID NO: 5 is a polynucleotide sequence encoding a construct of hIL-21 and hIL-15. SEQ ID NO: 6 is a polynucleotide sequence encoding a construct of hIL-21. SEQ ID NO: 7 is a polynucleotide sequence encoding a construct of mIL-21. SEQ ID NO: 8 is a construct encoding hIL-21. The polynucleotide sequence of SEQ ID NO: 9 is a polynucleotide sequence encoding mIL-21. SEQ ID NO: 10 is the amino acid sequence of mIL-21. SEQ ID NO: 11 is a polynucleotide sequence encoding mIL-15. SEQ ID NO: 12 is the amino acid sequence of mIL-15. SEQ ID NO: 13 is a polynucleotide sequence encoding mp40 of mIL-12 〇 SEQ ID NO: 14 is the amino acid sequence of mp40 of mIL-12. SEQ ID NO: 15 is the polynucleotide sequence encoding mp35 of mIL-12 〇 SEQ ID NO: 16 is the amino acid sequence of m35 of mlL-12. SEQ ID NO: 17 is a polynucleotide sequence encoding hIL-21. SEQ ID NO: 18 is the amino acid sequence of hIL-21. SEQ ID NO: 19 is a polynucleotide sequence encoding hIL-15. SEQ ID NO: 20 is the amino acid sequence of hIL-15. SEQ ID NO: 21 is the polynucleotide sequence encoding p40 of hIL_i2. -49- 201207108 SEQ ID NO: 22 is the amino acid sequence of p40 of hIL-12. SEQ ID NO: 23 is the polynucleotide sequence encoding p35 of hIL-12. SEQ ID NO: 24 is the amino acid sequence of P3 5 of hIL-12. SEQ ID NO: 25 is the nucleic acid sequence of an ephedrine response element found in Drosophila. SEQ ID NO: 26 is the nucleic acid sequence of a cutaneous hormone response element found in Drosophila melanogaster. SEQ ID NO: 27 is the nucleic acid sequence of an ecdysone response element found in Drosophila melanogaster. SEQ ID NO: 28 is a restriction site for homing endonuclease (HE) enzyme (I-Scel). SEQ ID NO: 29 is the DNA sequence of an adenoviral vector comprising the human IL-12 coding sequence: Ad-RTS-hIL-12 (SP1-Rhe〇IL-12). SEQ ID NO: 30 is the nucleic acid sequence of human TNF wild type 5' UTR. SEQ ID NO: 31 is the nucleic acid sequence of 5U2 5' UTR. SEQ ID NO: 32 is a codon-optimized nucleic acid sequence encoding an IL-2 signal peptide. SEQ ID NO: 3 3 is a wild-type nucleic acid sequence encoding a human TNF-α signal peptide. SEQ ID NO: 3 4 is a codon-optimized nucleotide sequence encoding a human TNF-a signal. SEQ ID NO: 35 is a wild-type nucleic acid sequence encoding human TNF-a. SEQ ID NO: 3 6 is a codon-optimized nucleic acid sequence encoding human TNF-a. -50- 201207108 SEQ ID NO: 3 7 is the amino acid sequence of human TNF-ct. SEQ ID NO: 38 is the nucleic acid sequence of the 3' regulatory region comprising a nucleotide sequence encoding a SV40 polyadenylation signal. SEQ ID NO: 39 is a nucleic acid sequence of a regulatory region comprising a nucleotide sequence encoding a human growth hormone polyadenylation signal. SEQ ID NO: 40 is a nucleic acid sequence comprising the wild type human TNF-α 3' UTR. SEQ ID NO: 41 is the nucleic acid sequence of a human TNF-α 3' UTR AtoC mutant. SEQ ID NO: 42 is the nucleic acid sequence of human GAST 3'UTR. SEQ ID NO: 43 is the nucleic acid sequence of a synthetic V regulatory region. SEQ ID NO: 44 is the nucleic acid sequence of human GAPDH 5' UTR. SEQ ID NO: 45 is a wild-type nucleic acid sequence of insulin SP. SEQ ID NO: 46 is a wild-type nucleic acid sequence encoding a human FGF-19 signal peptide. SEQ ID NO: 47 is the nucleic acid sequence of vector 433 1 8 . SEQ ID NO: 48 is the nucleic acid sequence of vector 43319. SEQ ID NO: 49 is the nucleic acid sequence of vector 43 3 20 . SEQ ID NO: 50 is the nucleic acid sequence of vector 43321. SEQ ID NO: 5 1 is the nucleic acid sequence of vector 43322. SEQ ID NO: 5 2 Nucleotide sequence of vector 43 3 23 . SEQ ID NO: 53 is the nucleic acid sequence of vector 43324. SEQ ID NO: 54 is the nucleic acid sequence of vector 43325. SEQ ID NO: 55 is the nucleic acid sequence of vector 43326. -51 - 201207108 SEQ ID NO: 56 Nucleotide sequence of vector 43327. SEQ ID NO: 5 7 Nucleotide sequence of vector 4332. SEQ ID NO: 58 is the nucleic acid sequence of vector 43329. SEQ ID NO: 59 is the nucleic acid sequence of vector 43533. SEQ ID NO: 60 Nucleotide sequence of vector line 43 5 3 4 . SEQ ID NO: 61 is the nucleic acid sequence of the vector VVN2823 (Ad-RTS-hIL-12). SEQ ID NO: 62 is the nucleic acid sequence of the vector VVN25 3 9 (Ad-RTS-mIL-12). DETAILED DESCRIPTION OF THE INVENTION Definitions All technical terms, descriptions, and other scientific terms or phrases used herein are intended to have the meanings In many instances, terms that have a commonly understood meaning are defined herein for clarity and/or ease of reference and understanding, and the inclusion of such definitions herein should not be construed as a representative of the field. The difference in substance. A general understanding of the terms and/or methods and/or protocols of molecular biology can be found in Rieger et al., Glossary of Genetics: Classical and Molecular, 5th edition, Springer-Verl ag: New York, 1991, Le win, Genes V. , Oxford University Press: New York, 1 994 ' Sambrook et al.? Molecular Cloning, A Laboratory M anu al (3 d ed . 2001 ) and Ausubel et al., Current Protocols in Molecular 201207108

Biology (1994). Where appropriate, methods involving the use of commercially available kits and/or reagents are generally performed according to the manufacturer's instructions and/or protocols and/or parameters, unless otherwise stated. For the purposes of the present invention, the term "isolated" means a substance that has been removed from the original environment of the biological substance (cell, nucleic acid or protein) in which the substance naturally occurs. For example, a polynucleotide that is present in a plant or animal in its native state is not isolated, whereas the same polynucleotide that is separated from adjacent nucleic acids in the environment in which the polynucleotide is naturally found is considered "isolated." of. When used to refer to a biological substance, the term "purified" does not require that the substance be present in a form that exhibits absolute purity and excludes the presence of other compounds. It is instead a relative definition. "Nucleic acid", "nucleic acid molecule", "oligonucleotide", "nucleotide" and "polynucleotide" are used interchangeably. These terms refer to ribonucleosides (adenosine) in single-stranded or double-stranded helix. , guanosine, uridine or cytidine; "RN A molecule") or deoxyribonucleoside (deoxyadenosine, deoxyguanosine, deoxyuridine or deoxycytidine; "DNA molecule") phosphoric acid Ester-polymerized form, or any of these phospholipid analogs, such as phosphorothioates and thioesters. Double-stranded DNA-DNA, DNA-RNA and RNA-RNA helix are possible. Molecular nucleic acid molecules, especially DNA or RNA molecules It refers only to the primary and secondary structure of the molecule, and does not limit it to any particular tertiary form. Therefore, this term includes, inter alia, linear or circular DNA molecules (eg, restriction fragments), plastids, supercoiled DNA, and A double-stranded DNA found in a chromosome. When discussing the structure of a specific double-stranded DNA molecule, it may be based on the sequence of 5' to -53-201207108 3 ' of the non-transcribed strand of the DNA that is normally used (ie, having a sequence of mRN A homologous sequences) to describe the sequence herein. "Recombinant DNA Child "means the suffering of the DNA molecule molecular biological manipulation. DNA includes, but is not limited to, cDNA, genomic DNA, plastid DNA, synthetic DNA, and semi-synthetic DNA. The term "fragment" when used in reference to a polynucleotide sequence refers to a nucleotide sequence that is reduced relative to the length of the reference nucleic acid and which comprises, in addition to the common portion, a nucleotide sequence that is identical to the reference nucleic acid. For the purposes of the present invention, the nucleic acid fragment may be included within the larger polynucleotide when it is likely to be a component of a larger polynucleotide. The fragments comprise at least 6, 8, 9, 10, 12, 15, 18, 20, 21, 22, 23, 24, 25, 30, 39, 40, 42, 45, 48, 50 of the nucleic acids of the invention. 51, 54, 57, 60, 63, 66, 70, 75, 78, 80, 90, 100, 105, 120, 135, 150, 200, 300, 500 ' 720 ' 900 ' 1000 , 1500 , 2000 , 3000 , Oligonucleotides of 4000, 5,000 or more contiguous nucleotides, or consist of such oligonucleotides. As used herein, "isolated nucleic acid fragment" refers to a polymer of single or double stranded RNA or DNA, which may optionally comprise synthetic, non-natural or altered nucleotide bases. An isolated nucleic acid fragment in the form of a DN A polymer may comprise one or more segments of cDNA, genomic DNA or synthetic DNA. "Gene" refers to a polynucleotide comprising a nucleotide encoding a functional molecule, including a transcriptional producer only (eg, a biologically active RN A species) or a transcriptional and translational producer (eg, a polypeptide). The term "gene" encompasses cDNA and genomic DNA nucleic acids. "Gene" also refers to a nucleic acid fragment that expresses a particular RNA, -54-201207108 protein or polypeptide, including regulatory sequences preceding (5, non-coding sequences) and after (3' non-coding sequences) of the coding sequence. "Native gene" refers to a gene found in nature that has its own regulatory sequence. "Chimeric gene" refers to any gene of a non-native gene that contains regulatory and/or coding sequences that are not found together in nature. Thus, a chimeric gene may comprise regulatory sequences and coding sequences derived from different sources, or regulatory sequences and coding sequences derived from the same source but arranged in a manner different from that found in nature. A chimeric gene may comprise coding sequences derived from different sources and/or regulatory sequences derived from different sources. "Endogenous gene" means a natural gene whose natural location is within the genome of an organism. A "foreign" gene or a "heterologous" gene refers to a gene that is not normally found in the host organism but is introduced into the host organism by gene transfer. The foreign gene may comprise a native gene inserted into a non-native organism, or a chimeric gene. "GMO" is a gene that is introduced into the genome via a transformation program. For example, the interleukin-12 (IL-12) gene encodes an IL-12 protein. IL-12 is a heterodimer of 35 kilodalton sub-q (P35) and 40 kilodalton subunit (p40), which are linked via disulfide bonds to form a fully functional IL- 12p70. The IL-12 gene encodes two subunits, p35 and p40. "Heterogenous DNA" refers to DNA that is not naturally found in the cell or in the part of the cell. Heterologous DNA may contain genes that are foreign to the cell. The term "genome" includes not only chromosomal DNA or RNA, but also mitochondria, chloroplasts, and viral DNA or RNA. When a single-stranded form of a nucleic acid molecule such as cDNA, genomic DNA or -55-201207108 RN A can bind to other nucleic acid molecules at an appropriate temperature and solution ionic strength, the nucleic acid molecule can "hybridize" with another nucleic acid molecule. . Hybridization and washing conditions are well known and described in Sambrook et al. in Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harb〇r (1 989), in particular Chapter 11 and Table 11.1. The conditions of temperature and ionic strength determine the "stringency" of hybridization. Stringency conditions can be adjusted to screen for fragments of similar degree, from reluctance to similar fragments (such as homologous sequences derived from distantly related organisms) to highly similar fragments (such as from closely related organisms for replication functionality) Enzyme gene). For preliminary screening of homologous nucleic acids, low stringency hybridization conditions corresponding to Tm 5 5 ° C can be used, such as 5 times S S C, 0.1% S D S, 0.2 5 °/. Milk and no formamide; or 30% methotrexate, 5 times S S C, 0.5% SDS. Moderate stringency hybridization conditions correspond to higher Tm, such as 40% methotrexate and 5 or 6 times SSC. Highly stringent hybridization conditions correspond to the highest Tm, such as 50% methotrexate and 5 or 6 times S S C. Hybridization requires that the two nucleic acids must contain complementary sequences, but depending on the stringency of the hybridization, there may be a mismatch between the bases. The term "complementary" is used to describe the relationship between nucleotide bases that can hybridize to each other. For example, in the case of DN A, the adenine is complementary to thymine, and the cytosine is complementary to guanine. Thus, the invention also includes isolated nucleic acid fragments that are complementary to the complete sequences disclosed or used herein, as well as substantially similar nucleic acid sequences. In one embodiment of the invention, the polynucleotide is detected by the use of hetero-56 - 201207108, the hybridization conditions being included in the Tm 55 °C hybridization step and using conditions as described above. In other embodiments, the Tm is 60 ° C, 63 ° C or 65 ° C. Cleaning after hybridization also determines the conditions of stringency. A set of conditions using a series of washing steps, initially washed at room temperature with 6 times SSC, 0.5% SDS for 15 minutes (min), followed by repeated washing at 2 times SSC, 0.5% SDS for 30 minutes at 45 ° C, and then The washing was repeated twice at 50 ° C for 0.2 minutes with 0.2 times SSC and 0.5% SDS. A set of stringency conditions used higher temperatures, with each cleaning step being the same as above, except that the temperature was increased to 60 °C for the last two times with 〇·2 times SSC, 0.5% SDS cleaning for 30 minutes. Another set of highly stringent conditions for the last two washes at 65 ° C with 0.1 times SSC, 0.1% SDS cleaning 〇 nucleic acid hybridization appropriate rigor depends on the length of the nucleic acids and the degree of complementarity, these are well known in the field Variation. The higher the degree of similarity or homology between the two nucleotide sequences, the higher the hybrid of the nucleic acid having the sequences. The relative stability of nucleic acid hybridization (corresponding to a higher Tm) is decremented in the following order: RNA: RNA, DNA: RNA, DNA: DNA. For hybrids longer than 100 nucleotides, the equation for calculating Tm has been developed (see Sambrook et al., supra, 9.50-0.51). In the case of hybridization of shorter nucleic acids (i.e., oligonucleotides), the position of the mismatch is more important, and the length of the oligonucleotide determines its specificity (see Sambrook et al., supra, 11.7-11.8). In one embodiment of the invention, the 'polynucleotide is detected by hybridization conditions comprising a hybridization step at a concentration of less than 500 millimolar salt - 57 - 201207108 and at least 37 ° C, And at least 63. (The temperature is 2 times the washing step of SSPE is used. In another embodiment, the hybridization conditions comprise a hybridization step of less than 200 millimolar salt concentration and at least 37 ° C. In other embodiments The hybridization conditions comprise 2 times SSPE and a hybridization and washing step at 63 ° C. In another embodiment, the hybridizable nucleic acid is at least about 1 nucleotide in length. Preferably The shortest length of the hybridized nucleic acid is at least about 15 nucleotides 'e.e., at least about 20 nucleotides, such as at least 30 nucleotides. Further, the skilled artisan will understand the temperature and the wash solution salt. The concentration can be adjusted as needed depending on various factors such as the length of the probe. The term "probe" refers to a single-stranded nucleic acid molecule that can be base paired with a complementary single-stranded target nucleic acid to form a double-stranded molecule. The term used herein" "Oligonucleotide" refers to a short nucleic acid that can hybridize to a genomic DNA molecule, cDNA molecule, plastid DNA or mRNA molecule. Oligonucleotides can be covalently shared, for example, with 32P-nucleotides or labeled (such as biotin). The nucleotide of the yoke is labeled. Oligonucleotides can be used as probes to detect the presence of nucleic acids. Oligonucleotides (one or both of which can be labeled) can be used as PCR primers for cloning nucleic acids The full length or fragment is sequenced for DN A or used to detect the presence of a nucleic acid. Oligonucleotides can also be used to form a triple helix with a DN A molecule. Typically, oligonucleotides are prepared synthetically, preferably Synthesizing on a nucleic acid synthesizer. Thus, the prepared oligonucleotide may have a non-naturally occurring phosphoester bond analog, such as a thioester bond, etc. "Introduction" refers to hybridization with a target nucleic acid sequence to produce a double-stranded nucleic acid. Region-58-201207108 Oligonucleotide' This double-stranded nucleic acid region can be used as a starting point for DN A synthesis under appropriate conditions. These primers can be used for polymerase chain reaction or DN A sequencing. The enzyme linkage reaction is abbreviated as PCR, which refers to an in vitro method of amplifying a specific nucleic acid sequence by the action of an enzyme. PCR involves repeated cycles of temperature cycles. Each cycle contains three stages: denaturation of the template nucleic acid to separate the target molecules. Single-strand PCR oligonucleotide Binding to the template nucleic acid and extending the adhesion primer by DNA polymerase. PCR provides detection of the presence or absence of the target molecule, and determines the relative amount of the target molecule in the nucleic acid starting pool under quantitative or semi-quantitative conditions. The "reverse transcription polymerase chain reaction" is abbreviated as RT-PCR, which refers to the production of a target cDNA molecule from an RNA molecule by the action of an enzyme, and then amplified by the enzyme as described above in the target cDNA molecule. An in vitro method for a specific nucleic acid sequence. RT-PCR also provides a method for detecting the presence or absence of a target molecule and determining the relative amount of the target molecule in the nucleic acid starting pool under quantitative or semi-quantitative conditions.

V DN A "coding sequence" or "coding region" refers to a double-stranded DN A sequence encoding a polypeptide, and when the "coding sequence" is placed under the control of an appropriate regulatory sequence, it can be intracellular or ex vivo (ex vivo) ), in vitro or in vivo (in vi vo) is transcribed and translated into a polypeptide. "Appropriate regulatory sequence" refers to a nucleotide sequence located upstream of a coding sequence (5' non-coding sequence), within a coding sequence, or downstream of a coding sequence (3' non-coding sequence), which sequence affects transcription of the relevant coding sequence , RNA processing or stability or translation. Regulatory sequences can include promoters, translation leader sequences, introns, polyadenylation recognition sequences -59-201207108, RN A treatment sites, effector binding sites, and stem-loop structures. The boundaries of the coding sequence are defined as the start codon at the 5' (amino) terminus and the translation stop codon at the 3' (carboxy) terminus. A coding sequence can include, but is not limited to, a prokaryotic sequence, a cDNA derived from mRNA, a genomic DNA sequence, and even a synthetic DNA sequence. If the coding sequence is intended for use in eukaryotic expression, a polyadenylation signal and a transcription termination sequence will typically be present at the 3' end of the coding sequence. "Open reading frame" is abbreviated as ORF and refers to a nucleic acid sequence (whether DNA, cDNA or RNA) containing a translation initiation signal or a start codon (such as ATG or AUG) and a stop codon, and may be Translated into a polypeptide sequence. The term "head-to-head" as used herein describes the directionality of the relationship of two polynucleotide sequences to each other. When the 5' end of the coding strand of one polynucleotide is adjacent to the 5' end of the coding strand of another polynucleotide, the two polynucleotides are arranged head-to-head in order to make each polynucleoside The direction of acid transcription proceeds toward the 5' end away from the other polynucleotide. The term "head-to-head" can be abbreviated as (5,)_ to _(5'), and can also be represented by the symbol - 〇 or (y - 5'5' - 31). The term "tail-to-tail" as used herein describes the directionality of the relationship of two polynuclear acid sequences to each other. When a polynucleotide encodes a strand; 3, the end is adjacent to the 3' end of the encoded strand of another polynucleotide, and the two polynuclear acid acids are arranged in a tail-to-tail direction. The transcriptional direction of the polynucleotide proceeds toward the other-polynucleotide. The term "tail-to-tail" can be abbreviated as (3,)_for _(3,), or symbol () or (5'-3'3'-51). The term "head-to-tail" as used herein describes the directionality of the relationship of two polynucleotide sequences to each other. When the 5, end of the coding strand of one polynucleotide is adjacent to the 3' end of the other strand of the -60-201207108-polynucleotide, the two polynucleotides are arranged in a head-to-tail orientation. The transcription direction of each polynucleotide is advanced in the same direction as the other polynucleotide. The term "head-to-tail" can be abbreviated as (5·)-to-(3'), or symbolic) or (5·-3'5'-3'). The term "downstream" refers to the nucleotide sequence located 3' to the reference nucleotide sequence. In particular, the downstream nucleotide sequence is usually associated with a sequence after the start of transcription. For example, the translation initiation codon of a gene is located downstream of the transcription initiation site. The term "upstream" refers to a nucleotide sequence located 5' to a reference nucleotide sequence. In particular, the upstream nucleotide sequence is typically associated with a sequence located on the 5' side of the coding sequence or at the transcription initiation site. For example, most promoters are located upstream of the transcription start site. The terms "restricted endonuclease" and "restriction enzyme" are used interchangeably and refer to an enzyme that binds to a particular nucleotide sequence within a double strand of DN A and cleaves within that particular nucleotide sequence. "Homologous recombination" refers to the insertion of a foreign DNA sequence into another DNA molecule, such as insertion of a vector into a chromosome. Preferably, the vector targets a specific chromosome site for homologous recombination. In the case of specific homologous recombination, the vector will comprise a region of sufficient length to be homologous to the chromosomal sequence to allow for complementary binding and for inclusion of the vector in the chromosome. The longer the region of homology and the higher the degree of sequence similarity, the greater the efficiency of homologous recombination. Some methods known in the art can be used to propagate the polynucleotides of the present invention. Once the appropriate host system and growth conditions are established, the recombinant expression vector can be propagated and prepared in large quantities. As used herein, the expression -61 - 201207108 vectors include, but are not limited to, the following vectors or derivatives thereof: human or animal viruses such as vaccinia virus or adenovirus; insect viruses such as baculovirus; yeast vectors; phage A carrier (eg, λ) and a plastid and viscous DNA vector, etc., refer to any carrier used to clone and/or transfer a nucleic acid into a host cell. The vector may be a replicon that can be attached to another DN Α fragment to facilitate replication of the attachment segment. "Replicon" refers to any genetic element (eg, plastid, phage, plastid, chromosome, virus) that functions as an autonomous unit of DNA replication in vivo (ie, self-regulated replication). The term "vector" includes both viral and non-viral vehicles for introducing nucleic acids into cells in vitro, in vitro or in vivo. Many vectors known in the art can be used to manipulate nucleic acids, incorporate reaction elements and promoters into genes, and the like. Possible vectors include, for example, plastid or modified viruses including, for example, bacteriophages such as lambda derivatives, or plastids such as PBR3 22 or pUC plastid derivatives, or Bluescript vectors. Another example of a carrier that can be used in the present invention is 1114 & 76 (^〇): 1^Production System (Blacksburg, VA) Inchson, as described in WO 2007/038276 (Intrexon) Inc. For example, the insertion of a DNA fragment corresponding to a reaction element and a promoter into an appropriate vector can be accomplished by ligating the appropriate DN A fragment to a selection vector having a complementary viscous end. The ends of the DN A molecules may be enzymatically modified or may be joined to the ends of the DNA by ligation of a nucleotide sequence (linker) to create any site. Such vectors may be engineered to contain a selectable marker gene, This allows screening of cells that have been incorporated into the genome of the cell. These markers allow for the identification and/or screening of host cells that incorporate and express the protein encoded by the marker -62-201207108. Viral vectors (especially retroviruses) Vectors have been used in various 'intracellular gene delivery applications' and are also used in living organisms. Viral vectors that can be used include, but are not limited to, retroviruses, adeno-associated viruses, Prion, baculovirus, vaccinia virus, herpes simplex virus, EB (Epstein-Barr) virus, adenovirus, geminivirus and cauliflower mosaic virus vector. Non-viral vectors include plastids, liposomes, charged lipids (cell turnover) Ingredients, DNA-protein complexes, and biopolymers. In addition to nucleic acids, the vector may contain one or more regulatory regions, and/or may be used to select, measure, and monitor nucleic acid transfer results (to which tissue, A selectable marker for a period of expression, etc. The term "plastid" refers to an extrachromosomal element that normally carries a gene that is not part of the metabolism of the cell center, and is usually in the form of a circular double-stranded DN A molecule. Sites that replicate sequences, genomic embedded sequences, phage or nucleotide sequences, linear, circular or supercoiled single or double stranded DNA or RNA from any source, many of which have been joined or reconstituted uniquely Constructor's ability to introduce the promoter fragment of the selected gene product and the DN A sequence into the appropriate 3 · non-translated sequence 〇 "cloning (clon "inger" is a "replicon" which is a nucleic acid (preferably DNA) that is sequentially replicated and contains a unit length of the origin of replication, such as a plastid, bacteriophage or plastid 'cloning vector and another nucleic acid region Segment attachment to facilitate replication of the attachment segment. The cloning vector can be replicated in one cell type and expressed in another cell type ("shuttle vector"). The cloning vector can be -63-201207108 containing one or more A sequence for screening a cell comprising the vector and/or one or more multiplex cloning sites for insertion of a sequence of interest. The term "expression vector" refers to a vector designed to express an inserted nucleic acid sequence, Plastid or carrier. The cloned gene, i.e., the inserted nucleic acid sequence, is typically placed under the control of a control element, such as a promoter, minimal promoter, enhancer, or the like. The initial control regions or promoters that can be used to drive the expression of a nucleic acid in a desired host cell are numerous and familiar to those skilled in the art. Almost any promoter that drives the expression of these genes can be used in expression vectors including, but not limited to, viral promoters, bacterial promoters, animal promoters, mammalian promoters, synthetic promoters, constitutive promoters, tissue-specific Promoter, pathogenesis or disease-related promoter, development-specific promoter, inducible promoter, light-regulated promoter; CYC1, HIS3, GAL1, GAL4, GAL 10, ADH1, PGK, PH05, GAPDH, ADC1, TRPI, URA3 'LEU2' ENO, ΓΡ/, alkaline phosphatase promoter (can be used in the performance of beer brewing); JOZi promoter (can be used in Pichia pastoris); β-endoaminase, /ac, set, iei, irp, //hu and Γ7, iac and ircr promoters (can be used in E. coli (five co/i)); light-regulating-, seed-specific - 'pollen specific -, ovarian specific -, cauliflower mosaic virus 35S, minimal CMV 35S, cassava vein mosaic virus (CsVMV), chlorophyll a/b binding protein, ribulose 1,5 -bisphosphate carboxylase, stem Sex, root specificity, chitinase, stress-induced, rice tungro baculovirus, plant super-promoter, potato lysamine peptidase, nitrate reductase, -64- 201207108 mannopine synthesis Enzymes, nopaline synthase, ubiquitin, zein and anthocyanin promoters (used for expression in plant cells); animal and mammalian promoters known in the art include, but are not limited to, the SV40 early (SV40e) promoter Promoter of the 3' long terminal repeat (LTR) of the Rous sarcoma virus (RSV), the promoter of the adenovirus (Ad) E1A or the major late promoter (MLP) gene, cytomegalovirus (CMV) Early promoter, herpes simplex virus (HSV) thymidine kinase (TK) promoter, baculovirus IE1 promoter, elongation factor la (EF1) promoter, phosphoglycerate kinase (PGK) promoter, ubiquitin (Ubc) Promoter, albumin promoter, regulatory sequence of mouse promoter and transcriptional control region, metallothionein-L promoter and transcriptional control region, universal promoter (HPRT, vimentin, α-actin, microtubule) Protein and the analog), medium Promoter of mesenchymal (intermuscular protein, neurofilament, keratin, GFAP and the like), promoter of therapeutic gene (MDR, CFTR or factor VIII and the analog), pathogenesis or disease a related promoter, and a promoter that exhibits tissue specificity and has been used in gene transfer animals, such as an elastase I gene control region active in pancreatic vesicle cells; an active insulin gene control region in pancreatic β cells An immunoglobulin gene control region active in lymphoid cells, a mouse mammary tumor virus control region active in the sputum, breast, lymphoid and obese cells; an active albumin gene in the liver sputum, Αρο ΑΙ and Apo All control region, active α-fetal protein gene control region in liver, α 1 _ antitrypsin gene control region active in liver, β-globulin gene control active in bone marrow cells Region, the myelin basic protein gene control region active in the oligodendrocytes of the brain, the myosin light chain-2 gene control in the skeletal muscle -65-201207108 region and in the hypothalamus With activity of gonadotropin releasing hormone gene control region, pyruvate kinase promoter, villin promoter, promoter of the fatty acid binding intestinal protein promoter, smooth muscle cell α - actin promoter, the proteins and the like. Furthermore, these expression sequences may be modified by the addition of enhancers or regulatory sequences and the analogs. The vector can be introduced into a desired host cell by methods known in the art, such as transfection, electroporation, microinjection, transduction, cell fusion, DEAE dextran, calcium phosphate precipitation, liposome transfection (lysosome). Fusion), using a gene gun or DNA vector transporter (see, eg, Wu et al., J Biol. Chem. 267:963 (1 992), Wu et al., J Biol. Chem. 263:1 462 1 (1 9 8 8) and Hartmut et al., Canadian Patent Application No. 2,012,311). The polynucleotide of the present invention can also be introduced into a living body via lipofection. In the past decade, liposomes have been increasingly used to encapsulate and transfect nucleic acids in vitro. Synthetic cationic lipids designed to reduce the difficulties and dangers of liposome-mediated transfection can be used to prepare liposomes for in vivo transfection of genes encoding markers (Feigner et al., Proc. Natl. Acad) Sci. USA. 8 4:74 1 3 (1 9 8 7) ' Mackey et al., Proc. Natl. Acad. Sci. USA 8 5:8027 (1 98 8) and Ulmer et al., Science 25 9 :1 74 5 ( 1 993)). The use of cationic lipids enhances the encapsulation of negatively charged nucleic acids and also promotes fusion with negatively charged cell membranes (Feigner et al., Science 3 3 7:3 87 (1 9 8 9)). Lipid compounds and compositions which are particularly suitable for the transfer of nucleic acids are described in WO 09 5/1 8 86 3, W096/1 7823 and U.S. Patent No. 5,45,1,127. Transfection with liposomes to introduce exogenous groups -66 - 201207108 There are certain practical benefits to specific organs in vivo. Part of the advantage lies in the molecular targeting of liposomes to specific cells. It is clear that directing transfection of specific cell types will be particularly useful for tissues with cellular heterogeneity, such as pancreas, liver, kidney and brain. Lipids can be chemically coupled to other molecules for targeting purposes (Mackey et al. 1 988, supra). Targeted peptides (e.g., hormones or neurotransmitters) and proteins such as antibodies or non-like molecules can be chemically coupled to the liposomes. Other molecules can also be used to facilitate transfection of nucleic acids in vivo, such as cation oligopeptides (e.g., W095/21931), peptides derived from DNA binding proteins (e.g., WO 96/25508), or cationic polymers (e.g., W095/21931). It is also possible to introduce a carrier of naked DN A in vivo (see U.S. Patent Nos. 5,693,622, 5,5,89,466 and 5,580,85,9). Receptor-mediated DNA delivery methods can also be used (Curiel et al., Hum. Gene Ther. 3:1 47 (1 992) 3⁄4 Wu et al., J. Biol. Chem. 2 6 2:4429 (1 987) )) o The term "transfection" refers to the uptake of exogenous or heterologous RN A or DNA by cells. When exogenous or heterologous RNA or DNA is introduced into a cell, the cell is "transfected" by the RNA or DN A. When the transfected RN A or DNA causes a phenotypic change in the cell, the cell is "transformed" by exogenous or heterologous RNA or DNA. The transgeneic RNA or DNA can be integrated (covalently linked) to the chromosomal DN A of the genome of the constituent cells. "Transformation" refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in inheritance of the gene's stability. A host organism comprising the transmorphic nucleic acid fragment is referred to as a "gene transgenic" or "recombinant" or "transformed" organism. Further, a recombinant vector comprising a polynucleotide of the present invention may comprise one or more markers of replication, markers or selectable markers sought for amplification or expression in a cellular host. The term "selectable marker" refers to an identification factor, usually an antibiotic or chemical resistance gene, which can be selected based on the effect of the marker gene, ie resistance to antibiotics, resistance to herbicides, colorimetric markers , an enzyme, a fluorescent marker, and the like, wherein the effect is for tracking the inheritance of the nucleic acid of interest and/or identifying a cell or organism that has inherited the nucleic acid of interest. Examples of selectable marker genes known and used in the art include: providing amphetamine, streptomycin, jitamycin, kanamycin, hygromycin, bialaphos herbicide, sulfonamide, and a gene resistant to an analog; and a gene used as a phenotypic marker, namely an anthocyanin regulatory gene, a prenyltransferase gene, and the like. The term "reporter gene" refers to a nucleic acid encoding a recognition factor that can be identified based on the effect of the reporter gene, wherein the effect is used to track the inheritance of the nucleic acid of interest and to identify cells that have inherited the nucleic acid of interest. Or organisms, and/or measuring gene expression induction or transcription. Examples of reporter genes known and used in the art include: luciferase (Luc), green fluorescent protein (GFP), chloramphenicol acetyltransferase (CAT), beta-galactosidase (LacZ), beta - uridine bease (Gus) and the analog. A selectable marker gene can also be considered a reporter gene. "Promoter" and "promoter sequence" are used interchangeably and refer to a DN A sequence that controls the performance of a coding sequence or functional RN A. Generally, the coding sequence is located at the 3' end of the promoter sequence. Promoters may be derived in their entirety from the native -68-201207108 gene, or may be composed of different elements derived from different promoters found in nature, or even include synthetic DN A segments. Those skilled in the art understand that different promoters may direct genes to be expressed in different tissues or cell types, or at different stages of development, or in response to different environmental or physiological conditions. Promoters that cause genes to behave most of the time and in most cell types are often referred to as "constitutive promoters." Promoters that cause genes to behave in specific cell types are often referred to as "cell-specific promoters" or "tissue-specific promoters." Promoters that cause genes to behave during specific development or cell differentiation stages are often referred to as "development-specific promoters" or "cell differentiation-specific promoters." Promoters that are induced and cause gene expression after exposure or treatment with cells, biological molecules, chemicals, ligands, light, or analogs of inducible promoters are often referred to as "inducible promoters" or "regulatory" Sex promoter." It is also known that since in most cases the exact boundaries of the regulatory sequences are not fully defined, DNA fragments of different lengths may have the same promoter activity. In any of the vectors of the present invention, the vector optionally includes the promoter disclosed herein. In one embodiment, the promoter series is shown in the promoters of Table 1 herein. In any of the vectors of the present invention, the vector may optionally comprise a tissue-specific promoter. In one embodiment, the tissue-specific promoter is a tissue-specific promoter as disclosed herein. In another embodiment, the tissue-specific promoter series is shown in the tissue-specific promoter of Table 2 herein. The promoter sequence is usually bounded by the transcription start site of the 3' end, extending -69 - 201207108 to the upstream (5' direction) to include the minimum amount required to initiate transcription at a detectable amount above background 値. Base or component. Within the promoter sequence, a transcription initiation site (conveniently defined by, for example, the location of nuclease Sl), and a protein binding domain (consistent sequence) responsible for binding to RN A polymerase can be found. A "therapeutic switch promoter" ("TSP") is a promoter that controls the expression of a gene switch component. The gene switches and their various components are described in detail elsewhere in this specification. In certain embodiments, the TSP is constitutive, i.e., has continuous activity. A constitutive TSP may be fully constitutive (ie, generally functional in any tissue or cell without the need for additional factors or regulators) or tissue or cell-specific constitutive (ie, not required in a particular tissue type or cell type) Additional factors or regulators are generally functional). In certain embodiments, the TSP of the invention is activated under conditions associated with a disease, condition or condition. In certain embodiments of the invention involving two or more TSPs, the promoters can be a combination of constitutive and activating promoters. As used herein, "a promoter that is activated under conditions associated with a disease, condition, or condition" includes, but is not limited to, a disease-specific promoter, a promoter that responds to a particular physiological, developmental, differentiated, or pathological condition. A promoter that is responsive to a particular biological molecule, and a promoter specific for a particular disease, condition, or condition-related tissue or cell type (eg, tumor tissue or malignant cell). The TSP may comprise a sequence of a naturally occurring promoter, a modified sequence derived from a naturally occurring promoter' or a synthetic sequence (e.g., insertion of a response element into a minimal promoter sequence to alter the reactivity of the promoter). -70- 201207108 When RN A polymerase transcribes a coding sequence in a cell into mRN A, the coding sequence is under the control of transcriptional and translational control sequences, which are then trans-transformed by RNA (if the coding The sequence contains an intron and is translated into a protein encoded by the coding sequence. "Transcription and translation control sequences" refer to DN A regulatory sequences, such as promoters, enhancers, terminators, and the like, which are used to express coding sequences in a host cell. In eukaryotic cells, the polyadenylation signalling system controls the sequence.

The term "reactive element" refers to one or more cis-acting DNA elements that confer reactivity via a promoter that is mediated by interaction with the DNA-binding domain of the transcription factor. This DN A component may be palindrome (perfect or imperfect) in its sequence or consist of a sequence motif or a half site separated by an unequal number of nucleotides. The half sites may be similar or identical, arranged in a forward or reverse repeat, or as a single half site or a multimer of adjacent tandem sites. Depending on the nature of the cell or organism to be included in the response element, the reaction element may comprise a minimal promoter isolated from a different organism. In the presence or absence of a ligand, the DN A binding domain of the transcription factor binds to the DN A sequence of the response element to initiate or inhibit transcription of the downstream gene under the regulation of this response element. Examples of DNA sequences of response elements of the natural ecdysone receptor include: RRGG/TTCANTGAC/ACYY (SEQ ID NO: 25) (see Cherbas et. al., Genes Dev. 5: 120 (1991)); AGGTCAN(n) AGGTCA Wherein N(n) can be one or more spacer nucleotides (SEQ ID NO: 26) (see D'Avino et al., Mol. Cell. Endocrinol. 1 1 3:1 (1 995)); GGGTTGAATGAATTT (SEQ-71 - 201207108 ID NO: 27) (see Antoniewski et al., Mol. Cell Biol. 14: 4465 (1994)). The term "operably linked" means that the nucleic acid sequence is linked to a single nucleic acid fragment such that the function of one is affected by the other. For example, when a promoter can affect the performance of a coding sequence (i.e., the coding sequence is under the transcriptional control of a promoter), the promoter is operably linked to the coding sequence. The coding sequence may be operably linked in a synonymous or antisense orientation to the regulatory sequence. 用 The term "express" as used herein refers to the transcription and stable accumulation of synonymous (mRNA) or antisense RNA derived from a nucleic acid or polynucleotide. Performance can also refer to the translation of mRNA into a protein or polypeptide. The terms "card", "performance card" and "gene expression card" refer to a DN A segment that can be inserted into a nucleic acid or polynucleotide at a specific restriction site or by homologous recombination. The DN A segment comprises a polynucleotide encoding a polypeptide of interest, and the cassette and restriction sites are designed to ensure that the cassette is inserted into the appropriate reading frame for transcription and translation. "Transformed cassette" refers to a particular vector comprising a polynucleotide encoding a polypeptide of interest and, in addition to the polynucleotide, an element that facilitates transformation of a particular host cell. The cassette, the expression card, the gene expression cassette and the transposon of the present invention may also comprise elements which permit the polynucleotide encoding the polypeptide of interest to enhance expression in the host cell. Such elements include, but are not limited to, a promoter, a minimal promoter, an enhancer, a reaction element, a terminator sequence, a polyadenylation sequence, and the like. For the purposes of the present invention, the term "gene switch" refers to a combination of a promoter-related response element and a system based on a ligand-dependent transcription factor - 72-201207108, which is a ligand-dependent transcription factor. When the system of the substrate is present in one or more bodies, the regulatable gene appears to be involved in the reaction element and promoter. The phrase "polynucleotide encoding a gene switch" refers to a combination of a promoter-associated response element and a nucleotide encoding a ligand-dependent transcription factor-based system based on a ligand-dependent transcription factor. When one or more ligands are present, the regulatable gene appears to be included in the response element and the mimetic line. The therapeutic switch promoter of the invention can be any promoter useful for treating, preventing or preventing a particular disease, condition or condition. Examples include, but are limited to, promoters that exhibit increased expression only during a particular disease, condition, or condition and exhibit increased performance under specific cellular conditions (eg, proliferation, apoptosis, sputum changes, oxidative state changes, changes in oxygen levels). Promoter. In embodiments in which the gene switch comprises more than one transcription factor sequence, the specificity of the therapeutic method can be increased by combining a disease condition-specific promoter with a tissue or cell type-specific promoter, thereby limiting the therapeutic The organization in which the product is expressed. Thus, a panel or cell type specific promoter is included within the meaning of a therapeutic switch promoter. For example, a disease-specific promoter can be used to treat a promoter of a cancer promoter including an oncogene. Examples of classes of oncogenes include, but are not limited to, growth factors, growth factor receptors, protein kinases, planning cell death regulators, and transcription factors. Specific examples of oncogenes include, but are not limited to, sis, erb B, erb B-2, ras, abl, myc, and bc 1 -2 and TERT. Examples of his cancer-associated genes include tumor-associated antigen genes and other genes that are over-expressed in the tumor cells that are not due to PH-based or splicing in the tumor cells (eg, MAGE- 1. Carcinoembryonic antigen, tyrosinase, prostate specific antigen, prostate specific membrane antigen, p5 3 , MUC-1, MUC-2, MUC-4, HER-2/neu, T/Tn, MART-1 , gplOO, GM2, Τη, sTn and Thompson-Friedenreich antigen (TF)). Examples of promoter sequences and other regulatory elements (e.g., enhancers) that are known in the art and that can be used as therapeutic switch promoters of the invention are disclosed in the references listed in Tables 1 and 2 simultaneously. Promoter-associated diseases/disorders (representation or tissue specificity (Table 2) ° The promoter sequences disclosed in these references are hereby incorporated by reference in their entirety. The polynucleotides encoding any of the proteins listed in Table 1 are also The vector of the present invention having a promoter of a non-therapeutic promoter can be expressed. -74- 201207108 Table 1

Promoter Sequence Disease/Disease Patent/Public Application Her-2/neu(ERBB2/c-erbB-2) Cancer 5,518,885 Osteocalcin Calcified tumor 5,772,993 Matrix lysin-1 (stromelysin·1) Cancer 5,824,794 Prostate Specific antigen prostate cancer 5,919,652 human sodium-iodine co-transporter thyroid cancer 6,015,376 H19, IF-1, IGF-2 cancer 6,306,833 thymosin β15 breast cancer, pancreatic cancer, prostate cancer 6,489,463 Τ cytokine cancer 6,608,037 cartilage-derived retinoic acid sensitive Sexual protein chondrosarcoma, breast tumor 6,610,509 insulin pancreatic cancer 6,716,824 PEG-3 cancer 6,737,523 telomerase reverse transcriptase cancer 6,777,203 melanoma differentiation-related gene-7 cancer 6,841,362 prostate protein carcinoma 6,864,093 telomerase catalytic subunit; cyclin- Α cancer 6,936,595 midkine protein; c-erbB-2 carcinoma 7,030,099 prostate specific membrane antigen prostate cancer 7,037,647 p51 cancer 7,038,028 telomerase RNA cancer 7,084,267 prostate acid phosphatase prostate cancer 7,094,533 PCA3dd3 prostate cancer 7,138,235 DF3/MUC1 cancer 7,247,297 hex II cancer 2001/0011128 Cyclooxygenase-2 Cancer 2002/0107219 Super PSA Prostate Cancer 2003/0078224 skp2 Cancer 2003/0109481 PRL-3 Metastatic Colon Cancer 2004/0126785 CA125/M17S2 _ Nest Cancer 2004/0126824 IAI.3B Ovarian Cancer 2005/0031591 CRG -L2 Liver cancer 2005/0124068 TRPM4 Prostate cancer 2006/0188990 RTVP Glioma 2006/0216731 TARP Prostate cancer, breast cancer 2007/0032439 Telomere reverse transcriptase cancer 2007/0059287 A4 amyloid protein Alzheimer's disease 5,151,508 Amyloid β-protein precursor to Zimmer (Alzheimer's disease) 5,643,726 Alzheimer's disease Α4 amyloid protein pro-Merchheimer's disease 5,853,985 Neuropeptide FF CNS disease 6,320,038 -75- 201207108 Promoter sequence disease / Disease patent / open application endoplasmic reticulum pressure element pressure 7,049,132 urocortin π psychopathic disease 7,087,385 tyrosine hydroxylase neurological disease 7,195,910 complement factor 3 ; serum amyloid A3 inflammation 5,851,822 metalloproteinase 3 (TIMP-3 Tissue inhibitors rheumatism, cancer, autoimmune diseases, inflammation 5,854,019 p75 tumor necrosis factor receptor Immune disease 5,959,094 Tumor necrosis factor-α Inflammation 6,537,784 Peroxisome proliferator-activated receptor/ΠΑ-1 Non-pancreatic secretory phospholipaseΑ2 Inflammation 6,870,044 SOCS-3 Growth disease, autoimmune disease, inflammation 2002/0174448 SR- BI Lipid Disease 5,965,790 Ob Obesity 5,698,389 Site-1 Protease Obesity Diabetes 7,045,294 TIGR Glaucoma 7,138,511 VL30 Hypoxia 5,681,706 Excitatory Amino Acid Transporter-2 Nervous System Ischemia 2004/0171108 MDTS9 Renal Failure 2006/0014931 LIM, Pyrroline 5-carboxylate reductase, SIM2 Prostate disease 2006/0134688 Bax apoptosis 5,744,310 Fas apoptosis 5,888,764 bbc3 apoptosis 7,202,024 PINK-1 PI-3 kinase/Akt pathway disease 2006/0228776 -76- 201207108 Table 2 Start Subsequence Tissue Specific Patent/Public Application Troponin T Skeletal Muscle 5,266,488 myoD Muscle 5,352,595 Actin Muscle 5,374,544 Smooth Muscle Arterial Smooth Muscle 5,837,534 Anti-dystrophin-associated Protein Muscle 5,972,609 Myostatin Muscle 6,284,882 Smooth Muscle Myosin Heavy Chain Smooth Muscle6,780,610 cardiac angioprotein repeat protein myocardium 7,193,075 MLP muscle 2002/0042057 smooth muscle protein smooth muscle 2003/0157494 MYBPC3 cardiomyocyte 2004/0175699 Ταΐ α-tubulin neuron 5,661,032 intercellular adhesion molecule-4 (ICAM-4) neuron 5,753,502 γ- Aminobutyric acid type A receptor βΐ unit hippocampus back 6,066,726 neuron nicotinic acetylcholine receptor β2 subunit neurons 6,177,242 presenilin-1 neuron 6,255,473 calcium-calmodulin-dependent kinase ΙΙα forebrain 6,509,190 CRF2a receptor brain 7,071,323 nerve growth factor neurons 2003/159159 GLP-2 receptor intestine, brain 2002/0045173 type I transglutaminase keratinocytes 5,643,746 K14 keratinocytes 6,596,515 stearin coenzyme A desaturase skin 2002 510 510 510 510 510 510 Amyloid A breast duct epithelial cells 2005/010 7315 Endothelin-1 Endothelial Cell 5,288,846 -77- 201207108 Promoter Sequence Tissue Specific Patent/Open Application Serine Hematopoietic Cell 5,340,739 Platelet-Endothelial Cell Adhesion Molecule-1 (PECAM-l) Platelet, White Blood Cell, Endothelial Cell 5,668,012 Tie Receptor tyrosine kinase endothelial cells, bone marrow 5,877,020 KDR/flk-1 endothelial cells 5,888,765 Endoglin endothelial cells 6,103,527 CCR5 Bone marrow cells and lymphoid cells 6,383,746 CDlld Bone marrow cells 6,881,834 Platelet glycoprotein lib Hematopoietic cells 6,884,616 Pro-endothelin-1 Endothelial cells 7,067,649 Interleukin-18 binding protein monocytes 2006/0239984 CD34 Hematopoietic stem cells 5,556,954 Tec tyrosine kinase hematopoietic stem cells, liver 6,225,459 exhibit altered amounts of expression during a particular disease or condition and are therefore available for use in the present invention Other genes for the promoter include, but are not limited to, the genes listed in Table 3 (the relevant diseases/disorders are also listed). -78- 201207108

Table 3 Gene disease/disorder patent/public application MLH1, MSH2, MSH6, PMS1 'APC colorectal cancer 7,148,016 LEF-1 colon cancer 2002/0169300 F2 receptor colon cancer 2002/0187502 TGF-β type II receptor colon cancer 2004/0038284 EYA4 Colon cancer 2005/0003463 PCA3 Prostate cancer 7,138,235 K2 Prostate cancer 6,303,361 PROST 03 Prostate cancer metastasis 2002/0009455 PCAM-1 Prostate cancer 2002/0042062 PCADM-1 Prostate cancer 2003/0100033 PCA3dd3 Prostate cancer 2003/0165850 PCAV Prostate cancer 2006/0275747 PAcP Male Insensitivity Prostate Cancer 2006/0294615 SEQ ID NO: 1, Patent No. 5,866,329, incorporated herein by reference, PCT Serial No. PCT Serial No. </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt; SEQ ID NOS of 2007/0042420: 1 ' 3 Liver Cancer 2007/0042420 CSA-1 Chondrosarcoma 2001/0016649 SEQ ID NOS: 1 to 15 of U.S. Patent Application Publication No. 2001/0016651, which is incorporated herein by reference in its entirety, in Here, pancreatic cancer 2003/0212264 SYG972 Breast cancer 2002/0055107 Urb-ctf Breast cancer 2003/0143546 -79- 201207108 Gene disease/disorder patent/public application BCU399 Breast cancer 2003/0180728 TBX2 Breast cancer 2004/0029185 Cyr61 Breast cancer 2004/0086504 DIAPH3 Breast cancer SEQ ID NOs: 1 to 24 of U.S. Patent Application Publication No. 2007/0134669, incorporated herein by reference in its entirety, in, in, in, in, in in in in in in in in in in in in in in in in in in in in in in in in in in /0102263 ΒΕΗΑΒ CNS Cancer 2003/0068661 IL-8 Kaposi's sarcoma 2003/0096781 SEQ ID NO: 1 to 278 of U.S. Patent Application Publication No. 2002/0198362, hereby incorporated herein incorporated by reference in its entirety, in B cell carcinoma 2003/0147887 BP1 leukemia 2003/0171273 DAP-kinase, HOXA9 non-small cell lung cancer 2003/0224509 ARP Renal clear cell carcinoma, inflammatory disease 2004/0010119 Nbk Kidney Cancer 2005/0053931 CD43 Ovarian Cancer 2006/0216231 US Patent Application Publication No. 2007/0054268 SEQ ID NO: 1 to 84, incorporated herein by reference, ovarian cancer 2007/0054268 P7-hcG &gt; βό-hCG ' p6e -hCG ' p5-hCG &gt; βδ-hcG ' p3-hCG uterine tumor 2006/0292567 MTAls hormone insensitive cancer 2006/0204957 Old-35, Old-64 Tumor hyperplasia 2003/0099660 LAGE-1 Cancer 6,794,131 CIF150/hTAF „ 150 cancer 6,174,679 P65 tumor-fetoprotein cancer 5,773,215 -80- 201207108 Gene disease/disorder patent/public application telomerase cancer 2002/0025518 CYP1B1 cancer 2002/0052013 14-3-3σ cancer 2002/0102245 NES1 cancer 2002/0106367 CAR- 1 Cancer 2002/0119541 HMGI' MAG Cancer 2002/0120120 ELL2 Cancer 2002/0132329 Ephrin B2 Cancer 2002/0136726 WAF1 Cancer 2002/0142442 CIF130 Cancer 2002/0143154 C35 Cancer 2002/0155447 BMP2 Cancer 2002/0159986 BUB3 Cancer 2002/0160403 Polymerase K Cancer 2003/0017573 EAG1 'EAG2 Cancer 2003/0040476 SEQ ID NO: 18, 20, 22 of US Patent Application Publication No. 2003/0044813, incorporated herein by reference herein. 3/0051260 HLTF Cancer 2003/0082526 Barx2 Cancer 2003/0087243 SEQ ID NO: 18, 20, 22, 32, 34, 36 of US Patent Application Publication No. 2003/0108920, incorporated herein by reference herein. 2003/0109443 Pp 32rl Cancer 2003/0129631 BMP4 Cancer 2003/0134790 TS10q23.3 Cancer 2003/0139324 Nuclear spindle-associated protein carcinoma 2003/0157072 PFTAIRE Cancer 2003/0166217 SEMA3B Cancer 2003/0166557 -81 - 201207108 Gene disease/disorder patent/ Published application MOGp cancer, multiple sclerosis, inflammatory disease 2003/0166898 Fortilin cancer 2003/0172388 US Patent Application Publication No. 2003/0215833 SEQ ID NO: 1, incorporated herein by reference to cancer 2003/0215833 IGFBP-3 Cancer 2004/0005294 Polyhomeotic 2 Cancer 2004/0006210 PNQALRE Cancer 2004/0077009 SEQ ID NO: :1, 3 of US Patent Application Publication No. 2004/0086916, incorporated herein by reference herein. Cancer 2004/0146877 miR15, miR16 Cancer 2004/0152112 Head and neck serine protease inhibitor (Headpin) Cancer 2004/0180371 PAOhl/SMO Cancer 2004/0229241 Hippo, Mst2 Cancer 2005/0053592 PSMA-like cancer, neurological disease 2005/0064504 JAB1 cancer 2005/0069918 NF-AT cancer 2005/0079496 P28ING5 cancer 2005/0097626 MTG16 cancer 2005/0107313 ErbB-2 cancer 2005/0123538 HDAC9 cancer 2005/ 0130146 GPBP Cancer 2005/0130227 MG20 Cancer 2005/0153352 KLF6 Cancer 2005/0181374 ARTS1 Cancer 2005/0266443 Dock 3 Cancer 2006/0041111 Annexin 8 Cancer 2006/0052320 MH15 Cancer 2006/0068411 -82- 201207108 Gene disease/disorder patent/ Published application DELTA-N p73 Cancer 2006/0088825 RapR6 Cancer 2006/099676 StarDIO Cancer 2006/0148032 Cizl Cancer 2006/0155113 HLJ1 Cancer 2006/0194235 RapR7 Cancer 2006/0240021 A34 Cancer 2006/0292154 Sef Cancer 2006/0293240 Killin Cancer 2007/ 0072218 SGA-1M carcinoma 2007/0128593 TGF-β type II receptor cancer 2002/0064786 GCA-related gene giant cell arteritis 6,743,903 PRV-l true erythrocytosis 6,686,153 SEQ ID NO: 2, 4, US 5,948,637 Incorporate here by reference. 5,948,637 Vezfl Vascular Disease 2002/0023277 MLP Dilated Cardiomyopathy 2002/0042057 VEGI Disease Rational angiogenesis 2002/0111325 PR0256 Cardiovascular disease 2002/0123091 AOP2 Atherosclerosis 2002/0142417 Remodeling protein arterial restenosis, fibrosis 2002/0161211 Phosphodiesterase 4D Stroke 2003/0054531 Prostaglandin receptor subtype EP3 Periphery Arterial obstructive disease 2003/0157599 CARP Heart disease 2004/0014706 HOP Congenital heart disease 2004/0029158 SEQ ID NO: 1 to 4 of US Patent Application Publication No. 2004/0087784, hereby incorporated by reference herein by reference. 0087784 -83- 201207108 Gene disease/disorder patent/public application PLTP Atherosclerosis, vascular disease, hypercholesterolemia, Tangier's disease, familial HDL-deficient disease 2006/0252787 US Patent Application Publication SEQ ID NO: 1, 3 to 8, 15, 16 ' of 2007/0160996 is incorporated herein by reference. Thrombosis 2007/0160996 UCP-2 Stroke 2002/0172958 FLJ11011 Fanconi Anemia 2006/0070134 Codanin -1 Anemia 2006/0154331 SEQ ID NO: 1, 6, 8 ' of US 5, 763, 591 is incorporated herein by reference for insulin dependence Urinary disease, 5,763,591 resistin type 2 diabetes 2002/0161210 Archipelin Diabetes 2003/0202976 SEQ ID NO: 2, 7, 16, 27 of US Patent Application Publication No. 2004/0053397, incorporated herein by reference, for diabetes, hyperlipidemia Disease 2004/0053397 Neuronatin Metabolic Disorder 2004/0259777 Ncb5or Diabetes 2005/0031605 7B2 Endocrine Disease 2005/0086709 PTHrP ' ΡΕΧ Metabolic bone disease 2005/0113303 KChIPl Type 2 diabetes 2005/0196784 SLIT-3 Type 2 diabetes 2006/ 0141462 CX3CR1 Type 2 Diabetes 2006/0160076 SMAP-2 Diabetes 2006/0210974 SEQ ID NO: 2, 8, 12, 16, 22, 26, 28, 32 of US Patent Application Publication No. 2006/0228706, incorporated herein by reference Type 2 diabetes 2006/0228706 IC-RFX Diabetes 2006/0264611 E2IG4 Diabetes, insulin resistance, obesity 2007/0036787 -84- 201207108

SEQ ID NO: 2, 8, 10, 14, 18, 24, 26, 30, 34, 38, 44, 50, 54, 60, SEQ ID NO: US Patent Application Publication No. 2007/0122802 62, 68, 74, 80, 86, 92, 98, 104, 110, incorporated herein by reference. Diabetes 2007/0122802 UCP2 Body Weight Disease 2002/0127600 Ob Receptor Body Weight Disease 2002/0182676 Ob Weight Disease 2004/0214214 Dpi Nerve Degenerative disorders 2001/0021771 NRG-1 Schizophrenia 2002/0045577 Synaptic protein III Schizophrenia 2002/0064811 NRG1AG1 Schizophrenia 2002/0094954 AL-2 Neuronal disorders 2002/0142444 Proline dehydrogenase bipolar disorder , severe depression, schizophrenia, obsessive-compulsive disorder 2002/0193581 MNR2 Chronic neurodegenerative diseases 2002/0197678 ATM Ataxia microvascular dilatation syndrome 2004/0029198 Ho-1 Dementia 2004/0033563 CON202 Schizophrenia 2004/0091928 Ataxin-1 Neurodegenerative disorders 2004/0177388 NR3B Exercise neuron disorders 2005/0153287 NIPA-1 Hereditary spastic paraplegia 2005/0164228 DEPP, adrenomedullin, csdA schizophrenia Disease 2005/0227233 Inf-20 Neurodegenerative Disease 2006/0079675 EOPA Brain Development and Degenerative Diseases 2007/0031830 SERT White Closure 2007/0037194 FRP-1 Glaucoma 2002/0049177 -85- 201207108 Gene Disease/Disease Patent/Public Application Serum Amyloid A Glaucoma 2005/0153927 BMP2 Osteoporosis 2002/0072066 BMPR1A Juvenile Polyposis 2003/0072758 ACLP Abdominal Disruption 2003/0084464 Resistin-like Molecule β Familial Adenomatous Caries, Diabetes, Insulin Resistance, Colon Cancer, inflammatory bowel disease 2003/0138826 Dlg5 Inflammatory bowel disease 2006/0100132 SEQ ID NO: 1 to 82 of US Patent Application Publication No. 2002/0119452, incorporated herein by reference to osteoarthritis 2002/0119452 TRANCE immune system disorder 2003/0185820 Matrix protein-3 Osteoarthritis 2003/0203380 Synovial protein rheumatoid arthritis 2004/0152871 SEQ ID NO: 9, 35 of US Patent Application Publication No. 2007/0028314, incorporated herein by reference. 2007/0028314 HIV LTR HIV infection 5,627,023 SHIVA HIV infection 2004/0197770 EBI 1 ΈΒΙ 2 ' EBI 3 EB disease Infection 2002/0040133 NM23 Family Skin &quot;J, Intestinal Disorder 2002/0034741 US Patent Application Publication No. 2002/0169127 SEQHD NO: 1, incorporated herein by reference Psoriasis 2002/0169127 Eps8 Skin disorders, wound healing 2003/0180302 3-10 Thyroid Pathology 2002/0015981 SEQ ID NO: 2 of US Patent Application Publication No. 2003/0207403, incorporated herein by reference to thyroid condition 2003/0207403 -86- 201207108 Gene disease/disorder patent/public application US patent SEQ ID NO: 3 of Application Publication No. 2007/0020275, incorporated herein by reference, thyroid disorder 2007/0020275 Hair follicle growth factor alopecia 2003/0036174 Keratin-chain astrocyte alopecia 2003/0211065 GCR9 Asthma, lymphoma, leukemia 2003/0166150 US Patent SEQ ID NOS: 1 to 71 of the application publication No. 2004/0002084, incorporated herein by reference in its entirety in the s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s ID NO: 1 to 16, incorporated herein by reference. Endometriosis 2002/0127555 FGF23 Hypophosphatemic disorder 2005/0156014 BBSR Bardet Biedl's syndrome 2003/0152963 MIC-1 Fetal abnormalities, cancer, inflammatory diseases, miscarriage, premature labor 2004/0053325 MIA-2 Liver injury 2004/0076965 IL-17B cartilage degenerative disease 2004/0171109 Hyperthyroidism producing enzyme Polysulfate lipase deficiency 2004/0229250 LPLA2 Alveolar proteinosis 2006/0008455 CXCL10 Respiratory disease 2006/0040329 US Patent Application Publication No. 2006/0140945 SEQ ID NO: 1, 2, incorporated herein by reference to kidney disease 2006/0140945 HFE2A Iron Metabolic Disease 2007/0166711 -87- 201207108 Once a gene that is regulated in a disease, condition or condition is identified, The promoter of this gene can be used in the gene switch of the present invention. Sequences of many genes, including promoter regions, are known in the art and can be obtained from public databases such as the GenBank gene database. Thus, once the appropriate gene is identified, the promoter sequence can be readily identified and obtained. Another aspect of the invention pertains to identifying a suitable gene for which the promoter of the promoter can be isolated for placement in a gene switch. Thus, the nature of the gene is not relevant to particular embodiments of the invention, as long as the promoter can be isolated for subsequent settings or environments. The invention thus encompasses the use of a promoter from a gene that is not recognized by ticks. Once the appropriate gene is identified, the genetic sequence required to determine promoter function is routine or experimental. In fact, a variety of commercial procedures are available to assist in the determination of the promoter region of a gene of interest. For example, Ding et al. recently elucidated the promoter sequence of the novel Sprouty4 gene by gradually deleting the 5'-flanking sequence of the human Sprouty4 gene (Am. j. Physiol. Lung Cell. Mol. Physiol. 287: L5 2 (2004) &gt; incorporated herein by reference). Briefly, when the transcription start site is determined, a PCR fragment is generated using a common PCR primer to select a segment of the 5'-flanking segment in a single direction. The resulting segment was cloned into a luciferase receptor vector and the activity of luciferase was measured to determine the promoter region of the human Spr〇uty4 gene. Another example of a procedure for obtaining and validating a gene promoter includes the steps of: (1) obtaining disease and non-disease cell/tissue samples of similar/identical tissue types; and (2) isolating all RN A or mRN A from the samples; 3) Perform differential microarray analysis of disease and non-disease RNA; (4) identify candidate disease-specific transcripts; (5) find genomic sequences related to disease-specific transcripts; -88 - 201207108 (6) Or synthesizing DNA sequences upstream and downstream of the predicted transcription initiation site of the disease-specific transcript; (7) designing and producing a promoter reporter vector using DN A of different lengths from step 6; and (8) Testing promoters in diseased and non-disease cells/tissues and unrelated cells/tissues The source of the promoter into which the promoter is inserted into the gene switch can be either natural or synthetic, and the source of the promoter should not be limited here. The scope of the invention is described. In other words, the promoter may be directly colonized from the cell, or the promoter may be selected from a different source, or the promoter may be synthetic. Gene Switching System Gene switching may be any gene switch that regulates gene expression by adding or removing specific ligands. In one embodiment, the gene is associated with a gene switch in which the amount of gene expression depends on the amount of ligand present. Examples of ligand-dependent transcription factor complexes useful in the gene switch of the present invention include, but are not limited to, individual ligands thereof (e.g., glucocorticoids, estrogens, luteinizing hormones, retinoids, ecdysone, and the like) A member of the nuclear receptor superfamily activated by the analog or mimetic) and rTTA activated by tetracycline. In one aspect of the invention, the gene is associated with an EcR-substrate gene switch. Examples of such systems include, but are not limited to, U.S. Patent No. 6,258,603, U.S. Patent No. 7,045,315, U.S. Patent Application Serial No. 2006/00 1 47 1 1 , 2007/0 1 6 1 086 and International Publications The system described in WO 0 1/70 8 1 6 . Examples of chimeric ecdysone receptor systems are described in U.S. Patent No. 7,091,03, U.S. Patent Publication No. -89-201207108

Application Nos. 2002/0110861, 2004/0033600, 2004/0096942, 2005/0266457 and 2006/0 100416 and International Publication No. WO 0 1 /708 1 6 , WO 02/0666 1 2 , WO 02/0666 1 3, WO

02/0666 1 4 'WO 02/06 66 1 5 WO 02/2907 5 and WO 2005/1 0 86 1 7 , each of which is incorporated herein by reference in its entirety. An example of a non-sterol acute peel agonist modulating system is the Rheo Switch® mammalian inducible expression system (New England Biolabs, Ipswich, Iowe). In another aspect of the invention, the gene-opening relationship is based on heterodimerization of FK506-binding protein (FKBP) with FKBP rapamycin-associated protein (FRAP) and is composed of rapamycin or Regulation of immunosuppressive analogs. Examples of such systems include, but are not limited to, recording technology (ARIAD Pharmaceuticals, Cambridge, MA) and US 6,0, 5, 7 09, 6, 1 1, 7, 0 0, 6,479, 65 3, 6, 1 8 The system described in the patents 7,75 and 6,649,595. In one embodiment, the gene switch comprises a single transcription factor sequence encoding a ligand-dependent transcription factor complex under the control of a therapeutic switch promoter. The transcription factor sequence may encode a naturally occurring or artificial ligand-dependent transcription factor complex. An artificial transcription factor is a transcription factor in which the native sequence of the transcription factor has been altered, e.g., by sequence mutation or by combining domains from different transcription factors. In one embodiment, the transcription factor comprises a purine nuclear receptor ligand binding domain. In one embodiment, the scorpion nuclear receptor ligand binding domain is derived from a receptor for the ecdysone receptor, a ubiquitous receptor (UR), an orphan receptor 1 (OR-1), a steroid hormone nuclear receptor 1 ( NER-l), retinoid X receptor interaction protein-15 (RIP-15), liver-90-201207108 X receptor P (LXRp), steroid hormone receptor-like protein (RLD-1), liver X receptor Body (LXR), liver X receptor a (LXRa), farnesoid X receptor (FXR), receptor interacting protein 14 (RIP-14) or farnesol receptor (HRR-1). In another embodiment, the sputum nuclear receptor LBD is derived from the ecdysone receptor.蜕. ecdysone base gene switch

EcR and other sputum groups are members of the nuclear receptor superfamily of the system, wherein all members are generally characterized by an amine group optionally fused to a heterodimeric partner (HP) to form a coactivator protein (CAP). a trans-acting domain (AD, also referred to interchangeably as "TA" or "TD"), a DNA binding domain (DBD), and an LBD fused to a DBD via a hinge region to form a ligand-dependent transcription factor (LTF) ). The term "DNA binding domain" as used herein includes a DNA binding protein from a minimal polypeptide sequence to a full length DNA binding protein' as long as the function of the DNA binding domain is linked to a specific reaction element. Members of the nuclear receptor superfamily also include four or five domains: A/B έ , , C, D, Ε, and some members have F domains (see US Patent No. 4,981,784 and Evans, Science 240) :889( 1 98 8)). The "Α/Β" domain corresponds to the transactivation domain, "C" corresponds to the DNA binding domain, "D" corresponds to the hinge region, and "E" corresponds to the ligand binding domain. Some family members also have another transactivation domain on the carboxy-terminal side of the LPD corresponding to "F". The following polypeptide sequences are reported as the polypeptide sequence of the ecdysone receptor (peeling steroid receptor) (20-hydroxy-ecdysone receptor) (2 〇E receptor) (EcRH) (the composition of the nuclear receptor subfamily 1) Member 1), Genbank's registration number is P3402 1. -91 - 201207108 The cutaneous hormone receptor (878 amino acids) from Drosophila melanogaster (SEQ ID NO: 5) 1 mkrrwsnngg fmrlpeesss evtsssnglv lpsgvnmsps sldshdycdq dlwlcgnesg 61 sfggsnghgl sqqqqsvitl amhgcsstlp aqttiiping nangnggstn gqyvpgatnl 121 galangmlng gfngmqqqiq nghglinstt pstpttplhl qqnlggaggg giggmgilhh 181 angtpnglig vvgggggvgl gvggggvggl gmqhtprsds vnsissgrdd lspssslngy 241 sanescdakk skkgpaprvq eelclvcgdr asgyhynalt cegckgffrr svtksavycc 301 kfgracemdin ymrrkcqecr lkkclavgmr pecvvpenqc amkrrekkaq kekdkmttsp 361 ssqhggngsl asgggqdfvk keildlmtce ppqhatipll pdeilakcqa rnipsltynq 421 laviykliwy qdgyeqpsee dlrrimsqpd enesqtdvsf rhiteitilt vqlivefakg 481 lpaftkipqe dqitllkacs sevmmlrmar rydhssdsif fannrsytrd sykmagmadn 541 iedllhfcrq mfsmkvdnve yalltaivif Sdrpglekaq Iveaiqsyyi dtlriyilnr 601 hcgdsmslvf yakllsilte lrtlgnqnae mcfslklknr klpkfleeiw dvhaippsvq 661 shlqitqeen erleraermr asvggaitag idcdsastsa aaaaaqhqpq pqpqpqpssl 721 tqndsqhqtq pqlqpqlppq lqgqlqpqlq pqlqt Qlqpq iqpqpqllpv sapvpasvta 781 pgslsavsts seymggsaai gpitpattss itaavtasst tsavpmgngv gvgvgvggnv 841 smyanaqtara almgvalhsh qeqliggvav ksehstta DBD is characterized by two zinc cysteine fingers, in which two amino acid phantom P boxes and D boxes are given, the two phantoms give a reaction Component specificity. These domains may be chimeras of natural domains, modified domains or different domains of heterologous receptor proteins. EcR, like a subgroup of the nuclear receptor family, also has a region that is less well defined and that causes heterodimerization. Since the domains of nuclear receptors are modular in nature, LBD, DBD, and AD may be interchangeable. In another embodiment, the transcription factor comprises AD, DBD, and a sputum nuclear receptor LBD'. The DBD recognizes a response element linked to a therapeutic protein or therapeutic polynucleotide to modulate the therapeutic protein or therapeutic multinuclear The performance of glycosides. In certain embodiments, the purine nuclear receptor L B D comprises a substitutional mutation. In another embodiment, the gene switch comprises a first transcription factor sequence, such as CAP, under the control of a first therapeutic switch promoter (TSP-1), and a second therapeutic switch promoter (TSP · 2) control a second transcription factor sequence, such as L·TF, wherein the protein encoded by the first transcription factor sequence 歹u and the second transcription factor-92-201207108 subsequence interact to form a protein complex (LDTFC), ie "Double switch j or "hybrid" is the base gene switch. The first and second TSPs may be the same or different. In this embodiment, the presence of two different TSPs in the gene switch required to represent the therapeutic molecule enhances the specificity of the therapeutic approach (see Figure 2). Figure 2 also shows the ability to modify a therapeutic gene switch to treat any disease, condition or condition by simply inserting the appropriate TSP. In another embodiment, the first and second transcription factor sequences, e.g., CAP or LTF, are under the control of a single therapeutic switch promoter (e.g., TSP-1 in Figure 1). Activation of this promoter will produce CAP and LTF in a single open reading frame. This can be achieved by using a transcriptional linker such as an IRES (internal ribosome entry site). In this embodiment, the two portions of the ligand-dependent transcription factor complex are synthesized upon activation of TSP-1. TSP-1 may be a constitutive promoter or only activated under conditions associated with a disease, condition or condition. In another embodiment, a transcription factor sequence, such as LTF, is under the control of a therapeutic switch promoter, The promoter is only activated under conditions associated with the disease, condition or condition (eg, TSP-2 or TSP-3 in Figure 4), and another transcription factor sequence such as CAP is at the constitutive therapeutic switch promoter (eg, Under the control of TSP-1). In this embodiment, a portion of the ligand-dependent transcription factor complex is constitutively present, however the second portion will be synthesized only under conditions associated with the disease, condition or condition. In another embodiment, a transcription factor sequence such as CAP is under the control of a -TSP (eg, TSP-1 in Figure 3), and two or more different -93-201207108 second transcription factor sequences such as LTF The -1 and LTF-2 are under the control of different TSPs (e.g., TSP-2 and TSP-3 in Figure 3). In this embodiment, various LTFs may have different DBDs that recognize different factor regulatory promoter sequences, such as DBD-A and Factor Regulatory Promoter-l (FRP-l)-related response elements, DBD-B and Factor Regulatory Promoter-2 (FRP-2)-related response element binding. Various factor regulatory promoters can be operably linked to different therapeutic genes. In this way, multiple treatments can be provided simultaneously. In one embodiment, the first transcription factor sequence encodes a polypeptide comprising AD, DBD, and a sputum nuclear receptor LBD, the DBD identifying a response element linked to the therapeutic product sequence to modulate the performance of the therapeutic product sequence, The second transcription factor sequence encodes a transcription factor comprising a nuclear receptor LBD selected from the group consisting of a vertebrate retinoid X receptor (RXR), an invertebrate RXR, an extra-valve protein (USP) or comprising at least two different A chimeric nuclear receptor for a nuclear receptor ligand binding domain polypeptide fragment of a vertebrate RXR, an invertebrate RXR or USP (see WO 01/708 1 6 Α 2 and US 2004/0096942 Α 1). The "concomitant" nuclear receptor ligand binding domain may additionally comprise a truncation mutation, a deletion mutation, a substitution mutation or other modification. In another embodiment, the gene switch comprises a first transcription factor sequence encoding a first polypeptide and a second transcription factor sequence encoding a second polypeptide comprising a nuclear receptor LBD and a DBD, the DBD Identifying a response element linked to a therapeutic product sequence to modulate the performance of the therapeutic product sequence'. The second polypeptide comprises AD and a nuclear receptor LBD, wherein one of the nuclear receptor LBD is a nuclear receptor LBD. In one embodiment, the first polypeptide is substantially free of AD and the second polypeptide is substantially free of DBD. For the purposes of the present invention -94-201207108, "substantially free" means that the problematic protein does not contain sufficient sequence of the problematic domain to provide activation or binding activity. In another aspect of the invention, the first transcription factor sequence encodes a protein comprising a heterodimer with AD ("CAP"). The second transcription factor sequence encodes a protein comprising DBD and LBD ("LTF" ). When there is only one nuclear receptor LBD sputum group LBD, the other nuclear receptor LBD may be derived from any other nuclear receptor to form a dimer with the sputum group LBD. For example, when the nucleus receptor LBD is EcR LBD, other nuclear receptor LBD partners may come from EcR, vertebrate RXR, invertebrate RXR, super-valve protein (USP) or contain at least two different A chimeric nuclear receptor selected from the group consisting of a vertebrate RXR, an invertebrate RXR or a USP nuclear receptor LBD polypeptide fragment (see WO 0 1 /708 1 6 A2, International Patent Application PCT/US02/05 23 5 and US) 2004/0096942 A1, which is incorporated herein in its entirety by reference. The "concomitant" nuclear receptor ligand binding domain may additionally comprise truncation mutations, deletion mutations, substitution mutations or other modifications. In one embodiment, the vertebrate RXR LBD is from Homo sapiens

U (Homo sapiens), mouse musculus, Rattus norvegicus, Gallus gallus, Sus scrofa domestica, xenopus laevis, RXR of the zebrafish zebrafish (Danio rerio), the tunicate, the Polyandrocarpa misakiensis or the Trippedalia cysophora.

In one embodiment, the 'invertebrate RXR ligand binding domain is derived from the super-valve polypeptide of Locusta migratoria ("LmUSP-95-201207108"), and the Amblyomma americanum. RXR homolog 1 ("AmaRXRl"), RXR homolog 2 ("AmaRXR2") of C. sinensis, and RXR homolog of Cecala pugilator ("CpRXR") ), RXR homolog of the beetle Tenebrio molitor ("TmRXR"), RXR homolog of the bee mellifera ("AmRXR"), RXR of Myzus persicae RXR homolog of a homolog ("MpRXR") or a non-dipterous/non-Lepidopteran insect. In one embodiment, the chimeric RXR LBD comprises at least two RXR homologs selected from the group consisting of a vertebrate species RXR polypeptide fragment, a non-vertebrate species RXR polypeptide fragment, or a non-dipterous insect/non-Lepidopteran insect invertebrate species. A polypeptide fragment of a polypeptide fragment. A chimeric RXR ligand binding domain for use in the invention may comprise RXR polypeptide fragments of at least two different species, or when the species are identical, the two or more polypeptide fragments may be derived from two or more of the RXR polypeptide fragments of the species Different isomers. Such chimeric RXR LBD lines are disclosed, for example, in WO 2002/066614. In one embodiment, the chimeric RXR ligand binding domain comprises at least one vertebrate species RXR polypeptide fragment and an invertebrate species RXR polypeptide fragment. In another embodiment, the chimeric RXR ligand binding domain comprises at least one vertebrate species RXR polypeptide fragment and a non-dipteran insect/non-lepidary insect invertebrate species RXR homolog polypeptide fragment. When the ligand is combined with the LB D of the nuclear receptor and in turn binds to the response element of the therapeutic product sequence-associated FRP, the ligand provides an expression of the external temporal regulation -96-201207108 therapeutic product sequence. The combination of the various components of the invention, i.e., binding of the ligand to the LBD, binding of the DBD to the reactive element, binding of the AD to the promoter, etc., is not critical. In a particular embodiment, the ligand binds to the LBD of the sputum nuclear receptor and to the nuclear receptor LBD to allow expression of the therapeutic product sequence. This machine does not eliminate the possibility that the ligand binds to the sputum nuclear receptor (GHNR) or its partner and results in the formation of an active homodimeric complex (eg GHNR + GHNR or companion + partner). Preferably, the hybrid gene switch is produced in one or more different receptor domains. Typically, one or more of the three domains DBD, LBD, and AD can be selected from a source other than the source of the other domain such that the hybrid gene and the formed hybrid protein are reversed in the selected host cell or organism. The activation activity, complementary binding to the ligand, and identification of specific reactive elements are optimized. Furthermore, the reaction element itself may be modified or substituted by other DN Α binding protein domain reaction elements, such as the GAL-4 protein from yeast (see Sadowski et al., Nature 335: 563 (1988)) or from the large intestine. LexA protein of Bacillus (see Brent et al., Cell 43:729 (1 985)), or synthetic response elements specifically designed to interact with proteins designed, modified, and selected for specific interactions (See, for example, Kim et al., Proc. Natl. Acad. Sci. USA, 94:361 6 (1997)) to accommodate hybrid receptors. Another benefit of the two-hybrid system is that they are able to select promoters for driving gene expression based on the desired end result. These dual controls are particularly important in the field of gene therapy, especially when cytotoxic proteins are produced, because the cells that exhibit the timing and performance can be controlled. When a gene operably linked to an appropriate promoter is introduced into a cell of an individual, the expression of the exogenous gene is controlled by the presence of the system of the present invention. Promoters may be constitutively or inducibly regulated, or may be tissue specific (i.e., expressed only in a particular cell type) or specific for certain stages of development of an organism. In the presence or absence of a ligand, the DNA binding domain of the first hybrid protein binds to the DN A sequence of the response element to initiate or inhibit transcription of the downstream gene under the regulation of the response element. Functional LDTFCs, such as EcR complexes, may also contain additional proteins such as immunophilins. Additional members of the nuclear receptor family of proteins known as transcription factors (such as DHR38 or PFTZ-1) may also be ligand-dependent or non-ligand-dependent partners of Ecr, USP and/or RXR. In addition, other cofactors may be required, such as proteins commonly referred to as coactivators (also known as aptamers or mediators). These proteins do not specifically bind to the DN A sequence and do not participate in basal transcription. They may demonstrate their effects on transcriptional activation via a variety of mechanisms, including stimulation of DN A binding to activators, affecting chromatin structure, or mediator activator initiation complex interactions. Examples of such coactivators include RIP140, TIF1, RAP46/Bag-1, ARA70, SRC-1/NCoA-1, TIF2/GRIP/NCoA-2, ACTR/AIBl/RAC3/pCIP, and hybrid coactivator C reactions. Element B binds to the protein CBP/p300 (see review literature Glass et al., Curr. Opin. Cell Biol. 9:222 (1997)). In addition, protein cofactors commonly referred to as co-repressors (also known as inhibitors, silencers or silent mediators) may be required to effectively inhibit transcriptional activation in the absence of a ligand. These co-repressors can interact with the ligand-free E c R to silence the activity of the reaction element. Current evidence suggests that -98-201207108, ligand binding alters the conformation of the receptor, resulting in the release of co-repressors and the attraction of the above-mentioned co-activators, thereby eliminating their silent activity. Examples of co-repressors include N-CoR and SMRT (see review literature Horwitz et al., Mol Endocrinol. l〇: 1167 (1996)). These cofactors may be endogenous factors in the cell or organism, or may be exogenously added in a regulated or unregulated manner. B. Rapamycin Substrate Gene Switch The present invention further provides a gene switch system using FK506 binding protein as a ligand-dependent transcription factor complex and rapamycin as a ligand. In one embodiment, the construct of the coding gene switch comprises (a) a first polynucleotide encoding a first chimeric protein, the first polynucleotide and rapamycin or an analog thereof Binding and comprising at least one FK506-binding protein (FKBP) domain and at least one protein domain heterologous to the FKBP domain, wherein the FKBP domain comprises a peptide sequence selected from the group consisting of: (1) naturally occurring FKBP (2) a naturally occurring FKBP variant in which up to 10 amino acid residues are deleted, inserted or substituted with a substituted amino acid' or (3) by FKBP with (1) or (2) a FKBP encoded by a DNA sequence that selectively hybridizes to a DNA sequence; (b) a second polynucleotide encoding a second chimeric protein, the second polynucleotide and U) rapamycin or rapamycin The analog and (b) the first chimeric protein form a complex and comprise at least one FKBP: rapamycin-99-201207108 binding (FRB) domain and at least one protein domain heterologous to the FRB domain, Wherein the FRB domain comprises a peptide sequence selected from the group consisting of: (4) a naturally occurring FRB structure (5) a variant of a naturally occurring FRB domain in which up to 1 amino acid residue is deleted, inserted or substituted with a substituted amino acid, or (6) is encoded by (4) or (5) The FRB domain encoded by the DNA sequence of the FRB DNA sequence selectively hybridizes. In this gene switch system, the first polynucleotide and the second polynucleotide are each under the control of one or more therapeutic switch promoters described elsewhere herein. In addition, in certain embodiments, at least one protein domain heterologous to the FKBP and/or FRB domains of the first and second chimeric proteins may be one or more "active" or "effect" Domain. The effector domain can be selected from a wide variety of protein domains, including DNA binding domains, transcriptional activation domains, cell localization domains, and signaling domains (ie, when clustered or multimerized, can cause cell growth, proliferation, Domains of differentiation, apoptosis, gene transcription, etc.). In certain embodiments, a fusion protein comprises at least one DNA binding domain (eg, a GAL4 or ZFHD1 DNA binding domain) and another fusion protein comprises at least one transcriptional activation domain (eg, a VP 16 or P65 transcriptional activation domain) ). Ligand-mediated ligation of the fusion protein represents the formation of a transcription factor complex and a target gene that results in the initiation of transcription and DNA sequence ligation (i.e., binding) by the DNA binding domain of one of the fusion proteins. Information about gene expression systems and ligands is disclosed in the United States on the 6th, 1 8 7, 7 5 7 B 1, 6, 6 4 9, 5 9 5 B 1, 6, 5 0 9, 1 5 2 B 1 -100- 201207108 6,479,653 B1 and 6,117,680 B1 patents. In other embodiments, the invention provides a gene switch system comprising a polynucleotide encoding two fusion proteins that self-aggregate in the absence of a ligand, wherein (a) the first fusion protein comprises conditions a sexual aggregation domain 'this domain binds to a selected ligand and a transcriptional activation domain' and (b) the second fusion protein comprises a conditional aggregation domain that binds to a selected ligand and DN A Domain binding, and (c) in the absence of a ligand, the cells exhibit a gene operably linked to a regulatory DN A that binds to the DN A binding domain. The modified cells comprising the gene switch system are expanded in the presence of a ligand sufficient to inhibit the amount of the gene. Ligand removal induces the expression of encoded proteins that cause cell death. The nucleic acid encoding the two fusion proteins is under the control of at least one conditional promoter. A gene expression system utilizing a conditional aggregation domain is disclosed in U.S. Patent Publication No. 2002/0048792. C. Prokaryotic Suppressor/Operator Subgen Gene Switching System In one embodiment, the invention provides a gene switching system comprising (a) a first polynucleotide encoding a transactivation fusion protein, the trans The activation fusion protein comprises a prokaryotic tetracycline ("t et") inhibitor and a eukaryotic transcriptional activator protein domain; and (b) a second polynucleotide encoding a therapeutic protein or therapeutic polypeptide, wherein the second The polynucleotide line is operably linked to a minimal promoter and at least one tet operator sequence. The first polynucleotide encoding a transactivator fusion protein can comprise a therapeutic switch promoter as described elsewhere herein. The expression of the lethal protein is up-regulated in the absence of tetracycline - 101 - 201207108 (see for example 0〇536116131. (1992)?|'〇(:.&amp;11· Acad. Sci. 89: 5547-5 55 1 ' Gossen et al. (1 993) TIBS 18 : 471 -475 ' Furth et al. (1 994) Proc. Natl. Acad. Sci. 91: 9302-9306 and Shockett et al. (1 995) Proc. Natl. Acad. Sci. 92: 6522-6526). The TetO expression system is disclosed in U.S. Patent No. 5,464,758 B. In another embodiment, the gene switch system comprises a lactose ("Lac") inhibitor from Escherichia coli. - an operating subsystem. The gene switch system of the present invention may further comprise (a) a first polynucleotide encoding a transactivation fusion protein comprising a prokaryotic lac I repressor and a eukaryotic transcriptional activator protein Domain: and (b) a second polynucleotide encoding a therapeutic protein or therapeutic polypeptide wherein the second polynucleotide is operably linked to a therapeutic switch promoter. In a Lac system, In the presence of lactose-free or synthetic analogues such as isopropyl-bD-thiogalactoside, the iac operon is not Other gene switch systems include those described in the following: US Patent No. 7,091,038, WO2004078924, EP1266015, US200 1 0044 1 5 1 , US200201 1 0861, US20020 1 1 952 1 , US2004003 3 600 , US20040197861, US2004023 5 097 , US20060020 1 46 , US20 04 004943 7 , US20040096942 , US20050228016 , US20050266457 , US20060100416 , W0200 1 /7081 6 , W02002/29075 , W02002/066612 , W02002/0666 1 3 , W02002/0666 1 4 , W02002/0666 1 5, W02005/1 086 1 7 , US Patent No. 6,258,603 '-102-201207108 US2005 0209283, US200502280 1 6 , US20060020 1 46 , EP096 5644, US Patent No. 7, 3 0 4, 1 2 2, US 7th Patent No. 3 0 4,1 6 1 , MX234742, KR 1 0-05 63 1 43 , AU765 3 06, AU2002-248500 and AU2002-3 065 50 ° D. Combination of Gene Switching Systems The present invention provides two or more a nucleic acid composition of a gene switch system, a modified cell, and a bioreactor, wherein the gene switch system comprises an activation of one or more ligands in an effective amount a ligand-dependent transcription factor complex, wherein the two or more gene switch systems comprise a first gene switch and a second gene switch, each selectively inducing one or more therapeutic properties when combined with one or more ligands The performance of a polypeptide or therapeutic polynucleotide. The scope of the invention encompasses any number and/or combination of gene switch systems. In one embodiment, the invention provides a nucleic acid composition comprising: a. a first gene switch system comprising: i. a first gene expression cassette, the cassette comprising a polynucleotide encoding a first hybrid polymorphism An acid, the first hybrid polypeptide comprising: 1. a transactivation domain that activates a factor regulatory promoter operably linked to a polynucleotide encoding a therapeutic polypeptide or a therapeutic polynucleotide; and 2. a second partner gene, wherein the second gene exhibits a cassette, the cassette comprises a polynucleotide encoding a second hybrid polymorphism, the second hybrid polypeptide comprising: l. DNA binding domain, which recognizes and encodes therapeutic a polypeptide or a -103-201207108 therapeutic polynucleotide polynucleotide operably linked to a factor regulatory promoter; and 2. a ligand binding domain; and iii. a third gene expression cassette, the cassette encoding A therapeutic poly- or polynucleotide of a therapeutic polynucleotide comprising: 1. a factor-regulated promoter activated by a trans-activation domain of a second hybrid polypeptide; and 2 - encoding a therapeutic polypeptide Therapeutic multinucleus An acid polynucleotide, and b. a second gene expression system comprising: i. a first gene expression cassette comprising a polynucleotide encoding a first hybrid polypeptide, the first hybrid polypeptide comprising: a transactivation domain that activates a factor regulatory promoter operably linked to a polynucleotide encoding a therapeutic polypeptide or therapeutic polynucleotide; and 2 a heterodimeric partner domain, ii. a second gene A cassette comprising a polynucleotide encoding a second hybrid polypeptide comprising: a DNA binding domain that recognizes and encodes a polynucleotide encoding a therapeutic polypeptide or a therapeutic polynucleotide An operably linked factor regulatory promoter: and 2 a ligand binding domain; and iii. a third gene exhibiting a cassette comprising a polynucleotide encoding a therapeutic polypeptide or a therapeutic polynucleotide, the multinuclear Glycosidic acid comprises: -104- 201207108 1. A factor regulatory promoter activated by a transactivation domain of a second hybrid polypeptide; and 2. a polynucleotide encoding a therapeutic polypeptide or a therapeutic polynucleotide An inducible gene expression system provides that a given therapeutic polynucleotide or therapeutic polypeptide behaves under conditions associated with different diseases, conditions or conditions, or under the same conditions associated with the same disease, condition or condition, or A variety of therapeutic polypeptides or therapeutic polynucleotides are present under different conditions associated with a disease, condition or condition. In certain embodiments, the combination of the two or more gene switch systems can be (1) a dual-switched cutaneous hormone receptor basal gene expression system, and (2) a single-switched cutaneous hormone receptor base gene switch. In other embodiments, the combination can be (1) a single or dual switch ecdysone receptor base gene switch and (2) a rapamycin base gene switch. Alternatively, the combination of gene switch systems can be two identical rapamycin substrate gene switch systems as disclosed above. Combinations of any of the possible gene switch systems are within the scope of the invention. Examples of dual-switched acute serotonin systems can be found, for example, in WO 2002/29075 and US 2002/0110861 ° Ligand The term "ligand" as used herein, when applied to LDTTC-substrate gene switches, such as EcD complex substrate gene switches, is described A small and soluble molecule that activates a gene switch to stimulate the expression of the polypeptide encoded therein. A ligand for a ligand-dependent transcription factor complex of the present invention and a protein comprising one or more ligand-105-201207108 binding domain, a heteromeric partner domain, a DNA binding domain, and a transactivation domain Complex binding. The choice of ligand for the activating ligand-dependent transcription factor complex is determined by the type of gene switch utilized. Examples of ligands include, but are not limited to, dermatological steroids such as ecdysone, 20-hydroxy quercetin, cedarone a, midazolam a and the analogs, 9-cis retinoic acid, retinoic acid synthesis Sexual analogs, sputum, sulphonium, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate, sulphate U.S. Patent Application 2,04/0 1 7 1 65 1 describes oxadiazolines; benzhydrylalkylcyanoguanidines, such as those disclosed in European Application No. 46 1,809; -NW-二芳醯基讲's such as those disclosed in U.S. Patent No. 5,225,443; Ν-醯-Ν-院----- carbonyl hydrazine, such as disclosed in European Application No. 23 4,994: Ν-芳醯基-Ν- Alkyl-Ν'-aryl fluorenyl hydrazines, such as those described in U.S. Patent No. 4,985,,,,,,,,,,,,,,,,,; Similar materials include 3,5-di-tertiary butyl-4-hydroxy-indole-isobutyl-benzamide, 8-0-ethylh-palapa, oxysterol, 22(R) hydroxyl Cholesterol, 24(S)hydroxycholesterol, 25-epoxycholesterol, Τ〇9〇1317, 5_α-6-α-epoxycholesterol-3-sulfate (ECHS), 7-ketocholesterol-3-sulfate , farnesol, bile acid, 1, bisphosphonate, juvenile hormone III and the like. Examples of the dimercaptopurine ligand which can be used in the present invention include RG-115819 (3,5-dimethyl-benzoic acid N-(l-ethyl-2,2-dimethyl-propyl)-N' -(2-methyl-3-methoxy-benzimidyl)-oxime), RG-115932((R)-3,5-dimethyl-benzoic acid N-(l-tertiary butyl) - -106- 201207108 Butyl)-N'-(2-ethyl-3-methoxy-benzhydryl)_醯肼) and RG_115830 (3,5-dimethyl-benzoic acid^(1) _Tri-tert-butyl-butyl)_;^|_(2-ethyl-3-methoxy-benzimidoxime). See, for example, U.S. Patent Application Serial No. 12/155,111 and ? :1 [Application? (: 171; 32008/006757, both cases are incorporated herein by reference. For example, the ligand for the ecdysone receptor basal gene switch may be selected from any suitable ligand. Naturally occurring Ecdysone or ecdysone analogue (eg 20-hydroxyepiticin, midazolidone a, ponasterone A, ponasterone B, ponasterone C, 26-iodosinone A, achyranthes Or both 26-methylsulfonium oxacillinone and non-sterol inducers can be used as ligands for the gene switch of the present invention. US Patent No. 6,379,945 B1 An insect steroid receptor ("HEcr") isolated from Heliothis virescens. This receptor acts as a gene switch that responds to steroids and certain non-steroid inducers. In this system and many other pairs In systems where both steroids and non-sterol inducers are reactive, nonsteroidal inducers have significant advantages over steroids, including, for example, lower manufacturing costs, metabolic stability, absence from insects, plants. Or mammalian and environmentally acceptable. U.S. Patent No. 6,379,94 5 B1 describes the use of two kinds of benzoyl hydrazines, namely 1,2-diphenylmethyl decyl-1-tributyl-anthracene and fenfenol. (tebufen〇Zide)(N-(4-ethylbenzhydryl)·Ν'-(3,5-dimethylbenzylidene)-Ν'-tertiary butyl-hydrazine) as a ecdysone base The ligand of the gene switch. The ligand of the present invention also includes other benzoyl hydrazines, such as those disclosed in U.S. Patent No. 5,1,7,7,7,1, using tebufenozide as Chemical ligand of the cutaneous hormone receptor of Drosophila melanogaster -107-201207108 U.S. Patent No. 6,147,282. In addition, a non-limiting example of a dermatological ligand is 3,5-di-tert-butyl-4-hydroxyisobutyl-benzamide, 8-0-acetamidine. Chihapaxan, 1,2-diylhydrazine, N, substituted-N,N, disubstituted anthracene, diphenylmercaptoalkylcyanoguanidine, N-substituted-N-alkyl-N,N- Diaryl fluorenyl, N-substituted-N-fluorenyl-N-alkylcarbonyl hydrazine or N-aryl fluorenyl-hydrazine'-alkyl-hydrazone--indenyl hydrazine (see U.S. Patent No. 6,723,531). In one embodiment, the ligand for the ecdysone base gene switch system is a dimercaptopurine ligand or a chiral dimercaptopurine ligand. The ligand for the gene switch system can be a compound of formula I

Wherein the fluorenyl alkoxy group, the arylalkyloxy group or the aryloxy group; the B group may be an optionally substituted aryl group or an optionally substituted heteroaryl group; and each of R1 and R2 may be optionally substituted An alkyl group, an arylalkyl group, a hydroxy group, a halogenated alkyl group, an optionally substituted cycloalkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted heterocyclic group, An aryl group which may be optionally substituted or a heteroaryl group which may be optionally substituted; or a pharmaceutically acceptable salt, hydrate, crystalline form or amorphous form thereof. In another embodiment, the ligand may be a compound of the formula 11 enriched in the image -108 - 201207108

Formula II

a heteroaryl group substituted with a aryloxy group or an arylalkyl group; an optionally substituted heteroaryl group A, an aryl group which may be optionally substituted, or may be optionally substituted with an B group. Any optionally substituted aryl or R1 and R2 are each independently substituted alkyl, arylalkyl, hydroxyalkyl, halogenated, substituted, optionally substituted ring, which may be optionally substituted a radical, an optionally substituted alkynyl group, an optionally substituted heterocyclic group, an optionally substituted aryl group or an optionally substituted heteroaryl group; provided that R1 is not equal to R2; wherein R1 and R2 are carried The absolute configuration of a symmetric carbon atom is predominantly s-type; or a pharmaceutically acceptable salt, hydrate, crystalline form or amorphous form. In certain embodiments, the ligand may be a mirror-isomerized compound of formula III

109- 201207108 A is a oxy group, an aryl-based oxy group, an aryloxy group, an aryl group, an optionally substituted aryl group or an optionally substituted heteroaryl group; the B system may be optionally substituted An aryl group or an optionally substituted heteroaryl group; and each of R1 and R2 is an optionally substituted alkyl group, an arylalkyl group, a hydroxy group, a halogenated alkyl group, an optionally substituted ring-based group, or An optionally substituted alkenyl group, an optionally substituted block group, an optionally substituted heterocyclic group, an optionally substituted aryl group or an optionally substituted heteroaryl group; provided that R1 is not equal to R2; And the absolute configuration of the asymmetric carbon atom of R2 is predominantly r-type; or a pharmaceutically acceptable salt, hydrate, crystalline form or amorphous form. In one embodiment, the ligand may be at least 95% of (R)-3,5-dimethyl-benzoic acid N-(l-tertiary butyl-butyl)-N'-( 2-Ethyl-3-methoxy-benzimidyl)-oxime, or a pharmaceutically acceptable salt, hydrate, crystalline form or amorphous form thereof. The dimercaptopurine ligand of the formula I and the chiral dimercaptopurine ligand of the formula II or III are used in the ecdysone base gene switch system to provide a therapeutic polypeptide or therapeutic polynucleoside of the invention from an external temporal state The way the acid is expressed. See U.S. Patent Application Serial No. 12/155, filed on May 29, 2008, which is hereby incorporated by reference. The ligand used in the present invention can form a salt. The term "salt" as used herein means an acidic and/or basic salt formed with an inorganic and/or organic acid and a base. In addition, when the compound of formula I, II or III contains both a basic group and an acidic group, a zwitterion ("internal salt") may be formed, which is still included in the term "salt" as used herein. "." Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are used, for example, in the separation or purification steps employed in the preparation, although other salts may also be employed. The salt of the compound of the formula I, II or III can be formed, for example, by reacting the compound with an amount such as an equivalent amount of an acid or a base, which is carried out in a medium in which the salt can be precipitated or in an aqueous medium, followed by freezing and drying. .配 A ligand containing a basic group can form a salt with various organic and inorganic acids. Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acids such as trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenes. Sulfonate, hydrogen sulfate, borate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanoate, digluconate, lauryl sulfate, ethanesulfonic acid Salt, fumarate, glucoheptonate, glycerol phosphate, hemisulfate, heptanoate, hexanoate, hydrochloride (formed with hydrochloric acid), hydrobromide (with hydrogen bromide) Forming), hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate (formed with maleic acid), methanesulfonate (formed with methanesulfonic acid), 2-naphthalene Sulfonate, nicotinic acid salt, nitrate, oxalate, jelly, peroxosulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, water Salicylate, succinate, sulfate (such as those formed with sulfuric acid), sulfonates (such as those mentioned herein), tartrate, thiocyanate , Such as a tosylate tosylate (t〇sylate), - and salts of such analogs. -111 - 201207108 Ligands containing acidic groups can form salts with various organic and inorganic salts. Exemplary basic salts include ammonium salts, alkali metal salts such as sodium salts, lithium salts and phosphonium salts, alkaline earth metal salts such as calcium salts and magnesium salts, salts containing organic bases such as organic amines such as benzathine, and Cyclohexylamine, heimbamin (formed with N,N-bis(dehydrodecanoyl)ethylenediamine), methyl-D-glucosamine, N-methyl-D-glucosamine, tertiary butyl Base amines and salts containing amino acids such as arginine, lysine and the like. A non-limiting example of a ligand for an inducible gene expression system utilizing the FK506 binding domain is FK5 06, cyclosporin A or rapamycin. FK500, rapamycin and their analogs are disclosed in U.S. Patent Nos. 6,649,595 B2 and 6,1,8,7,7,7. See also US Patent Nos. 7,276,498 and 7,273,874. The ligands described herein can be administered separately or as part of a pharmaceutical composition comprising a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is in the form of a solution, suspension, lozenge, capsule, ointment, elixir or injectable composition. In one embodiment, the vectors and methods of the invention can be used to represent polynucleotides encoding proteins comprising, but not limited to, interleukins, immunomodulators, coagulation factors, antibodies or antibody fragments, tumor necrosis factor Receptors (TNFR) such as etanercept, erythropoietin, α-l antitrypsin, interferon (IFN), interferon alpha, interferon beta, interferon gamma, interferon-beta-la, Interferon-β-lb, seventh factor, factor VIII, ninth factor, antithrombin III, hepatitis B virus protein, hormones such as growth hormone (GH), human growth hormone (hGH), parathyroid hormone ( PH), -112- 201207108 Thyroid stimulating hormone (TSH), GCSF or a fragment thereof, GM-CSF or a fragment thereof. In one embodiment, the polynucleotide encoding the antibody encodes a monoclonal antibody 〇 In another embodiment, the vectors and methods of the invention can be used to express nucleic acids as a vaccine. The invention also provides a vaccine composition comprising a vector or expression system of the invention. In another embodiment, the vaccine composition comprises an adjuvant. In the case of a gene switch, the term "ecdysone receptor substrate" refers to a gene switch comprising at least one functional portion of a naturally occurring or synthetic sarcokinin receptor ligand binding domain, when the ligand is associated with an ecdysone receptor. This gene switch regulates gene expression when the ligand binding domain binds. Examples of the ecdysone responsiveness system are described in U.S. Patent Nos. 7,091,03 and 6,25,603. In one embodiment, the system is a RheoSwitch® therapeutic system (RTS) comprising two fusion proteins, a DEF of a mutated ecdysone receptor (EcR) fused to a Q-domain of a Gal4 DNA binding junction. The domain and the EF domain of the chimeric RXR fused to the VP16 transcriptional activation domain are shown under the control of a constitutive promoter as shown in Figure 1. The term "modulating" refers to the induction, reduction or inhibition of nucleic acid or gene expression, which results in the induction, reduction or inhibition of the production of a protein or polypeptide, respectively. The polynucleotide or vector of the present invention may further comprise at least one promoter suitable for driving the expression of the gene in a host cell. Enhancers useful in embodiments of the invention include, but are not limited to: SV40 enhancer, cytomegalovirus (CMV) enhancer, elongation factor 1 (EF1) -113-201207108 enhancer, yeast enhancer, viral gene Enhancers and analogs. Termination control regions (i.e., terminators or polyadenylation sequences) can also be derived from a variety of genes that are native to the preferred host. Optionally, the termination site is not necessary but preferably included. In one embodiment of the invention, the termination control region may comprise or be derived from a synthetic sequence, a synthetic polyadenylation signal, a SV40 late polyadenylation signal, a SV40 polyadenylation signal, bovine growth hormone ( BGH) polyadenylation signal, viral termination sequence or the analog. The term "3 'non-coding sequence" or "3 'non-translated region (UTR)" refers to a DNA sequence located downstream (3') of a coding sequence, which may comprise a polyadenylation [poly(A)] recognition sequence and other encodings. The signal is modulated to affect the sequence of mRNA processing or gene expression. The polyadenylation signal is typically characterized by affecting the addition of a polyadenylation sequence to the 3' end of the mRNA precursor. "Regulatory region" refers to a nucleic acid sequence that modulates the expression of a second nucleic acid sequence. The regulatory regions may include sequences that are inherently responsible for the expression of a particular nucleic acid (homologous region), or sequences that are of different origin but are responsible for representing different proteins or even synthetic proteins (heterologous regions). In particular, the sequences may be sequences or derived sequences of prokaryotic, eukaryotic or viral genes that stimulate or inhibit transcription of the gene in a specific or non-specific manner and in an inducible or non-inducible manner. The regulatory region includes an origin of replication, an RN A cleavage site, a promoter, an enhancer, a transcription termination sequence, and a signal sequence that directs the polypeptide into the secretory pathway of the target cell. The regulatory region of the "heterologous source" refers to A regulatory region in which the expressed nucleic acid is naturally associated. The heterologous regulatory regions comprise regulatory regions from different species -114-201207108, regulatory regions from different genes, hybrid regulatory sequences, and regulatory sequences that are not found in nature but are designed by those of ordinary skill in the art. "RNA transcript" refers to a product produced by rnA polymerase catalyzed transcription of a DNA sequence. When the RNA transcript DNA sequence is perfectly complementary, it is preferably a primary transcript or it may be an RNA sequence derived from post-transcriptional processing of the primary transcript, referred to as mature RNA. "Messenger RNA (mRNA)" refers to RNA that has no introns and can be translated into proteins by cells. "Γ" refers to a double-stranded DNA complementary to mRNA and derived from the mRNA. "Synonymous RNA" refers to an RNA transcript that includes mRNA and thus can be translated into a protein by a cell. "Antisense RNA" refers to an RNA transcript that is complementary to all or part of the target primary transcript or mRNA and blocks the expression of the target gene. Antisense RNA may be complementary to any portion of a particular gene transcript, such as a 5&apos; non-coding sequence, a 3&apos; non-coding sequence or a coding sequence. "Functional RNA" refers to antisense RNA, ribozyme RNA, or other RNA that is not translated but has an effect on cellular processes. "Polypeptide", "peptide" and "protein" are used interchangeably and refer to a polymeric compound consisting of a covalently linked amino acid residue. "Isolated polypeptide", "isolated peptide" or "isolated protein" means substantially free of such compounds normally associated with it in its natural state (eg, other proteins or polypeptides, nucleic acids, carbon water) a polypeptide or protein of a compound, a lipid). "Isolated" does not mean the exclusion of artificial or synthetic mixtures with other compounds, or the presence of impurities which do not affect biological activity, which may be due, for example, to incomplete purification, addition of stabilizers or chemical synthesis into pharmaceutically acceptable formulations. -115- 201207108 "Substitution of a mutant polypeptide" or "substitution mutation" will be understood to mean a mutant polypeptide comprising at least one Sr-type or naturally occurring amino acid substitution, which is associated with a wild-type or naturally occurring polypeptide. Amino acids are different. Substitutional Mutants Polypeptides may contain only one wild-type or naturally occurring amino acid substitution, referred to as a "point mutation" or a "single point mutation" polypeptide. Alternatively, the substitution mutant polypeptide may comprise one or more wild-type or naturally occurring amino acids substituted with two or more amino acids different from the wild-type or naturally occurring polypeptide. According to the present invention, a purine-receiving receptor ligand-binding domain polypeptide comprising a substitution mutation comprises at least one wild type substituted with an amino acid different from the wild-type or naturally occurring purine nuclear receptor ligand-binding domain polypeptide. Or naturally occurring amino acids. When a substituted mutant polypeptide comprises a substitution of two or more wild-type or naturally occurring amino acids, the substitution may comprise an equal number of wild-type or naturally occurring amino acid deletions for substitution, ie 2 wild-type or naturally occurring The amino acid is substituted by 2 non-wild-type or non-naturally occurring amino acids, or the substitution may comprise an unequal amount of wild-type amino acid deletion for substitution, ie 2 wild-type amino acids are 1 The non-wild type amino acid substitution (substitution + deletion mutation) or the two wild type amino acids are substituted by three non-wild type amino acids (substitution + insertion mutation). Substitution mutations can be described using the abbreviated nomenclature system to indicate the amino acid residues and numbers substituted in the reference polypeptide sequence and the substituted amino acid residues. For example, a substitution mutation in which the 20th amino acid residue of a polypeptide is substituted may be abbreviated as "Χ20ζ", wherein "X" is a substituted amino acid '"2〇" is the amino acid The position or number of the residue in the polypeptide, and the new amino acid substituted by the oxime. Therefore, interchangeably abbreviated as "Ε 2 〇 A" or "-116- 201207108

The substitution mutation of Glu2 0Ala" indicates that the mutation comprises glutamic acid (commonly abbreviated as "E" or "Glu") substituted at the polypeptide position 20 with an alanine residue (usually abbreviated as "A" or "Ala"). Substitution mutations can be performed using any of the mutation-forming techniques known in the art, including but not limited to in-situ site-directed mutagenesis (Hutchinson et al., J. Biol. Chem. 253:6551 (1978); Zoller et al., DN A 3 :479 (1 984); Oliphant et al., Gene 4 4:1 77 (1 986); Hutchinson et al., Pro c. Natl. Acad. Sci. USA 83:710 (1986)), using TAB® connection Subgen (Phar mac ia), restriction endonuclease digestion/fragment deletion and substitution, PCR vector/oligonucleotide-directed mutagenesis and the like. PCR-based techniques are more suitable for site-directed mutagenesis (see Higuchi, 1 989, &quot;Using PCR to Engineer DN A&quot;, in PCR Technology: Principles and Applications for DN A Amplification, H. Erlich, ed., Stockton Press, Chapter 6, pages 61 to 70). The term &quot;fragment&quot; as applied to a polypeptide refers to a polypeptide having a shorter amino acid sequence than the amino acid sequence of the reference polypeptide, and the polypeptide comprises identical amino acid sequences compared to all portions of these reference polypeptides. When appropriate, the fragments may be included within a larger polypeptide to form part of a larger polypeptide. The fragments of the polypeptide of the invention may be at least 2, 3, 4, 5, 6, 8, 10, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22' 25, 26 30, 35, 40, 45, 50, 100, 200, 240 ^ 300 or more amino acids. A "variant" of a polypeptide or protein refers to any segment, derivative or mutation derived from a polypeptide or protein and which retains at least one biological property of the polymorph or protein. Different polypeptides or proteins are mutated in nature. These variants may be allelic variants characterized by different nucleotide sequences encoding the protein, or may be involved in post-translational modifications. A skilled person can produce variants that have single or multiple generations, deletions, additions or substitutions. These variants are: (a) variants in which one or more amino acid residues are replaced by a conservative or carboxylic acid, (b) one or more amino acid peptides or variants of the protein, a variant of one or more amine groups' and (d) a variant in which the polypeptide or protein is fused to another serum albumin. Obtaining the properties of these variants (inhibition, deletion of 'mutations, etc.), chemistry and enzymes It is known to those skilled in the art. In one embodiment, the variant is less than about 14 amino acids. The term "homology" refers to the percentage of two polynucleotides or two multiple identity. From a group The sequence and another sequence can be determined by techniques known in the art, for example, by directly comparing the sequence information between two polypeptide molecules to the sequence information and using an easy to use computer program by homology. The conditions for the formation of stable double-strand between the regions are increased, and then the size is determined by single-strand specific nuclease digestion to determine homology. All grammatical forms and spelling differences used herein refer to play The relationship between the proteins of the "source", the flakes may have a qualitative structural group, and the differential cleavage of the heavy amino acid may include the addition of a particularly non-conservative amine to the polyacid including substitution of a polypeptide such as a technique including The homology of the polypeptides in the field to the columns between the peptide groups can be deduced, for example, by the arrangement. Alternatively, the phrase "hybridization" of the fragments which can be hybridized by nucleotides, includes from -118 to 201207108 Proteins of superfamilies (such as the immunoglobulin superfamily) and homologous proteins from different species (such as myosin light chain, etc.) (Reeck et al., Cell 5 0: 667 (1 98 7)). (and their coding genes) have sequence homology because they have a high degree of sequence similarity. However, in general usage and in this application, when the term "homologous" is modified by an adverb such as "highly" The term "homologous" may refer to sequence similarity rather than a source of co-evolution. Therefore, the term "sequence similarity" in all grammatical forms refers to nucleic acids that may or may not share proteins of co-evolving origin. The degree of identity or correspondence between amino acid sequences (see Reeck et al., Cell 50: 667 (1 987). In one embodiment, at least about 50 of the defined length of the two DNA sequences. Nucleotide matching of % (eg, at least about 75 %, 90%, or 95 %), which are "substantially homologous" or "substantially similar." Substantially homologous sequences can utilize sequences Standard software comparison sequences available in the database are identified or identified by Southern hybridization experiments, for example, under stringent conditions as defined by a particular system. Defining appropriate hybridization conditions is a matter of skill in the art (see, for example, Sambrook et al., 1989, supra.. "Substantially similar" as used herein refers to a change in one or more nucleotide bases that results in the substitution of one or more amino acids, but does not affect the protein encoded by the DNA sequence. A nucleic acid fragment of functional properties. "Substantially similar" also refers to a nucleic acid fragment in which alteration of one or more nucleotide bases does not affect the ability of the nucleic acid fragment to alter the expression of the vector by antisense or co-suppression techniques. "Substantially similar" also refers to modifications of the nucleic acid fragments of the invention, such as deletion or insertion of one or more nucleotide bases that do not materially affect the functional properties of the formed transcript. It will therefore be appreciated that the present invention encompasses not only this particular -119-201207108 exemplary sequence. Modification of each of the proposals and determination of whether the encoded product retains biological activity are within the skill of the art. In addition, those skilled in the art will recognize that substantially similar sequences encompassed by the present invention may also be cleaned by them under stringent conditions (0.1 times SSC, 0.1% SDS, 65 C, and 2 times SSC, 0. 1% SDS and then The ability to hybridize with the sequence exemplified herein is defined by 0.1 times SSC '0.1% SDS wash. Substantially similar nucleic acid fragments of the invention are at least about 70%, 80 °/ of the DN A sequences of the DN A sequences and the nucleic acid fragments reported herein. , 90% or 95% - a fragment of the resulting nucleic acid. When the two amino acid sequences have more than about 40% of the amino acid, or more than 60% of the amino acids are similar (functionally identical), the two amino acid sequences are "substantially homologous" or "substantially" similar". Preferably, the similar or homologous sequence is arranged by using, for example, GCG (Genetics Computer Group, GCG Suite Program Manual, 7th Edition, Madison, Wisconsin). Compare it to forensics. The term "corresponding" as used herein refers to a sequence of similar or homologous sequences, whether or not the exact position is the same or different from the molecule that measures similarity or homology. The nucleic acid or amino acid sequence alignment may include a spacer. Thus, the term "corresponding" refers to sequence similarity, rather than the numbering of amino acid residues or nucleotide bases. The "significant portion" of the amino acid or nucleotide sequence contains sufficient amino acid sequence of the polypeptide or the nucleotide sequence of the gene, which can be manually evaluated by those skilled in the art, or by using algorithms such as BLAST. (Basic partial alignment search tool; Altschul et al., J. Mol. Biol. -120- 201207108 2 1 5:403 (1 993), available from ncbi.nlm.nih.gov/BLAST/) for computer automated sequence comparison And identifying to identify the polypeptide or gene by presump. Generally, a sequence of 10 or more than 10 contiguous amino acids or 30 or more than 30 nucleotides is required to presuppose the homology of a polypeptide or nucleic acid sequence to a known protein or gene. In addition, for nucleotide sequences, gene-specific oligonucleotide probes containing 20 to 30 contiguous nucleotides can be used for gene identification (eg, southern hybridization) and isolation (eg, bacterial colonies or phage plaques). Sequence dependent method for in situ 〇 hybridization). Further, a short oligonucleotide having 12 to 15 bases can be used as an amplification primer in PCR to obtain a specific nucleic acid fragment containing the primer. Thus, a "significant portion" of a nucleotide sequence comprises sufficient sequence to specifically recognize and/or isolate a nucleic acid fragment comprising the sequence. The term "percent identity" as used in the art refers to two or more polymorphic sequences or two. The relationship between one or more polynucleotide sequences is determined by comparing the sequences. In this context, "consistency" also refers to the degree of sequence correlation between a polypeptide or a polynucleotide sequence, and may be determined by the matching between such sequences, as appropriate. "Consistency" and "similarity" can be easily calculated by known methods, including but not limited to the following description: Computational Molecular B i ο 1 〇gy (L esk, AM, ed.) Oxford University Press, New York ( 1988); Biocomputing: Informatics and Genome Proj ects (Smith, DW, ed.) Academic Press, New York (1 993); Computer Analysis of Sequence Data, Part I (Griffin, AM, and Griffin, HG, eds.) Humana Press, New Jersey ( 1 994) ; Sequence Analysis in Molecular -121 - 201207108

Biology(v〇n H e in je, G., e d.) A c ad emic Press ( 1 9 8 7); and Sequence Analysis Primer (Gribskov, M. and Devereux, J., eds.) Stockton Press , New York (1991). A preferred method of determining identity is designed to give the best match between test sequences. Methods for determining consistency and similarity are compiled in publicly available computer programs. Sequence alignment and percent consistency calculations can be performed using sequence analysis software, such as the Megalign program of the Lasergene Bioinformatics computer suite (DNASTAR, Madison, Wisconsin). The multiple alignment of the sequence can be performed using the Clustal alignment method and preset parameters (notch penalty = 1 〇, gap length penalty = 10) (Higgins et al., CABIOS. 5: 151 (1989)). Preset parameters for the two-to-two ratio using the Clustal method can be selected from: KTUPLE 1, GAP PENALTY = 3, WINDOW = 5, and DIAGONALS SAVED = 5. The term "sequence analysis software" refers to any computer algorithm or software program that can be used to analyze nucleotide or amino acid sequences. "Sequence Analysis Software" is commercially available or self-developed. Typical sequence analysis software includes, but is not limited to, GCG suites (Wisconsin Suite version 9.0, Genetics Computer Group (GCG), Madison, Wisconsin), BLASTP, BLASTN, BLASTX (Altschul et al) ·, J. Mol. Biol. 2 1 5:403 (1 990)) and DNASTAR (DNASTAR, Inc., 1228 South Street, Madison City, Wisconsin, USA). In the context of this case, it should be understood that when using sequence analysis software for analysis, the results of this analysis will be based on the "predetermined number" of the reference program, unless otherwise stated. The “Preset Number” used here will represent any number or parameter group originally built in the -122- 201207108 when the software is first enabled. As far as the sequence of DNA is concerned, "chemical synthesis" means that the nucleotides of the components are composed in a test tube. Manual chemical synthesis of DN A can be accomplished using well-developed methods or automated chemical synthesis using one of a variety of commercial machines. Thus, the gene can be tailored to the optimal gene expression based on the optimized nucleotide sequence to reflect the codon bias of the host cell. The skilled person knows that if the use of the crypto code is biased towards the host's preferred codon, it will likely be successful. The preferred codon determination can be based on a genetic investigation of the source of the host cell I) from which sequence information is available. As used herein, two or more of the respective operable gene regulatory systems are referred to as "orthogonal", and when a) each given system is regulated by an individual ligand of a selected concentration, the gene expression amplitude of the system is measurable. Alteration, and b) the alteration system is significantly different from all other changes in the performance of the operability system in the cell, tissue or organism, whether the actual regulation occurs simultaneously or sequentially. Preferably, modulation of each respective operability gene regulation system y results in a change in gene expression that is at least 2-fold higher than, for example, at least 5 times higher than all other operability systems in a cell, tissue or organism, 10 times, 100 times or 500 times. Ideally, modulating the performance of a given system by an individual ligand of a selected concentration results in a measurable change in the gene expression amplitude of the system' and the performance of all other operability systems in the cell, tissue or organism is not measurable Change. In this case, the multiple inducible gene regulatory system is referred to as "fully orthogonal". Useful orthogonal ligands and orthogonal receptor basal gene expression systems are described in US 2002/0110861 A1. The term "exogenous gene" refers to a gene that is foreign to an individual, that is, a gene, a non-mutated version of an endogenous mutant gene or a mutant version of an endogenous non-mutation introduced into the individual via the -123-201207108 transformation process. gene. The method of transformation is not critical to the invention and can be any method known to those skilled in the art to be suitable for the individual. The exogenous gene may be a natural or synthetic gene introduced into the individual in the form of DNA or RNA, which may act via a DNA intermediate such as produced by reverse transcriptase. The genes can be introduced into the target cells, introduced directly into the individual, or introduced indirectly by transferring the transformed cells into the individual. The term "therapeutic product" refers to a therapeutic polypeptide or therapeutic polynucleotide that confers a beneficial function to the host cell in which the product is expressed. Therapeutic polypeptides can include, but are not limited to, peptides of only three amino acids in length, single or multi-stranded proteins, and fusion proteins. Therapeutic polynucleotides can include, but are not limited to, antisense oligonucleotides, small interfering RNAs, ribozymes, and RNA external leader sequences. The therapeutic product may comprise a naturally occurring sequence, a synthetic sequence or a combination of natural and synthetic sequences. The term "ligand-dependent transcription factor complex" or "LDTFC" refers to a transcription factor comprising one or more protein subunits that modulate gene expression driven by a "factor-regulated promoter" as defined herein. . The LDTFC model "ecdysone receptor complex" generally refers to a heterodimeric protein complex with at least two nuclear receptor family members, namely the ecdysone receptor ("EcR") and the super-valve protein ("USP"). (See Yao et al., Nature 366:476 (1 993)); Yao et al., Cell 7 1:63 (1 99 2)). Functional LDTFCs such as EcR complexes may also contain additional proteins such as immunophilins. Additional members of the protein nuclear receptor family known as transcription factors (such as DHR38, PFTZ-1 or other Kun-124-201207108 paralogs) may also be ligand-dependent or non-matching of EcR and/or USP. Body dependence. LDTFCs such as the EcR complex may also be heterodimers of the EcR protein to the vertebrate homolog of the super-valve protein, the retinoic acid-X-receptor ("RXR") protein or the chimera of USP and RXR. The terms "LDTFC" and "EcR complex" also encompass homodimeric complexes of EcR protein or USP, as well as single polypeptides or trimers, tetramers and other polymers having the same function. The LDTFC, such as the EcR complex, can be activated by an active dermatodermal steroid or a non-sterol ligand that binds to one of the proteins of the complex, the protein comprising EcR but not excluding other proteins of the complex. LDTFCs such as EcR complexes include proteins belonging to members of the nuclear receptor superfamily, in which all members are characterized by the presence of one or more polypeptide subunits, including an amine-terminal transactivation domain ("AD" ", "TD" or "TA"), DNA binding domain ("DBD") and ligand binding domain ("LBD"). The AD may exist in a form of fusion with "heterodimer" or "HP". The fusion protein comprising AD and HP of the present invention is referred to herein as "co-activated protein" or "0-8". DBD and LBD can be expressed in the form of fusion proteins, which are referred to herein as "ligand-inducible transcription factors ("LTF"). The fusion partners can be distinguished by a linker such as a hinge. Some members of the LTF family also have another transactivation domain on the carboxyl end side of LBD. The DBD is characterized by having two zinc cysteine fingers in between which are two amino acid phantom P boxes and D boxes, which confer ecdysone response element specificity. These domains may be chimeras of natural domains, modified domains or different domains of heterologous receptor proteins. -125- 201207108 DNA sequences constituting exogenous genes, response elements and LDTFCs such as EcR complexes may be incorporated into archaebacteria, prokaryotic cells such as Escherichia coli, Bacillus subtilis or other enteric Bacteria, or eukaryotic cells such as plant or animal cells. However, since many proteins expressed by genes are improperly treated in bacteria, eukaryotic cell lines are preferred. These cells may be in the form of single or multicellular organisms. The nucleotide sequences of the exogenous gene, response element and receptor complex may also be incorporated into an RNA molecule, preferably in the form of a functional viral RNA, such as tobacco mosaic virus (tobacco m o s a i c v i r u s ). In the case of eukaryotic cells, vertebrate cell lines are preferred because they naturally lack molecules that confer a ligand to the EcR of the present invention. Thus, they are "substantially insensitive" to the ligands of the invention. Thus, the physiological or other effects of the ligands useful in the present invention on transformed cells or the entire organism will be ignored. Thus, the cells can grow and exhibit the desired product, substantially unaffected by the presence of the ligand itself. The term "ecdysone receptor complex" generally refers to a heterodimeric protein complex with at least two nuclear receptor family members, namely the ecdysone receptor ("EcR") and the super-valve protein ("USP"). (See Yao et al., Nature 366: 476 (1993); Yao et al., Cell 71: 63 (1992)). Functional EcR complexes may also contain additional proteins such as immunophilins. Additional members of the nuclear receptor family of proteins known to be transcription factors (such as DHR38, PFTZ-1 or other insect homologs) may also be ligand-dependent or non-ligand-dependent partners of EcR and/or USP. The EcR complex may also be a heterodimerization of an EcR protein to a vertebrate homolog of a super-valve protein, a retinoic acid-X-receptor ("-126-201207108 RXR") protein, or a chimera of USP and RXR. body. The term EcR complex also comprises an EcR protein or a homodimeric complex of USP.

The EcR complex can be activated by an active ape steroid or a non-sterol ligand that binds to one of the proteins in the complex, the protein comprising EcR but not excluding other proteins of the complex. The term "ligand" as used herein, when applied to an EcR-substrate gene switch, describes small and soluble molecules that activate a gene switch to stimulate the expression of the polymorphism encoded therein. Examples of ligands include, but are not limited to, ecdysteroids, such as ecdysone, 20-hydroxy quercetin (|indolone, ponasterone A, midazorone A, and the like, 9-cis-retinoic acid) , a synthetic analog of retinoic acid, hydrazine, Ν'-dimercaptopurine, such as those in U.S. Patent Nos. 6,013,836, 5,117,057, 5, 530,028 and 5, 378, 726, and U.S. Published Application Nos. 2005/02092 8 3 and 2006/00201 46 U.S. Patent Application Serial No. 2004/0 1 7 1 65 1; oxadiazoline; benzoyl cyanoalkyl cyanohydrazine, such as disclosed in European Application No. 46 1,809; Ν, Ν'-diaryl fluorenyl hydrazine, such as disclosed in U.S. Patent No. 5,225,443; U Ν-mercapto-fluorenyl-alkylcarbonyl hydrazine, such as disclosed in European Application No. 234,994; Ν-aryl fluorenyl- Ν-Alkyl-Ν'-aryl fluorenyl hydrazine, such as those described in U.S. Patent No. 4,98,5, 461; decyl ketones, such as those described in U.S. Patent Application Publication No. 2004/004903, and the like; Including 3,5-di-tertiary butyl-4-hydroxy-indole-isobutyl-benzamide, 8-0-Ethyl hydrazone, oxysterol, 22(R) hydroxycholesterol, 24 (S) hydroxyl Sterol, 25-epoxycholesterol, T090 1 3 1 7 , 5·α-6-α-epoxycholesterol-3-sulfate (ECHS), 7-ketocholesterol-3-sulfate, farnesol , bile acid, 1,1-bisphosphonate, phytohormone III and the like. Examples of dimercaptopurine-127-201207108 which can be used in the present invention include RG-ll 581 9 (3,5-II) Methyl-benzoic acid NU-ethyl-2,2-dimethyl-propyl)-:^'-(2-methyl-3-methoxy-benzhydryl)-indole), RG- 115932((R)-3,5--methyl-benzoic acid N-(l-secondary Γ&quot;yl-butyl)-&gt;^'-(2-ethyl-3-methoxy-benzene醯基)-醯肼) and 11〇-115830(3,5-dimethyl-benzoic acid N-(l-tert-butyl-butyl)-Ν'-(2-ethyl-3-methoxy Base-benzhydryl)-醯肼). Other bismuths that can be used in the practice of the present invention are described in U.S. Patent Application Serial No. 1 2/155,111, filed on May 29, 2008, and PCT/US2008/006757, filed on May 29, 2008.

The EcR complex includes proteins belonging to members of the nuclear receptor superfamily, all of which are characterized by an amine-terminal transactivation domain ("TA"), a DNA binding domain (DBD), and a hinged partition. Body binding domain (LBD). Some family members also have another transactivation domain on the carboxy-terminal side of LBD. The DBD is characterized by having two zinc cysteate fingers with a two amino acid phantom P box and a D box in between, which confers specificity to the cutaneous hormone response element. These domains may be natural domains, modified domains or chimeras of different domains of heterologous receptor proteins. The DNA sequences of exogenous genes, response elements and EcR complexes may be incorporated into archaea ( Archaebacteria), prokaryotic cells such as Escherichia coli, Bacillus subtilis or other enteric bacteria, or eukaryotic cells such as plant or animal cells. However, since many proteins expressed by genes are incorrectly treated in bacteria, eukaryotic cell lines are preferred. These cells may be in the form of single or multicellular organisms. Nucleotides of exogenous genes, response elements and receptor complexes -128-201207108 Sequences may also be incorporated into RNA molecules, preferably in the form of functional viral RNA, such as tobacco mosaic virus . In the case of eukaryotic cells, vertebrate cell lines are preferred because they naturally lack molecules that confer a ligand that reacts to the EcR of the present invention. Therefore, they are "substantially insensitive" to the ligands of the present invention. Thus, the physiological or other effects of the ligands useful in the invention on transformed cells or the entire organism will be ignored. Thus, the cells can grow and express the desired product, substantially unaffected by the presence of the ligand itself. (Λ) When an EcR ligand is used with an EcR complex and in turn binds to a response element linked to an exogenous gene (eg, IL-12), the EcR ligand provides an external temporal regulation of the expression of the exogenous gene. The order of binding of the various components to each other, ie, the order in which the ligand-receptor complex and the receptor complex are combined with the reactive element, is not critical. Typically, the expression of the exogenous gene is regulated by the EcR complex and specific control. Or a binding of a regulatory DNA element. The EcR protein has at least three domains, such as a transactivation domain, a DNA binding domain, and a ligand binding domain, as other members of the nuclear receptor family. A subgroup of the receptor family also has a region that is less well defined to cause heterodimerization properties. After the ligand binding domain of the EcR protein is heterodimerized with the USP or RXR protein, the ligand and the ligand The combination of the binding domains allows the DNA-binding domain of the activated form of the heterodimeric protein to bind to the response element, thereby causing the expression or inhibition of the exogenous gene. Binding to EcR or USP and resulting in the formation of an active homodimeric complex (eg, EcR + EcR or USP + USP). In one embodiment, the one or more receptor domains may be non-129- 201207108 is the same as generating a chimeric gene switch. Typically, the plurality of domains can be selected from sources other than the source of the other domain such that the $@ sex receptor is in the selected host cell or organism. The transactivation of the living axillary 'complementary binding to the ligand and the identification of the specific reaction element is optimal &amp; ° In addition, the reaction element itself can be modified or replaced by other DN A binding protein domain reaction elements , such as the GAL-4 protein from yeast (see Sadowski et al, Nature 335: 563 (1988)) or the LexA protein from the large intestine rod (see Brent et al., Cell 43: 729 (1 98 5 )) To accommodate chimeric EcR complexes. Another benefit of chimeric systems is that they are capable of selecting promoters for driving exogenous genes based on the desired end result. These dual controls are particularly important in the field of gene therapy. Especially when the cell When a toxic protein is produced, the cells whose expression timing and performance occur can be controlled. When the gene is introduced into the cell of the individual in an operably linked manner with an appropriate promoter, the expression of the exogenous gene Is controlled by the presence of a ligand of the invention. The promoter may be constitutively or inducibly regulated' or may be tissue specific (ie, expressed only in a particular cell type) or specific to certain stages of development of the organism. In certain embodiments, the therapeutic switch promoters described in the methods are constitutive. In certain embodiments, the therapeutic switch promoter is under conditions associated with a disease, disorder, or condition. Activating, for example, the promoter may be activated by a disease, by a particular physiological, developmental, differentiated or pathological condition, and/or by one or more specific biological molecules; and/or the promoter is in a particular tissue or cell The type is activated. In certain embodiments, the disease, condition or condition is responsive to a therapeutic polypeptide or polynucleotide. -130-201207108 For example, in certain non-limiting embodiments, the therapeutic polynucleotide or polymorphism can be used to treat, prevent, ameliorate, reduce symptoms, prevent disease progression, or cure the disease, condition, or condition, However, it is not necessary to complete any or all of the above effects. In certain embodiments, the first and second polynucleotides are introduced to allow expression of the ligand-dependent transcription factor complex under conditions associated with the disease, condition or condition. In one embodiment, the therapeutic method is performed such that the therapeutic polypeptide or therapeutic polynucleotide is expressed and disseminated in an individual in an amount sufficient to treat, ameliorate or prevent the disease, condition or condition. As used herein, "dispersion" means that the polypeptide is sufficiently expressed and released from the modified cells to have an effect or activity in the individual. Dissemination can be systematic, local, or any systematic or localized dissemination. For example, the therapeutic polypeptide or therapeutic polynucleic acid can be systemically disseminated via the bloodstream or lymphatic system. Alternatively, the therapeutic polypeptide or therapeutic polynucleotide can be localized in the tissue or organ to be treated. Many genomic Q and c DN A nucleic acid sequences encoding various polypeptides, such as transcription factors and reporter proteins, are well known in the art. Those skilled in the art can obtain nucleic acid sequence information for almost all known genes, and can obtain the nucleic acid molecule directly from a public repository, a facility that publishes the sequence, or can perform routine methods to prepare the molecule. See, for example, the description of the sequence registration number described below. A gene switch may be any gene switch system that regulates gene expression by adding or removing a particular ligand. In one embodiment, the gene switch is a gene switch in which the amount of gene expression is dependent on the amount of ligand present. Examples of ligand-dependent transcription factors useful in the gene switch of the present invention - 131 - 201207108 include, but are not limited to, individual ligands thereof (eg, glucocorticoids, estrogens, luteinizing hormones, retinoids, ecdysones, and Members of the nuclear receptor superfamily activated by their analogs or mimics and rTT A activated by tetracycline. In one aspect of the invention, the gene is associated with an EcR-substrate gene switch. Examples of such systems include, but are not limited to, U.S. Patent No. 6,258,603, U.S. Patent No. 7,045,315, U.S. Patent Application Serial No. 2006/00147 1 1 , 2007/0 1 6 1 086 and International The system described in the publication WO 0 1/708166. Examples of chimeric ecdysone receptor systems are described in U.S. Patent No. 7,09,03,8, U.S. Patent Application Serial Nos. 2002/0110861, 2004/0033600, 2004/0096942, 2005/02664 5 7 and 2006 WO 01/70 8 1 6 &gt 02/29075 and WO 2 005/ 1 0 86 1 7. An example of a non-sterol ecdysone agonist regulatory system is the Rheo Switch® mammalian inducible expression system (New England Biolabs, Ipswich, MA). In one embodiment, the polynucleotide encoding the gene switch comprises a single transcription factor sequence encoding a ligand-dependent transcription factor under the control of a promoter. The transcription factor sequence may encode a ligand-dependent transcription factor of a naturally occurring or artificial transcription factor. An artificial transcription factor is a transcription factor in which the native sequence of the transcription factor has been altered, e.g., by sequence mutation or by combining domains from different transcription factors. In one embodiment, the transcription factor comprises a purine nuclear receptor ligand binding domain (LBD). In a Shi-132-201207108 behavioral pattern, the nucleus receptor LB C line is derived from EcR, ubiquitous receptor, orphan receptor 1, NER-1, steroid hormone nuclear receptor 1, retinoid X receptor. Interaction protein-15, liver X receptor beta, steroid hormone receptor-like protein, liver X receptor, liver X receptor alpha, farnesoid X receptor, receptor interacting protein 14 or farnesol receptor . In another embodiment, the sputum nuclear receptor LBD is derived from a retinoid receptor.

EcR and other sputum groups are members of the nuclear receptor superfamily of the system, and all members are generally characterized by an amine-terminal transactivation domain (TD), a DNA binding domain (DBD), and a DBD with a hinge region. Separate LBD. The term "DNΑ binding domain" as used herein encompasses a DNA binding protein from a minimal polypeptide sequence to a full length DNA binding protein, so long as the function of the DNA binding domain is linked to a particular reaction element. Members of the nuclear receptor superfamily also include four or five domains: A/B, C, D, E, and some members have F domains (see US Patent No. 4,981,784 and Evans, Science 240:8). 8 9 (1 9 8 8)). The "A/B" domain corresponds to the transactivation domain, "C" corresponds to the DNA binding domain, "D" corresponds to the hinge region, and "ΓΕ" corresponds to the ligand binding domain. Some family members also have another transactivation domain on the carboxy-terminal side of the LPD corresponding to "F". The DBD is characterized by having two zinc cysteate fingers with a two amino acid phantom P box and a D box in between, which confer specificity to the reaction element. These domains may be chimeras of natural domains, modified domains or different domains of heterologous receptor proteins. EcR, like a subgroup of the nuclear receptor family, also has a region that is less well defined and that causes heterodimerization. Since the domain of the nuclear receptor is modular in nature, LBD, -133-201207108 DBD and TD may be interchangeable. In another embodiment, the transcription factor comprises a TD, DBD, and a scorpion nuclear receptor LBD that recognizes a response element linked to an exogenous gene to modulate the expression of the exogenous gene. In certain embodiments, the scorpion nuclear receptor LBD comprises a substitution mutation. In another embodiment, the polynucleotide encoding the gene switch comprises a first transcription factor sequence under the control of a first promoter and a second transcription factor sequence under the control of a second promoter, wherein the first transcription is The factor sequence and the protein encoded by the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor, a "dual switch" or "hybrid" substrate gene switch. The first and second promoters may be the same or different. In certain embodiments, the polynucleotide encoding the gene switch comprises a first transcription factor sequence and a second transcription factor sequence under the control of a promoter, wherein the first transcription factor sequence and the second transcription factor sequence are encoded The protein interacts to form a protein complex that functions as a ligand-dependent transcription factor, a "single gene switch." The first transcription factor sequence and the second transcription factor sequence can be joined by an internal ribosome entry site (IRES). The IRES can be g EMCV IRES. In one embodiment, the first transcription factor sequence encodes a polypeptide comprising TD, DBD, and a scorpion nuclear receptor LBD, the DBD identifying a response element linked to the exogenous gene to modulate the expression of the exogenous gene, The second transcription factor sequence encodes a transcription factor comprising a nuclear receptor LBD selected from the group consisting of a vertebrate RXR LBD, an invertebrate RXR LBD, a super-valve protein-134-201207108 LBD or a chimeric LBD comprising two polypeptide fragments. ' wherein the first polypeptide fragment is from a vertebrate RXR LBD, an invertebrate RXR LBD or a super-valve protein LBD, and the second polypeptide fragment is from a different vertebrate RXR LBD, an invertebrate RXR LBD or an ultra-valve Protein LBD. In another embodiment, the gene switch comprises a first transcription factor sequence encoding a first polypeptide and a second transcription factor sequence encoding a second polypeptide comprising a nuclear receptor LBD and a DBD, the DBD Identifying a response element linked to an exogenous gene to modulate the expression of the exogenous gene, the second polypeptide comprising TD and a nuclear receptor LBD, wherein one of the nuclear receptor LBD is a nuclear receptor LBD . In one embodiment, the first polypeptide is substantially free of TD and the second polypeptide is substantially free of DBD. For the purposes of the present invention, &quot;substantially free&quot; means that the problematic protein does not contain sufficient sequence of the problematic domain to provide activation or binding activity. In another aspect of the invention, the first transcription factor sequence encodes a protein comprising a heterodimer with TD, and the second transcription factor sequence encodes a protein comprising QD and LBD. When there is only one nuclear receptor LBD sputum group LBD, the other nuclear receptor LBD may be derived from any other nuclear receptor to form a dimer with the sputum group LBD. For example, when the nucleus receptor LBD is EcR LBD, other nuclear receptor LBD "concomitants" may come from EcR, vertebrate RXR, invertebrate rxr, super-valve protein (USP) or contain at least two different A chimeric nuclear receptor selected from the group consisting of a vertebrate RXR, an invertebrate RXR or a USP nuclear receptor LBD polypeptide fragment (see WO 0 1 /708 1 6 A2, International Patent Application PCT/US02/05235 and US 2004/) 0096942 A1). "Companion" nuclear receptor ligand -135- 201207108 The body binding domain may additionally comprise a truncation mutation, a deletion mutation, a substitution mutation or other modification. In one embodiment, the vertebrate RXR LBD is derived from Homo sapiens, Mus musculus, Rattus norvegicus, Gallus gallus, pig pigs. (Sus scrofa domestica), frog Xenopus laevis, zebrafish zebrafish (Danio rerio), tunicate, Polyandrocarpa misakiensis or Tripedalia cysophora RXR. In one embodiment, the invertebrate RXR ligand binding domain is derived from the super-valve polymorphism ("LmUSP") of Locusta migratoria and the RXR homolog of the Amblyomma americanum. RXR homolog 2 ("AmaRXR2"), RXR homolog (CpRXR), beetle of the genus Crab pugilator RXR homolog of Tenebrio molitor ("TmRXR"), RXR homolog of "Apis mellifera" ("AmRXR"), RXR homolog of Myzus persicae ("MpRXR") or RXR homolog of non-dipterous insect/non-lepidary insects. In one embodiment, the chimeric RXR LBD comprises at least two RXR homologs selected from the group consisting of a vertebrate species RXR polypeptide fragment, a non-vertebrate species RXR polypeptide fragment, or a non-dipterous insect/non-Lepidopteran insect invertebrate species. A polypeptide fragment of a polypeptide fragment. The chimeric RXR ligand binding domain for use in the present invention may comprise at least two different species of RXR polypeptide fragment -136 - 201207108, or when the species are identical, the two or more polypeptide fragments may be derived from the species RXR polypeptide fragment Two or more different isomers. In one embodiment, the chimeric RXR ligand binding domain comprises at least one vertebrate species RXR polypeptide fragment and an invertebrate species RXR polypeptide fragment. In another embodiment, the chimeric RXR ligand binding domain comprises at least one vertebrate species RXR polypeptide fragment and a non-dipteran insect/non-lepidary insect invertebrate species RXR homolog polypeptide fragment. When the ligand binds to the LBD of the nuclear receptor and in turn binds to a response element linked to the exogenous gene, the ligand provides an expression of the external temporal regulation of the exogenous gene. It is not important that the various components of the present invention are combined with each other, i.e., the binding of the ligand to the LBD, the binding of the DBD to the reactive element, the binding of the TD to the promoter, and the like. In a particular embodiment, the ligand binds to the LBD of the sputum nuclear receptor and to the nuclear receptor LBD to allow the exogenous gene to be expressed. This machine does not exclude the possibility that the ligand binds to the sputum nuclear receptor (GHNR) or its partner and results in the formation of an active homodimeric complex (e.g., GHNR + GHNR or concomitant + partner). Preferably, the hybrid gene switch is produced in one or more different receptor domains. Typically, one or more of the three domains DBD, LBD, and TD can be selected from sources other than the source of the other domain such that the hybrid gene and the formed hybrid protein are reversed in the selected host cell or organism. The activation activity, complementary binding to the ligand, and identification of specific reactive elements are optimized. In addition, the reaction element itself can be modified or substituted by other DNΑ binding protein domain reaction elements, such as the GAL-4-137-201207108 protein from yeast (see Sadowski et al., Nature 3 3 5:5 63 ( 1 988)) or LexA protein from E. coli (see Brent et al., Cell 43: 729 (1 98 5)), or specifically for proteins that have been designed, modified, and selected for specific interactions. Synthetic response elements for target interaction (see, eg, Kim et al., Proc. Natl. Acad. Sci. USA, 94:3 6 1 6 (1 997)) to accommodate hybrid receptors. Functional ECR complexes may also contain additional proteins such as immunophilins. Additional members of the nuclear receptor family known as transcription factors (such as DHR38 or PFTZ-1) may also be ligand-dependent or non-ligand-dependent partners of EcR, USP and/or RXR. In addition, other cofactors may be required, such as proteins commonly referred to as coactivators (also known as adaptors or mediators). These proteins do not specifically bind to the DN A sequence and are not involved in basic transcription. They may exhibit their effects on transactivation via various mechanisms, including stimulating DNA binding to activators, affecting chromatin structure, or mediator activator initiation complex interactions. Examples of such auxiliary activators include RIP140, TIF1, RAP46/Bag-1, ARA70, SRC-1/NCoA-1, TIF2/GRIP/NCoA-2, ACTR/AI B 1/RAC 3/p CIP, and hybrid The activator C response element B binds to the protein CBP/p3 00 (see retrospective article Glass et al., Curr. Opin. Cell Biol. 9:222 (1 997)). In addition, protein cofactors, often referred to as co-repressors (also known as repressors, silencers, or silent mediators), may be required to effectively inhibit transcriptional activation in the absence of a ligand. These co-repressors can interact with the ligand-free EcR to silence the activity of the response element. Current evidence suggests that ligand binding alters the conformation of the receptor, resulting in the release of co-repressors and the attraction of the above-mentioned co-activators, thereby eliminating their silent -138-201207108 activity. Examples of co-repressors include N-CoR and SMRT (see review literature Horwitz et al., Mol Endocrinol. 1 0: 1 1 6 7 (1 9 9 6)). These cofactors may be endogenous factors in the cell or organism, or may be exogenously added in a regulated or unregulated manner. The exogenous gene line is operably linked to a promoter comprising at least one response element recognized by the DBD of the ligand-dependent transcription factor encoded by the gene switch. In one embodiment, the promoter comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 sets of reaction elements.

The promoter comprising the desired response element of CI can be a naturally occurring promoter or an artificial promoter produced using techniques well known in the art, e.g., one or more reactive elements are operably linked to a minimal promoter. A gene encoding an immunomodulatory agent such as IL-12, TNF-ct, a signal peptide or any of the transcription factors described herein can also be codon-optimized. In one embodiment, the coding region of an immunomodulatory agent such as IL-12, TNF-[alpha], signal peptide or transcription factor is codon-optimized for human expression. As will be appreciated by those skilled in the art, different nucleic acid coding regions will encode the same polypeptide because of the redundancy of the genetic code. Deviations in the nucleotide sequence of a codon comprising an amino acid encoding any polypeptide chain allow for the variation of the sequence encoding the gene. Since each codon is composed of three nucleotides and the nucleotides that make up DN A are restricted to four specific bases, there are 64 possible combinations of nucleotides, 61 of which encode amino acids ( The remaining 3 codon codes terminate the translation signal). The "gene code" showing which amino acid is encoded by the codon is reproduced in Table 4 herein. As shown in the table, many amino acids are specified by more than one codon. For example, amino acid alanine and proline are encoded by four-139-201207108 triplet codes, and serine and arginine are encoded by six triplets, whereas tryptophan and methionine Then it is encoded by only one triplet. This simplification allows for a wide variety of DNA base compositions without altering the amino acid sequence of the polypeptide encoded by the DNA. Table 4: Standard Gene Cryptography TCAG TTT Phe (F) TCT Ser (S) TAT Tyr (Y) TGT Cys (C) TTC " TCC " TAC " TGC T TTA Leu (L) TTCA " TAATer TGA Ter TTG " TCG " TAG Ter TGG Trp (W) CTT Leu (L) CCT Pro (P) CAT His (H) CGT Arg (R) CTC “ CCC “ CAC “ CGC “ C CTA' CCA11 CAA Gin (Q) CGA “ CTG “ CCG “ CAG "CGG" ATT lie (I) ACT Thr (T) ΑΛΤ Asn (N) AGT Ser (S) ATC " ACC " AAC " AGC " A ΑΤΑ " ACA " AAA Lys ( K ) AGA Arg ( R ) ATG Met ( M) ACG "AAG " AGG " GTT Val (V) GCT Ala (A) GAT Asp (D) GGT Gly (G) GTC " GCC " GAC " GGC " G GTA " GCA " GAA Glu ( E ) GGA " GTG " GCG "GAG "GGG" It is to be understood that any polynucleotide encoding a polypeptide in accordance with the present invention is within the scope of the invention, regardless of the codon used. Many organisms exhibit bias using specific codons to grow The polypeptide chain encodes a specific amino acid. The codon preference or codon bias is the difference in the use of codon between organisms. The simplification of the gene code is widely documented among many organisms. Codon bias is usually related to the translation efficiency of the messenger RN A (mRNA), which is then considered to depend inter alia on the -140-201207108 translated codon. Characteristics and availability of specific transfer RNA (tRNA) molecules. The advantage of intracellular selected tRN A is usually reflected in the most commonly used codons in peptide synthesis. Therefore, genes can be pruned to be based on a given organism. Codon optimization for optimal gene expression. Polynucleotides are prepared by incorporating codons that are preferred in the genes of a given species into the DNA sequence. Due to the large number of species available from a wide variety of animal, plant and microbial species. Gene sequences, so it is possible to calculate the relative frequency of codon usage. Codon usage tables can be easily obtained, for example, on the website http://www.kazusa_or.jp/codon/ (visited on May 30, 2006) The "Cryptographic Use Database", which can be adjusted in a number of ways. See Nakamura, Y., et a 1., &quot;Codon usage tabulated from the international DNA sequence databases: status for the year 2 0 0 0&quot; Nucl. Acids Res. 28:292 (2000). From GenBank

The version of the codon usage table calculated by Release 15 1.0 is copied as the following list 5 (from the same http://www.kazusa.or.jp/codon/). These tables use mRNA nomenclature, so these tables do not use thymine (T) found in DNA, but use uracil (U) found in RNA. These tables are adjusted so that the frequency is calculated for each amino acid, not for all 64 codons. -141 - 201207108 Table 5: Codon usage of human gene (Homo sapiens) Epiformyl acid codon usage frequency phenylalanine (Phe) UUU 0.4525 UUC 0.5475 leucine (Leu) UUA UUG 0.0728 0.1266 CUU 0.1287 CUC 0.1956 CUA 0.0700 CUG 0.4062 Isoleucine (lie) AUU 0.3554 AUC 0.4850 AUA 0.1596 Methionine (Met) AUG 1.0000 Proline (Val) GUU 0.1773 GUC 0.2380 GUA 0.1137 GUG 0.4710 Serine (Ser) UCU 0.1840 UCC 0.2191 UCA 0.1472 UCG 0.0565 AGU 0.1499 AGC 0.2433 Proline (Pro) CCU 0.2834 CCC 0.3281 CCA 0.2736 CCG 0.1149 Threonic acid (Thr) ACU 0.2419 ACC 0.3624 ACA 0.2787 ACG 0.1171 Alanine (Ala) GCU 0.2637 GCC 0.4037 GCA 0.2255 GCG 0.1071 Tyrosine (Tyr) UAU 0.4347 UAC 0.5653 Histamine (His) CAU 0.4113 CAC 0.5887 Glutenin (Gin) CAA 0.2541 CAG 0.7459 Aspartic Acid AAU 0.4614 (Asn) AAC 0.5386 -142- 201207108 Amino Acid codon usage frequency of amino acid _ AAA AAG 0.4212 0.5788 Aspartic acid (Asp) GAU GAC 0.4613 0.5387 glutamic acid (Glu) GAA 0.4161 GAG 0.5839 Cysteine (Cys) UGU 0.4468 UGC 0.5532 Tryptophan (Tro) UGG 1.0000 Arginine (Arg) CGU CGC 0.0830 0.1927 CGA 0.1120 CGG 0.2092 AGA 0.2021 AGG 0.2011 Glycine (Gly) GGU 0.1632 GGC 0.3438 GGA 0.2459 GGG 0.2471

By using these or similar tables, one of ordinary skill in the art can apply these frequencies to any given polypeptide sequence to produce a nucleic acid fragment of a codon-optimized coding region that uses the most for a given species. The codon is optimized to encode the polypeptide. A variety of methods are available for the synthesis of codon-optimized coding regions designed by any of the above methods, including standard and routine molecular biological manipulations well known to those of ordinary skill in the art. In one embodiment, the coding region encoding an immunomodulatory agent, such as TNF-[alpha], in the vector of the invention is codon-optimized. In another embodiment, the coding region is codon optimized for human performance. In a particular embodiment, the TNF-[alpha] of the invention is encoded by a codon-optimized nucleic acid sequence. In order to introduce a polynucleotide into a cell in vivo or in vitro, a vector can be used. The vector may be, for example, a plastid vector or a single or double stranded RN A or -143-201207108 DN A viral vector. By well-known techniques for introducing DNA and RN A into cells, such vectors can be introduced into cells of an individual in need of the vector, such as a mammal. Viral vectors can be either replication or replication defective. In the case of replication-defective viral vectors, viral proliferation will typically only occur in complementary host cells. The term "host cell" or "host" as used herein refers to a cell comprising one or more polynucleotides of the invention in the present invention. Therefore, the vector must at least comprise a polynucleotide of the invention. Other components of the vector may include, but are not limited to, a selectable marker, a chromatin-modifying domain, additional promoters that drive other polypeptides (eg, lethal polypeptides) that may also be present in the vector, genomic embedding sites, recombination sites, and molecular insertions. Pivot. The vector may contain any number of such additional elements' whether or not located within the polynucleotides, such that the vector can be tailored to meet the particular objectives of the method of treatment desired. In an embodiment of the present invention, the vectors introduced into the cells further comprise a "selective marker gene", and when the selectable marker gene is expressed, the gene switch construct of the present invention has been inserted into the genome of the host cell. . Therefore, the selective gene can be used as a positive marker for genome embedding. The selectable marker gene, while not essential to the method of the invention, has its potential to allow the practitioner to select a population of cells in which the vector construct has been inserted into the genome of the cell. Accordingly, certain embodiments of the invention comprise selecting cells that have been successfully inserted into a vector. As used herein, the term "select" or its variants, when used with a cell, is intended to mean a standard, well-known method of selecting cells having a particular genetic composition or phenotype. Typical methods include, but are not limited to, culturing cells in the presence of antibiotics such as G418, neomycin, and ampicillin-144-201207108. Examples of other selectable marker genes include, but are not limited to, genes that confer dihydrofolate reductase, hygromycin or mycophenolic acid resistance. Other selection methods include, but are not limited to, a selectable marker gene that permits the use of thymidine kinase, hypoxanthine-guanine-transphosphokinase or adenine-transphosphokinase as a selector. Cells containing a vector construct of an antibiotic resistance gene will be tolerant to antibiotics in culture. Conversely, cells containing a vector construct of an antibiotic resistance gene will not be able to tolerate antibiotics in culture. As used herein, "chromatin-modifying domain" (CMD) refers to a nucleotide sequence that interacts with various proteins (such as, but not limited to, DNA insulators) that are involved in maintaining and/or altering chromatin structure. See Ciavatta et al., Proc. Nat'l Acad. Sci. U.S.A., 1 0 3:9958 (2006). Examples of CMD include, but are not limited to, chicken beta-globin insulators and chicken high sensitive sites 4 (cHS 4). For example, the use of different CMD sequences between one or more gene programs (ie, promoter, coding sequence, and 3' regulatory region) facilitates the use of differential CMD DNA sequences as "minimum homology arms" with various microorganisms or test tubes. A combination of internal reorganization engineering techniques to "swap" genetic programs between existing multiplex genes and single gene shuttle vectors. Examples of other chromatin modifying domains are known in the art or can be readily identified. The polynucleotide and nucleic acid coding region in the vector of the present invention may be linked to an additional coding region which encodes a secretion or signal peptide to direct secretion of an immunomodulatory agent such as TNFa. According to the signal hypothesis, a protein secreted by a mammalian cell has a signal peptide or a secretory leader sequence, and once the growing protein chain begins to be exported through the rough endoplasmic reticulum, the signal peptide or the leader sequence is excised from the mature protein. Polypeptides secreted by vertebrate cells typically have a signal peptide fused to the N-terminus of the polypeptide which excises from the intact or "full length" polypeptide to produce a secreted or "mature" polypeptide. In one embodiment, the vector of the present invention comprises a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence a polynucleotide encoding a ligand-dependent transcription factor, and (2) a protein encoding one or more proteins having the function of an immunomodulatory agent operably linked to a promoter activated by the ligand-dependent transcription factor, wherein A polynucleotide encoding one or more proteins having the function of an immunomodulatory agent further comprises a nucleic acid sequence encoding a signal peptide. In another embodiment, the signal peptide increases secretion of an immunomodulatory agent, such as TNF-[alpha], encoded by the vector, as compared to a native signal peptide gene comprising an immunomodulatory agent, such as a TNF-[alpha] wild type signal peptide gene. Carrier. In particular, the signal peptides used in the present invention are cryptographically optimized. In a specific embodiment, the signal peptide is encoded by an IL-2 wild type signal peptide gene. In another specific embodiment, the signal peptide is encoded by a codon-optimized IL-2 signal peptide gene. The vector of the present invention may comprise various regulatory regions, such as a 5' non-translated region (5' UTR), 3_ UTR or both. The invention also relates to the use of various regulatory regions to induce enhanced secretion, protein translation, post-translation, mRNA transcription or post-transcriptional processing. The "5' untranslated region" or "5' UTR" of a gene used herein is understood to be a portion of a gene that is transcribed into a primary RNA transcript (pre-mRNA) and which is located upstream of the coding sequence. The primary transcript is the starting RN A product produced by DNA transcription, which contains introns and exons. Xu-146- 201207108 Multiple primary transcripts must be treated with RN A to form physiologically active RN A species. Treatment to become mature mRN A may involve trimming the ends, removing introns, capping, and/or excising individual ribosomal RNA (rRNA) molecules from their RNA precursors. The 5' UTR of the mRNA is therefore the portion of the mRNA that is not translated into a protein and is located upstream of the coding sequence. In the genomic sequence, the 5' UTR is usually defined as the region between the transcription start site and the start codon. The 5' non-translated region (5' UTR) of vertebrate mRNA can range from tens of bases to hundreds of assays (Crowe et al., 2006 BMC Genomics 7:16). A 5'UTR as used herein may be naturally occurring or modified to include one or more nucleic acid sequences that are discontinuous in nature (chimeric sequences), and/or may comprise substitutions, insertions and deletions, and combinations thereof. . In one embodiment, the 5' UTR sequence is derived from a wild type TNF-[alpha] sequence or a 5U2 sequence. In another embodiment, the 5&apos; UTR sequence is a 5&apos; UTR of 5U2. In some embodiments, the 5'UTR induces enhanced protein expression, such as mRNA transcription, pre-transcriptional or post-transcriptional. The 3' non-translated region (UTR) used in the present invention refers to a DNA sequence located downstream (3') of the coding sequence, and may include a polyadenylation [p〇ly(A)] recognition sequence and other coding adjustment signals to affect The sequence of mRN A treatment or gene expression. The polyadenylation signal is typically characterized by affecting the addition of the polyadenylation sequence to the 3&apos; end of the mRNA precursor. Any suitable polyadenylation sequence can be used, including synthetically optimized sequences, as well as BGH (bovine growth hormone), polyoma virus, TK (thymidine kinase), EBV (EB virus), and papilloma virus. Polyadenylation sequences, including human papilloma virus and BPV (bovine papilloma virus). In a specific embodiment, the 3&apos; regulatory region is a polyadenylation sequence of SV40e (human flesh - 147 - 201207108 Tumor virus-40). In another specific embodiment, the &apos;3&apos; regulatory region is a polyadenylation sequence of human growth hormone. In certain embodiments, the signal peptide and/or regulatory region alone or in combination can be increased by at least 2 fold, 3 fold, 4 fold, 5 fold compared to a control without a signal peptide and/or regulatory region. , 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 50-fold, 100-fold, 200-fold, 300-fold, 400-fold or 500-fold protein secretion, transcription or translation. The amount of secretion of a protein such as TNF-α can be normalized to be expressed by a protein encoded by a vector having a wild-type gene. In another specific embodiment of the invention, the single or combined signal peptide and/or regulatory region is increased by from about 5% to about 1%, from about 1% to about 20%, from about 21% to From about 30%, from about 31% to about 40%, from about 41% to about 50%, from about 51% to about 60%, from about 61% to about 70%, from about 71% to About 80%, about 81% to about 90%, about 91% to about 100%, about 101% to about 149%, about 150% to about 199%, The yield of an immunomodulator such as TNF-α is from about 2% to about 29.9%, from about 30,000% to about 499%, or from about 5% to about 10,000%. In a specific embodiment, the invention comprises a vector that conditionally exhibits an immunomodulatory agent, such as TNF-a, wherein the vector comprises a 5UUT-derived 5fUTR, a codon-optimized nucleic acid sequence encoding an IL-2 signal peptide, encoding an immunization A codon-optimized coding region for a modulator such as TNF-a and a polyadenylation signal for SV40e or human growth hormone

In other embodiments, the vector of the invention comprises a strain selected from the group consisting of SEQ ID NO: 47 (vector 43318), SEQ ID NO: 48 (vector 43319), SEQ ID

NO: 49 (vector 43320), SEQ ID NO: 50 (vector 43321), SEQ ID

NO: 51 (vector 43322), SEQ ID NO: 52 (vector 4433), SEQ ID-148-201207108 NO: 53 (vector 43324), SEQ ID NO: 54 (vector 4325), SEQ ID NO: 55 (vector 43326) , a polynucleotide sequence of SEQ ID NO: 56 (vector 43327), SEQ ID NO: 57 (vector 43 3 28) or SEQ ID NO: 58 (vector 43329). In yet another specific embodiment, the vector comprises the polynucleotide sequence of SEQ ID NO: 52 (vector 43 3 23) or SEQ ID NO: 58 (vector 43 329). A particular vector for use in the present invention is a representation vector encoding a protein or polynucleotide. Generally, such vectors comprise a cis-acting control region operably linked to the polynucleotide to be expressed for efficient expression in a host. Suitable trans-acting factors are provided by the host, by a complementary vector or by the vector itself when introduced into a host. A variety of expression vectors can be used to express proteins or polynucleotides. Such vectors include chromosomal, episomal, and viral-derived vectors, for example, derived from bacterial plastids, phage, yeast addenda, yeast chromosomal elements, viruses such as adeno-associated viruses, lentiviruses, baculoviruses, milky vesicles Viruses such as SV40, vaccinia virus, adenovirus, fowlpox virus, pseudorabies virus and retroviral vectors and vectors derived from such combinations, such as those derived from plastid and phage gene elements (eg, viscous bodies and A vector for phage). All can be used in the performance of this aspect of the invention. In general, any vector suitable for maintaining, proliferating or expressing a polynucleotide or protein in a host can be used in this regard. Suitable viral vectors for use in the present invention include, but are not limited to, adenoviral vector vectors, retroviral vectors, herpes simplex virus (HSV) substrate vectors, small virus vector vectors, such as adenovirus-associated virus (AAV) substrate vectors, and -149 - 201207108 AAV-adenovirus chimeric vector. These viral vectors can be prepared using standard recombinant DNA techniques, for example, in Sambrook et al., Molecular Cloning, a Laboratory Manual, 2d edition, Cold Spring Harbor, Press, C o 1 d S pring Harbor, NY _ (1 9 8 9) and the techniques described in Ausube 1 et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley &amp; Sons, New York, NY (1 994). In one embodiment, the viral vector of the invention is an adenoviral vector. Adenovirus (Ad) is a 36 kb double-stranded DNA virus that efficiently transfers DN A to a variety of different target cell types in vivo. Adenoviral vectors can be produced at high cost and can efficiently transfer DN A to both replicating and non-replicating cells. The adenoviral vector genome can be produced using any species, line, subtype, mixture of sp e c i e s, mixtures of multiple lines, mixtures of multiple subtypes or chimeric adenovirus as a source of vector DNA. Adenovirus stocks that can be used as adenoviral sources can be expanded from adenovirus serotypes 1 to 51, which are currently available from the American Type Culture Collection (ATCC, Virginia). Amplification of any other adenovirus serotype provided by any other source from Manassas, Asia. For example, adenoviruses can be subgroup A (eg, serotypes 12, 18, and 31), subgroup B (eg, serotypes 3, 7, 11, 14, 16, 21, 3 4, and 3 5), C. Groups (eg serotypes 1, 2, 5 and 6), subgroup D (eg serotypes 8, 9, 10, 13, 15, 17, 19, 20, 22 to 30, 32, 33, 36 to 39 and 42 To 47), subgroup E (serotype 4), subgroup F (serotypes 40 and 41) or any other adenovirus serotype. Since the human adenovirus serotype-150-201207108 5 (Ad5) genome has been completely sequenced, the adenoviral vector of the present invention described herein pertains to the Ad5 serotype. The adenoviral vector can be any adenoviral vector capable of growing in cells, and a significant portion (although not necessarily substantially) of the vector is derived from or based on the adenovirus-derived genome. The adenoviral vector can be based on the genome of any suitable wild type adenovirus. In certain embodiments, the adenoviral vector is derived from the genome of Group C wild-type adenovirus, particularly serotype 2 or 5. Adenoviral vectors are well known in the art and have been described, for example, in U.S. Patent Nos. 5,559,099, 5,712,136, 5,731,190, 5,837,511 &gt; 5,846,782 &gt; 5,851,806 '5,962,311 '5,965,541, 5,981,225, 5,994,106, 6,020,191 and Patent No. 6,113,913, International Patent Application WO 95/3 467 1, WO 97/2 1 826 and W Ο 0 0 / 0 0 6 2 8 and V ir ο 1 o gy, BN Fields et al., eds. 3d ed., Raven Press, Ltd., New York (1996), Thomas Shenk, Chapter 67 &quot;Adenoviridae and their Replication&quot; and MS Horwitz, Chapter 68 &quot;Adenoviruses&quot;. In other embodiments, the adenoviral vector is a replication defective vector. The term "replication-defective" as used herein means that the adenoviral vector comprises a genome lacking at least one function necessary for replication. A gene defect, a gene function defect, a gene region defect or a genomic region defect system used herein is defined as a function of deleting a sufficient genetic material in a viral genome to destroy or erase a gene whose gene sequence is deleted in whole or in part. The gene function necessary for replication is the function of the genes required for replication (i.e., proliferation) of a replication-deficient adenoviral vector. The necessary gene functions for replication are, for example, early stages of adenovirus (eg, El, E2, and E4 regions), late regions (eg, L1 to L5 regions), genes associated with viral packs -151 - 201207108 (eg, IVa2 gene), and viruses. The associated RNA (eg, VARNA I and/or VA-RNA II) is encoded. In other embodiments, the replication-defective adenoviral vector comprises an adenoviral genome lacking at least one of the replication-necessary gene functions of one or more regions of the adenoviral genome (eg, lacking two or more regions of the adenoviral genome to cause multiple Replication defective adenoviral vector). One or more of the adenoviral genomes are selected from the El, E2 or E4 regions. The replication-deficient adenoviral vector may comprise a defect in the function of at least one replication-required gene of the E1 region (ie, an E1-deficient adenoviral vector), particularly in the regions of the E1A region of the adenovirus and the E1B region of the adenovirus. Defects in gene function. In addition to the defects of this E1 region, the recombinant adenovirus can also have mutations in the main late promoter (MLP), as discussed in International Patent Application WO 00/00628. In certain embodiments, the vector lacks at least one replication essential gene function of the E1 region and at least a portion of the non-essential E3 region (eg, Xba I deletion of the E3 region) (ie, an E1/E3-deficient adenoviral vector) In certain embodiments, the adenoviral vector is "multiple defect", indicating that the adenoviral vector lacks the gene function required for replication of one or more viruses in each of two or more regions of the adenoviral genome. For example, the aforementioned E1-deficient or E1/E 3-deficient adenoviral vector may additionally lack the replication essential gene function of at least one E4 region (i.e., an E1/E4-deficient adenoviral vector). Deletion of the entire E4 region of the adenoviral vector induces a lower host immune response. Alternatively, the adenoviral vector lacks the replication essential gene function of all or part of the E1 region and all or part of the E 2 region (i.e., the E 1 /E2 -deficient gland-152-201207108 viral vector). Adenoviral vectors lacking all or part of the El region, all or part of the E2 region, and all or part of the E3 region's replication essential gene function are also contemplated herein. If the adenoviral vector of the present invention lacks the replication essential gene function of the E2A region, the vector does not comprise a fully deleted E2A region of less than about 23 base pairs in length. Typically, the E2A region of an adenovirus encodes a DBP (DNA binding domain) which is a polypeptide required for DNA replication. Depending on the serotype of the virus, DBP consists of 473 to 529 amino acids. It is generally considered that DBP is an asymmetric protein in the shape of a long ellipsoid which consists of a spherical Ct and an extended Nt domain. Studies have shown that this Ct domain is responsible for the ability of DBP to bind to nucleic acids, bind to zinc, and act on the elongation phase of the DN A chain in DN A synthesis. However, the Nt domain is thought to act on late gene expression in both transcriptional and post-transcriptional phases, responsible for efficient nuclear localization of the protein, and may also be involved in enhancing its own performance. Deletion of the Nt domain between amino acids 2 to 38 indicates that this region is an important part of DBP function (Brough et al., Virology, 1 96, 269-28 1 (1 993)). Although deletion of the E2A region encoding the Ct region of DBP has no effect on viral replication, deletion of the E2A region of amino acids 2 to 38 encoding the Nt domain of DBP prevents viral replication. In one embodiment, the multiplex replication-deficient adenoviral vector comprises an E2A region of the portion of the adenoviral genome. Specifically, for example, the portion of the E2A region to be retained is the E2A region portion of the adenoviral genome defined by the 5' end of the E2A region, particularly the position Ad5 of the E2A region of the adenovirus serotype Ad5 genome ( 23816) to Ad5 (24032). The adenoviral vector may be a replication-deficient gene function lacking only the early region of the adenoviral genome, and a replication-required group of only the late region of the adenoviral genome-153-201207108 due to function and replication of the early and late stages of the adenoviral genome Gene function. The adenoviral vector can also substantially remove the entire adenoviral genome, in which case at least the complete viral inverted terminal repeat (ITR) and one or more promoter or viral ITR and packaging signals (ie adenovirus proliferators) are maintained. . The larger the removed genomic region of the adenovirus, the larger the fragment of the exogenous nucleic acid sequence that can be inserted into the genome. For example, if the adenoviral genome is 36 kb, the intact viral ITR and one or more promoters are retained, and the exogenous insertion ability of the adenovirus is about 35 kb. Alternatively, a multi-defective adenoviral vector comprising only ITR and packaging signals effectively allows insertion of a source nucleic acid sequence of about 37 to 38 kb. Of course, the inclusion of a spacer element in any or all of the defective adenoviral regions will reduce the ability of the adenoviral vector to accommodate large insertion sequences. Suitable replication-defective adenoviral vectors, including multiplex-defective adenoviral vectors, are disclosed in U.S. Patent Nos. 5,851,8,06, 5,99,106, and International Patent Application No. WO 95/3467, and WO 97/21,826. In one embodiment, the carrier used in the method of the invention is described in International Patent Application No. PCT/US0 1/20536. It will be appreciated that deletion of different regions of the adenoviral vector can alter the immune response of the mammal. In particular, deletion of different regions reduces the inflammatory response of adenoviral vectors. Alternatively, the adenoviral vector coat protein can be modified to reduce the ability of the adenoviral vector to be or cannot be recognized by neutralizing antibodies against wild-type coat proteins, as described in International Patent Application WO 98/405. When the adenoviral vector has multiple replication defects, particularly the replication essential gene function of the E1 and E4 regions, the vector may include a spacer element to provide -154-201207108 virus growth in a complementary cell line, similar to a single replication-deficient gland The viral vector is in particular a virus growth achieved by an adenoviral vector containing a defect in the E1 region. The spacer element can comprise a sequence of any desired length. The spacer element sequence may be a coding or non-coding of the adenoviral genome and a natural or non-native sequence, but does not restore the replication necessity function of the defective region. In the absence of a spacer, the fibrin production and/or viral growth of the multiplex replication-deficient adenoviral vector is reduced compared to the single replication-deficient adenoviral vector. However, inclusion of a spacer in at least one region of the defective adenoviral region, preferably in the E4 region, compensates for this fibrin production and reduction in viral growth. The use of spacers in adenoviral vectors is described in U.S. Patent No. 5,851,806. The construction of adenoviral vectors is well known in the art. The adenoviral vector can be constructed and/or purified by the methods described above, for example, as described in U.S. Patent No. 5,965,350, the disclosure of which patents, . The production of adenoviral gene transfer vectors is well known in the art and involves the use of standard molecular biology techniques such as, for example, Sambrook et al. (ibid.), Watson et al. (ibid.), Ausubel et al. (ibid.) And as described in several other documents mentioned herein. A replication-deficient adenoviral vector is typically produced in a complementary cell line that provides an appropriate amount of gene function that is not present in a replication-deficient adenoviral vector but is required for viral proliferation to produce a high-virgin viral vector. liquid. In one embodiment, the cell line complements at least one and/or all of the replication essential gene functions that are not present in the replication defective adenovirus. The complementary cell line may complement at least one of the defects of the replication essential gene-155-201207108 encoded by the early region, the late region, the viral packaging region, the virus-associated RNA region, or a combination thereof, including all adenoviral functions ( For example, an adenovirus proliferator comprising minimal adenoviral sequences, such as only inverted terminal repeats (ITR) and packaging signals or only ITR and adenoviral promoters, is propagated). In another embodiment, the complementary cell line complements at least one of the E1 regions of the adenoviral genome with defects in the function of a replication essential gene (eg, two or more replication essential gene functions), particularly in each El A and E1 B region The defect of copying the essential gene function. Furthermore, the complementary cell line may complement the defect of at least one of the functions of the E6 (especially adenovirus DN A polymerase and terminal protein) and/or E4 region of the adenoviral genome. Deliberately, the defective E4 region-deficient cells contain the E4-ORF6 gene sequence and produce the E4-ORF6 protein. The cell deliberately contains at least ORF6 but no other ORF of the E4 region of the adenoviral genome. A preferred other feature of the cell line is the inclusion of a complementary gene that is not repeated with the adenoviral vector, thus minimizing and virtually eliminating the possibility of vector genome recombination with cellular DN A. Therefore, the probability of the presence of a replicating adenovirus (RCA) can be minimized, even if it cannot be avoided in the carrier preservation solution, so that the carrier preservation solution is suitable for certain therapeutic purposes, particularly gene therapy. The RC A-free vector preservation solution prevents the adenoviral vector from replicating in non-complementary cells. The construction of complementary cell lines involves standard molecular biology and cell culture techniques, such as those described by Sambrook et al. (supra) and Ausubel et al. (supra). Complementary cell lines for the production of gene transfer vectors (e.g., adenoviral vectors) include, but are not limited to, 2 93 cells (e.g., Graham et al., J. Gen. Virol, 3 6, 59-72 (1 977). ), PER.C6 cells (for example, International Patent Application No. WO 97/003 26 and U.S. Patent Nos. 5,9,9,1, 2, and 6,0 3 3,9 0) and 2 9 3 - 0 RF 6 cells (for example, -156 - 201207108 International Patent Application WO 95/3467 1 and Brough et al., J. Virol, 7, 1, 9206-92 13 (1 997)). Insertion of the nucleic acid sequence into the adenoviral genome (e.g., the E1 region of the adenoviral genome) can be accomplished by known methods, for example, by introducing a unique restriction site at a given position in the adenoviral genome. Retroviral vectors are RNA viruses that are capable of infecting a variety of host cells. When infected, the retroviral genome is embedded in the genome of the host cell and replicates with the DNA of the host cell, thereby sustaining the production of viral RNA and any nucleic acid sequences incorporated into the retroviral genome. In this way, long-term performance of therapeutic factors can be achieved when using fl retrovirus. Retroviruses considered for gene therapy are relatively non-pathogenic, although pathogenic retroviruses are present. When a pathogenic retrovirus such as human immunodeficiency virus (HIV) or human T-cell virus (HTLV) is employed, the viral genome must be carefully altered to eliminate toxicity to the host. The retroviral vector can be altered to make the virus a replication defect. Therefore, retroviral vectors are considered to be particularly suitable for stable gene transfer in vivo. Lentiviral vectors such as HI V-q basal vectors are exemplary retroviruses for gene delivery. Unlike other retroviruses, HIV-based vectors are known to incorporate their passenger genes into non-dividing cells and can therefore be used to treat persistent diseases. The H SV-basal viral vector is suitable for use as a gene transfer vector to introduce nucleic acids into various cell types. The mature H SV virion is constructed as a icosahedral nucleocapsid with a membrane and a viral genome consisting of a 152 kb linear double stranded DNA molecule. Most replication-defective HSV vectors contain deletions to remove one or more immediate early genes to avoid replication. The advantage of the HSV vector is its ability to enter a latent stage of long-term DNA expression, and its -157-201207108 can accommodate a large viral DNA genome of 25 kb of exogenous DNA insertion. Of course, the ability of HSVs to initiate long-term production of exogenous proteins may be a disadvantage in terms of short-term treatment plans. However, the average skilled person in the field has the necessary knowledge to determine the appropriate carrier for a particular situation. HSV-substrate vectors are described, for example, in U.S. Patent Nos. 5,83,7, 5, 2, 5, 84, 7, 8 2, 5, 849, 572 and 5, 804, 4 1 3 and International Patent Application WO 9 1 /0278 8, WO 96 /04394, WO 98/15637 and WO 99/06583. The A A V vector is a viral vector that is of particular interest for use in gene therapy planning. AAV is a DN A virus that has not been found to cause human disease. The AAV genome consists of two genes, rep and cap, which are flanked by inverted terminal repeats (ITRs) containing identification signals for DNA replication and viral packaging. AAV requires co-infection with a helper virus (i.e., adenovirus or herpes simplex virus) or the expression of a helper gene for efficient replication. AAV can be propagated in a variety of host cells, including human cells, anthropoid cells, and rodent cells, depending on the helper virus used. The AAV vector used to administer the nucleic acid sequence is usually deleted by about 96% of the parental genome, leaving only the ITR retained. This eliminates the immune or toxic side effects caused by the presence of viral genes. If desired, the AAV rep protein can be co-administered with the AAV vector to enable the AAV vector to be inserted into the genome of the host cell. Host cells comprising the embedded AAV genome have no change in cell growth or cell morphology (see, e.g., U.S. Patent No. 4,797,396). Therefore, long-term therapeutic factors derived from the Eight Diagrams vector can be used to treat persistent and chronic diseases. The polynucleotide sequences in the expression vector are operably linked to appropriate expression control sequences, including, for example, promoters that direct mRNA transcription. Others -158- 201207108 Representatives of promoters include, but are not limited to, constitutive promoters and tissue-specific or inducible promoters. Examples of constitutive eukaryotic promoters include, but are not limited to, the mouse metallothionein I gene (Hamer et al., J. Mol. Appl. Gen. 1: 273 (1 982)), and the TK promoter of herpesvirus ( McKnight, Cell 31: 355 (1982)), the SV40 early promoter (Benoist et al., Nature 290: 304 (1 98 1)) and the vaccinia virus promoter. Other examples of promoters that can be used to drive expression of a protein or polynucleotide include, but are not limited to, tissue-specific promoters and endogenous promoters of other specific proteins, such as albumin promoters (hepatocytes), pro-insulin promoters ( Pancreatic beta cells) and the analogs. In general, expression constructs will contain priming and termination sites for transcription, as well as ribosome binding sites for translation in the transcribed region. The coding portion of the mature transcript represented by the construct may include a translation initiation codon AUG at the beginning of the polypeptide to be translated and a stop codon (UAA, UGA or UAG) at the tail end. In addition, the construct may include a control zone that regulates and causes performance. In general, these regions will be manipulated by controlling transcription, including, for example, suppressor binding sites and enhancers. Examples of eukaryotic vectors include, but are not limited to, pW-LNEO, pS V2CAT, pOG44, pXTl, and pSG from Stratagene; pSVK3, pBPV, pMSG, and pSVL from Amersham Pharmacia Biotech; Clontech's p C Μ VD s Re d 2 - e xp ress, p I RE S 2 - D s R ed 2, p D s R ed 2 - M ito and pCMV-EGFP. Many other vectors are well known and commercially available. -159- 201207108 A particularly useful carrier comprises a molecular insertion fulcrum for rapid insertion and removal of elements of a genetic program, such as those disclosed in U.S. Patent Application Serial No. 2004/0185556, U.S. Patent Application Serial No. 11/233,246, and International Publications Application WO 2005/040336 and WO 2005/116231. An example of such a carrier is the system described in WO 20 07/03 8276 (Intrexon, Blacksburg, Va.). As used herein, the "gene plan" is a combination of gene elements, including a promoter (P), a presentation sequence (E), and a 3' regulatory sequence (3). Therefore, "PE3" is a genetic program. The components in this genetic program can be easily exchanged between the molecular fulcrums flanking the various components of the genetic program. The molecular fulcrum used herein is defined as a polynucleotide comprising at least two non-variable rare or rare restriction sites arranged in a linear manner. In one embodiment, the molecular fulcrum comprises at least three non-variable rare or rare restriction sites arranged in a linear fashion. Usually any molecular fulcrum does not include rare or rare restriction sites at any other molecular fulcrum within the same genetic program. A homologous sequence of more than 6 nucleotides to which a given restriction enzyme acts is referred to as a "rare J restriction site. However, some 6 bp restriction sites occur less frequently than expected, and these sites are The nucleases within which they are cleaved are referred to as "rare" restriction sites. Examples of rare or rare restriction enzymes include, but are not limited to, AsiS I, Pac I, Sbf I, Fse I, Asc I, Mlu I, SnaB I, Not I, Sal I ' Swa I, Rsr Π, BSiW I, Sfo I, Sgr AI, Afllll, Pvu I, Ngo MIV, Ase I, Flp I, Pme I, Sda I, Sgf I , Srf I, Nru I &gt; Acl I, Cla I, Csp45 I, Age I, Bstl 107 I, B St BI, H pa I ' A at II, EcoR V, Nhe I, Spe I, Avi -160- 201207108 11, A vr 11, M fe I, A fe I, F sp I, ρ ρ η I, S ca I, B sp EI , Nde I, Bfr I, Xho I, Pml I, ApaL I, Kas I, Xma I, BsrB I, Nsi I, Sac II, Sac I, B1 p I, PspoM I, Pci I, Stu I, Sph I, BamH I, Bsu36 I, Xba I, BbvC I, Bgl II, Nco I, Hind III, EcoR I, B srG I and S se8 7 8 1 1. The vector may also comprise a restriction site for a second type of restriction enzyme called homing endonuclease (HE). HE enzymes have large, asymmetric restriction sites (12 to 40 base pairs), and their restriction sites are not common in nature. For example, HE called I-Scel has a restriction site of 18 bp (5'TAGGGATAACAGGGTAAT3, (SEQ ID NO: 28)), which is expected to occur only once in a random sequence of 7xl01() base pairs. This frequency of occurrence is equivalent to only one site in the genome that is 20 times the size of the mammalian genome. The rarity of the HE locus greatly increases the likelihood that the genetic engineer will cut the genetic program without destroying the integrity of the genetic program if the HE site is included in the appropriate location of the vector of the selection vector. Selection of appropriate vectors and promoters for expression in a host cell is a well-known procedure, and the necessary techniques for constructing and introducing the vector into the host, as well as the performance in the host, are routine techniques in the field. The introduction of the polynucleotide into the cell can be a transient transfection, a stable transfection, or can be a locus-specific insertion vector. Transitional and stable transfection vectors into host cells can be achieved by calcium phosphate transfection, DEAE-dextran vector transfection, cationic lipid vector transfection, electroporation, transduction, infection or other methods. These methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods in Molecular B i o 1 o g y (1 9 8 6) &gt; -161 - 201207108

Keown et al., 1 990, Methods Enzymo 1. 1 8 5: 5 27-3 7.

Sambrook et al., 200 1, Molecular Cloning, A Laboratory Manual, Third Edition, Cold Spring Harbor Laboratory Press, N.Y. These stable transfection methods result in random insertion of the vector into the genome of the cell. In addition, the number and orientation of the vectors are generally random. In one embodiment of the invention, the vector is inserted into a biological neutral site of the genome. A biologically neutral site is a site that inserts a polynucleotide into the genome with little or no interference, if any, of the normal function of the cell. Biological neutral sites can be analyzed using available bioinformatics. Many biologically neutral sites are known in the art, such as the R0SA-equal locus. Other biological neutral sites can be identified using routine techniques well known in the art. Characterization of the genomic insertion site is performed using methods known in the art. When introducing a vector into a cell, a method of locus-specific insertion can be used in order to control the position, number of copies, and/or orientation of the polynucleotide. Methods for specific insertion of loci are well known in the art and include, but are not limited to, homologous recombination and recombinant enzyme vector genomic insertion. Of course, if a locus-specific insertion method is to be used in the methods of the invention, such vectors may comprise elements that facilitate the specific insertion of the locus, such as, but not limited to, homologous recombination. For example, such vectors may contain one, two, three, four or more genomic integration sites (GIS). The "genomic integration site" as used herein is defined as the portion of the nucleotide sequence of the vector sequence that is identical or nearly identical to the genomic portion of the cell'. This portion allows the vector to be inserted into the genome. In particular, the vector may comprise two bases - 162 - 201207108 due to at least one of the polynucleotides. Of course, the GIS can be on the side of all other components or even all of the components on the carrier. In another embodiment, locus-specific insertion can be performed by recombinase site-specific gene insertion. Briefly, bacterial recombinases such as, but not limited to, PhiC31 integrase can act on "pseudo" recombination sites within the human genome. These pseudorecombination sites can be targets for locus-specific insertion using recombinases. Recombinase site-specific gene insertion lines are described in Thyagarajan et al., Mol. Cell Biol. 21:3926 (2001). Recombinases that can be used for C) recombinase site-specific gene insertion and other examples of such individual sites include, but are not limited to, serine recombinases such as R4 and TP 9 0 1 -1 and WO 2006/08 3 25 3 described recombinase. In another embodiment, the vector may comprise a chemically resistant gene, such as a multidrug resistance gene mdrl, dihydrofolate reductase or a 〇6-alkylguanine-DN A transaminase. Chemically resistant genes can be under the control of a constitutive (eg, CM V) or inducible (eg, Rheo Switch®) promoter. Q In this embodiment, if it is desired to treat an individual's disease while maintaining the modified cells in the individual, the clinician may use a chemotherapeutic agent to kill the diseased cells, but the modified cells will not be affected by the The effect of the agent is due to the performance of the appropriate chemical resistance gene and can be used continuously to treat, ameliorate or prevent the disease or condition. By placing the chemical resistance gene under the control of an inducible promoter, non-essential chemistry can be avoided. Resistance gene expression, but will continue to be available if the gene is to be treated continuously. If the modified cells themselves become diseased cells, they can still be destroyed by inducing the expression of a lethal polypeptide as described below. -163- 201207108 The method of the present invention can be achieved by introducing a polynucleotide encoding a gene switch and an exogenous gene into a cell of an individual. Any of the known methods for introducing polynucleotides into cells known in the art, such as those described above, can be used. When the polynucleotide is to be introduced into a cell in vitro, the cells can be taken from the individual by any technique known in the art including, but not limited to, biopsy, scraping, and surgical tissue removal. The isolated cells can be cultured for a period of time sufficient to allow introduction of the polynucleotide into the cells, e.g., 2, 4, 6, 8, 10, 12, 18, 24, 36, 48 hours or longer. Methods for culturing primary cells in a short period of time are well known in the art. For example, the cells can be incubated or cultured in a culture dish (e.g., a microplate). In the case of an in vitro therapeutic method, the cell line is isolated from the individual and cultured under conditions suitable for introduction of the polynucleotide into the cell. Once the polynucleotides are introduced into the cells, the cells are cultured for a sufficient period of time to allow for the expression of ligand-dependent transcription factors, such as 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 18 or 24 hours or more. At some point after the polynucleotide is introduced into the cell (whether before or after a significant amount of the ligand-dependent transcription factor is expressed), the cells are introduced back into the individual. The lead can be performed by any method known in the art, such as intravenous infusion or direct injection into tissue or lumen. In one embodiment, prior to the introduction of the cells back into the individual, the presence of polynuclear acid in the cells is determined. In another embodiment, the cell line comprising the polynucleotide is screened (e. g., depending on the presence or absence of a selectable marker for the polynucleotide), and only those cells comprising the polynucleotide are introduced back into the individual. After the cells are introduced back into the individual, the ligand is administered to the individual from -164 to 201207108 to induce expression of the therapeutic polypeptide or therapeutic polynucleotide. In an alternative embodiment, the ligand can be added to the cell even before it is introduced back into the individual such that the therapeutic polypeptide or therapeutic polynucleotide begins to behave before the cell is introduced back. The ligand can be administered by any suitable method, whether systemic (eg, oral, intravenous) or topical (eg, intraperitoneal, intrathecal, intracerebroventricular, direct injection into the cell). Within the tissue or organ). The timing of the desired ligand administration for each cell type and disease or condition can be determined using routine techniques. The in vivo therapeutic method of the present invention involves direct introduction of a polynucleotide, such as an adenoviral vector, into a cell of an individual in vivo. Polynucleotides can be introduced into an individual via systemic or localized (e.g., the site of a disease or condition). Once the polynucleotide is introduced into an individual, the ligand can be administered to elicit the performance of the therapeutic polypeptide or therapeutic polynucleotide. The ligand can be administered by any suitable method, whether systemic (eg, oral, intravenous) or topical (eg, intraperitoneal, intrathecal, intracerebroventricular, direct injection to the onset of 0 disease or Within the tissue or organ of the disease). The ideal ligand administration time for each cell type and disease or condition can be determined using routine techniques. For in vivo use, the ligands described herein can be included in pharmaceutically acceptable carriers such as solutions, suspensions, lozenges, capsules, ointments, elixirs and injectable compositions. The pharmaceutical composition may comprise from 0.01% to 99% by weight of a ligand. The composition can be in single or multiple dose forms. The amount of ligand in any particular pharmaceutical composition will depend on the effective dose, i.e., the dosage required to induce or inhibit the desired gene expression. Suitable routes for administration of the pharmaceutical preparation include oral, rectal, topical-165-201207108 (including transdermal, buccal and sublingual), transvaginal, parenteral (including subcutaneous, intramuscular, intravenous, intratumoral) , intradermal, spinal intrathecal and epidural) 'vitreal and nasal tube. Those skilled in the art will appreciate that the route of administration will depend on the condition being treated and may vary depending on factors such as the condition of the recipient. As used herein, the term "80.111^〇11^2" refers to an adenoviral polynucleotide vector comprising the IL-12 gene under the control of a genetic switch of the RheoSwitch® therapeutic system (RTS), which is capable of activation. Production of IL-12 protein in the presence of a ligand. The term "rAd.cILl2" as used herein refers to an adenoviral polynucleotide control vector comprising the IL-1 2 gene under the control of a synthetic promoter. The term "IL-12p70" as used herein refers to an IL-12 protein which naturally has two subunits commonly referred to as p40 and p35. The term IL-12p70 comprises a fusion protein consisting of two subunits of IL-12 (p40 and p35), wherein the fusion protein may comprise a linker amino acid between the secondary units. [00475] As used herein, the phrase "protein having the function of an immunomodulatory agent" means having at least 20% (eg, at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%). Any biologically active protein of an immunomodulator, the immunomodulator is selected from the group consisting of 11-1, 11^2, 1[-3, 11^4, 11^5, IL-7, IL-8, IL-9, IL -10R or subunits DN, IL-15, IL-18, IL-21, IL-23, IL-24, IL-27, GM-CSF, IFN-α, IFN-γ, CCL3 (MIP-la ), CCL 5 ( RANTES ), CCL7 (MCP3), XCL1 (lymphocyte chemotactic factor), CXCL1 (MGSA-a), CCR7, CCL19 (MIP-3b), CXCL9 (MIG), CXCL 1 0 (IP-1) 0), -166- 201207108 CXCL 1 2 (SDF-1), CCL2 1 (6Ckine), OX40L, 4-1BBL, CD40, CD70, GITRL, LIGHT, b-defensin, HMGB1, Flt3L, IFN-β, TNF -α, dnFADD, BCG, TGF-α, PD-L1, TGFbRII DN, ICOS-L or S100. Similarly, the phrase "protein having the function of IL-12" means having at least 20% (eg, at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%). Any biologically active protein of human IL-1. The biological activities of such immunomodulators are well known. See the table below.

-167- 201207108 Table β immunomodulatory agents I and their functions Immunomodulators Function Interleukin-1 (IL-1) IL-1 is an interleukin produced by activated giant boat cells. IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B cell maturation and proliferation, and fibroblast growth factor activity. IL-1 protein is associated with inflammatory response. Interleukin-2 (IL-2) is a family of interleukins that are produced by T cells that are stimulated by antigenic or cleavage. And other key activities required to regulate the immune response. IL-2 stimulates B cells, mononuclear cells, killer cells activated by lymphocyte mediators, natural killer cells, and glioma cells. Interleukin (IL-3) IL-3 stimulates proliferation of hematopoietic progenitor cells. It is secreted by activated T cells to support the growth and differentiation of bone marrow-derived T cells in an immune response. Studies have shown that the combination of Ad-mIL-3 gene therapy in combination with radiation therapy significantly inhibits tumor growth (Oh 2004). Interleukin-4 (IL-4) IL-4 is an interleukin involved in the activation of at least several B cells and other cell types. IL-4 is a costimulatory factor for DNA synthesis. IL-4 induces the expression of type II MHC molecules on resting B cells. It enhances the secretion of IgE and IgGl and cell surface expression. It also modulates the expression of low affinity Fc receptor (CD23) of IgE on lymphocytes and mononuclear spheres. Interleukin-5 (IL-5) IL-5 stimulates B cell growth and increases immunoglobulin secretion and induces tumor suppression (Nakashima 1993, Wu 1992). Interleukin-7 (IL-7) IL-7 is an interleukin that is a hematopoietic growth factor that stimulates the proliferation of lymphoid progenitor cells. It is important for proliferation in certain stages of B cell maturation. Interleukin-9 (IL-9) IL-9 supports IL-2 independent and IL-4 independent growth of T helper cells. Interleukin-15 (IL-15) IL-15 is an interleukin that stimulates T lymphocyte proliferation. Stimulation of 11^15 requires interaction of IL-15 with IL-2R components, including IL_2R beta and possibly Ιί-2ΙΙγ, but does not require IL-2Ra. -168- 201207108 Immunomodulator Function Interleukin-18 (IL-18) IL-18 enhances the activity of natural killer cells in spleen cells and stimulates the production of interferon-gamma in T-assisted type I cells. Interleukin-21 (IL-21) IL-21 is an interleukin with immunomodulatory activity. IL-21 promotes metastasis between innate and acquired immunity. Interleukin-23 (IL-23) IL-23 acts directly on DC to promote the immune expression of tumor peptides, and can lead to strong intratumoral CD8(+) and CD4(+) T cell infiltration and induce regional lymph nodes and spleen Specific TH1 type response to tumors (Hu 2006). Interleukin-27 (IL-27) IL-27 is a pro-inflamed and anti-inflammatory interferon that regulates T helper cell development, inhibits T cell proliferation, stimulates cytotoxic T cell activity, Inducing homotypic switching in B cells and having multiple effects on innate immune cells. Interleukin-24 (IL-24) IL-24 has been shown to inhibit tumor growth (Susan 2004, Fisher 2003). INF-alpha (IFNa) ffN-alpha has anti-tumor function (Taqliaferri 2005). Interferon beta ΐ (丽Bl) IFNB1 is a member of the interferon protein group that binds to specific cell surface receptors (IFNAR), which stimulate macrophages and natural killer (NK) cells to induce antiviral, antibacterial and anticancer activities. . Interferon gamma, - gamma) IFN-gamma is produced by lymphocytes activated by specific antigens or cleavage substances. In addition to its antiviral activity, IFN-γ also has important immunomodulatory functions. It is a potent activator of macrophages, which has an anti-proliferative effect on transformed cells and enhances the antiviral and antitumor effects of type I interferons. Tumor necrosis factor (TNF-a) TNF-α is mainly secreted by macrophages and induces cell death in certain tumor cell lines. It is a strong pyrogen that causes fever by direct action or by stimulating the secretion of interleukin-1. Chemokine hormone chemotactic hormone (C motif) ligand 1 (XCL1) Chemokine (C motif) ligand 1 (XCL1, also known as lymphocyte chemotactic factor) chemotaxis of CD4+ and CD8+ T cells but not chemotaxis Single-nuclear ball, and induces an increase in intracellular calcium in peripheral blood lymphocytes. The combination of XCL1 with IL-2 and IL-12 enhances immunotherapy and enhances anti-tumor response (Emtage 1999, Wang 2002). CC chemokine ligand 3 (CCL3) CC chemokine ligand 3 (CCL3), also known as macrophage inflammatory protein-l (MIP-l), is the so-called acute inflammatory state of assembly and activation of polymorphonuclear leukocytes Related mononuclear factors (an interleukin produced primarily by mononuclear spheres and macrophages). CCL5 (RANTES) CCL5 (RANTES) is a chemokine of blood mononuclear cells, memory T helper cells, and eosinophils. It causes the basophilic ball to release histamine and activate eosinophils. Combined with CCIU, CCR3, CCR4 and CCR5. It is one of the major HIV inhibitors produced by CD8+ T cells. CC Chemokine Ligand 7 (CCL7) CCL7 is a chemokine that attracts mononuclear and eosinophils but does not attract neutrophils. CCL7 also increases the antitumor activity of mononuclear spheres. The release of gelatinase B was also induced. -169- 201207108 Immunomodulator Function Chemotactic Hormone (CXC Motif) Ligand 9 (CXCL9) CXCL9 is an interleukin that affects the growth, migration or activation of cells involved in immune and inflammatory responses. Chemotactic chemotactic hormone (CXC motif) ligand 10 (CXCL10) Chemokine (CXC motif) ligand 10 (CXCL10) is a small interleukin with chemotaxis Immune system cells, adhesion of T cells to endothelial cells, anti-tumor activity and angiogenesis. Magneto-hormone (cxc motif) Ligand 12 (CXCL12) Chemokine (CXC motif) ligand 12 (CXCL12), also known as stromal cell-derived factor 1 (SDF-1), is a small cell belonging to the myeloid family. Interleukin, a member of the murine family, activates white blood cells and is usually induced by proinflammatory stimuli such as LPS, TNF or IL1. Chemotherapy hormone (C-C motif) receptor 7 (CCR7) CCR7 is a receptor for MIP-3-β chemokine. It is a possible vehicle for the effect of EBV on B lymphocytes or normal lymphocyte function. Chemokine (C-C motif) ligand 19 (CCL19, also known as ΜΙΡ-3β) The role of CCL19 not only in inflammation and immune response, but also in the circulation and homing of normal lymphocytes. CCL19 has important functions for transporting T cells in the thymus and for moving T cells and B cells to secondary lymphoid organs. It specifically binds to the chemokine receptor CCR7. CC Chemokine Ligand 21 (CCL21) CCL21 inhibits hematopoiesis and stimulates chemotaxis. CCL21 is chemotactic for thymocytes and activated T cells in vitro, but is not chemotactic for B cells, macrophages or neutrophils. Interleukin-8 (IL-8) IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T cells but does not attract mononuclear cells. It also involves neutrophil activation. It is released from a variety of cell types in response to inflammatory irritation. Growth factor granules/macrophage colony-stimulating factor (GM-CSF) GM-CSF is an interleukin that stimulates the growth and differentiation of hematopoietic precursor cells of various cell lines, including granules, macrophages, eosinophils, and red blood cells. . FMS-associated tyrosine kinase ligand (FLT3/FLK2 ligand, Flt3L) FMS-associated tyrosine kinase ligand (FLT3/FLK2 ligand, Ht3L) can be used as a growth factor in hematopoietic stem or progenitor cells or both body. TGFA TGF-α-like cleavage polypeptide, which binds to EGF receptor' and synergizes with TGF-β to promote non-adherent-dependent cell proliferation in softwood gel. Adjuvant adjuvant β-defensin β-defense The prime system is an antimicrobial peptide that is resistant to the innate immune response of many Gram-negative and Gram-positive bacteria, fungi, and viruses. High-mobility group-l(HMGBl) High-mobility group-l(HMGBl) protein is a non-histone chromosomal protein of interleukins, which are local to necrotic cell death and cancer, pathogen invasion, trauma and sepsis. And systemic reactions. S100 phagocytic S100 protein mediator inflammatory response and attraction of inflammatory cells to tissue injury sites are important damage-associated molecular pattern (DAMP) molecules in innate immunity. -170- 201207108 Immunomodulator Function Mannan Glycan (a plant polysaccharide, a polymer of mannose) can be used to produce immune anti-k. BCG (BCG) BCG (BCGX attenuated live mycobacterial species) is used as a vaccine against serious and fatal tuberculosis. Bacterial Lipopolysaccharide (LPS) Bacterial lipopolysaccharide (LPS) is an endotoxin that induces a strong immune response when infected with Gram-negative bacteria. Co-stimulatory molecule (forward) 0X40 ligand The 0X40 ligand (OX40L) belongs to the tumor necrosis factor (ligand) superfamily member 4 (Tnfsf4)' which is expressed on dendritic cells to promote Th2 cell differentiation. 4-1BB Ligand (4-1BBL) 4-1BB Ligand (4-1BBL) belongs to the tumor necrosis factor (ligand) superfamily member 9 (Tnfsf9), which is a type 2 transmembrane glycoprotein and is expressed by activation. 4-1BBL on T lymphocytes induces proliferation of activated peripheral blood T cells and affects activation-induced cell death (AICD). The CD40 CD40 protein belongs to the tumor necrosis factor receptor superfamily 5 and is critical for a wide range of mediators of immune and inflammatory responses, including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. Glucocorticoid-induced tumor necrosis factor family-associated protein (GITR) G1TR can cause potent tumor immunity via T cell stimulation. Administration of anti-GITR monoclonal antibodies (mAbs) elicits strong tumor-specific immunity and clearance of established tumors without inducing significant autoimmune diseases. GITR Ligand (GITRL) GITRL is a ligand for GITR. CD70 CD70 is an interleukin that binds to CD27. It affects T cell activation. It induces proliferation of co-stimulated T cells and promotes the production of cytolytic T cells. LIGHT (HSVgD) Herpesvirus Entering Vector Factor (HVEM) Binding Ligand (HSVgD), also known as p30 or LIGHT, is a member of the TNF family involved in co-stimulation with T cells. PD-L1 (aka CD274) The PD-L1 (aka CD274) protein is expressed in activated monocytes, T cells, and B cells. When treated with IFN-γ, the PD-L1 line in the mononuclear sphere was up-regulated, and PD-L1 in dendritic cells and keratinocytes was also up-regulated when treated with IFN-γ together with other activators. ICOS-L ICOS-L is a T cell-specific cell surface receptor ICOS ligand, which serves as a co-stimulation signal for T cell proliferation and interleukin secretion; it also induces B cell proliferation and differentiation into plasma cells. Costimulatory molecule (negative) Anti-CTLA4 cytotoxic T lymphocyte-associated antigen 4 (CTLA4) is a member of the immunoglobulin superfamily and a costimulatory molecule expressed in activated T cells. 0 Anti-PD-L1 PD- Binding of L1 to the PD-1 receptor on T cells transmits a negative co-stimulatory signal to the cell to prevent the cell from continuing the cell cycle and increasing T cell proliferation. Inhibition of PD-L1 and receptor interactions on T cells with anti-PD-L1 antibodies results in down-regulation of immune responses, termed immune cell failure. -171 - 201207108 Immunomodulator Functional Anti-PD-L2 PD-L2 is associated with T lymphocyte proliferation and co-stimulatory signals required for IFN-γ production in a PDCD1-independent manner, but it is known that this ligand is mainly via PD -1 causes a failure reaction to act. Reverse immunosuppressant (tolerance) TGFR2DN Upon ligand binding, TGFR2 forms a receptor complex consisting of two type II and two type I transmembrane serine/threonine kinases. Type II receptors phosphorylate and activate type I receptors to autophosphorize type I receptors, followed by binding and activation to SMAD transcriptional regulators. Receptor for TGF-β. Deletion of the expected cytoplasmic domain of serine/threonine kinase (nucleotides 1172 to 2036 of TGFpR2 cDNA H2-3FF, available from public databases such as accession number M85079 and amino acid sequence such as accession number AAA61164) All three TGF-βΟ, 2 and 3) dependent gene expression. Anti-TGFp TGFP is a multifunctional peptide that controls the proliferation, differentiation and other functions of many cell types. TGFP synergizes with TGFa on induced transformation. It also acts as a negative autocrine growth factor. Abnormalities in TGFP activation and signaling may lead to apoptosis. Administration of an anti-TGFP antibody prevents renal insufficiency and glomerular sclerosis in db/db mice, which is a type II diabetes model that develops a significant renal disease. Anti-IL10 IL-10 is an interleukin produced by activated Th2 cells, B cells, keratinocytes, mononuclear cells, and macrophages. IL-10 can be used to promote the growth and differentiation of activated human B cells, inhibit the Th1 response to prevent transplant rejection and T cell-mediated autoimmune diseases. Anti-Interleukin Signaling Inhibitor 1 (SOCS1) Interleukin Signaling Inhibitor 1 (S0CS1) is an important inhibitor of interferon gamma signaling, which prevents the possible neonatal effects of this interleukin anti-TGF- a TGF-α-like cleavage polypeptide that binds to the EGF receptor and acts synergistically with TGF-β to promote non-adherent-dependent cellular proliferation in soft rubber gum. Fas contains a cytoplasmic Fas-associated death domain Protein (FADD) FADD is required for Fas and TNF-induced planning cell death (apoptosis) and receptor oligomerization. The biological activity of IL-12 is also well known, including but not limited to differentiation of the initial tau cells into Th1 cells, stimulation of the growth and function of sputum cells, and production of interferon gamma by sputum cells and natural killer (NK) cells. -γ) and tumor necrosis factor-a (TNF-a), reduce IL-4 vector IFN-γ inhibition, enhance NK cell and CD8 + cytotoxicity, cytotoxic activity of lymphocytes, stimulate IL-12R-βΐ and IL The performance of -12R-P2 promotes tumor antigen expression and anti-angiogenic activity via MHC I and II molecules -172-201207108. The phrase "protein having the function of IL-12" comprises a mutant of the wild type IL-12 sequence and a non-IL-12 protein mimicking one or more biological activities of IL-12, wherein the wild type sequence is by one or more The amino acid is changed by addition, deletion or substitution. As used herein, the term "activated" or "activated" refers to any measurable increase in the cellular activity of a gene switch, resulting in the expression of a gene of interest, for example, selected from the group consisting of IL-1, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10R or its subunits DN, IL-15, IL-18, IL-21, IL-23, IL-24, IL-27, GM-CSF, IFN-α, IFN-γ, CCL3 (MIP-1 a) &gt; CCL 5 (R AN TES ), CCL7 (MCP3), XCL1 (lymphocyte chemotactic factor), CXCL1 (MGSA -α) , CCR7 , CCL1 9 (MIP-3b) , CXCL9 ( MIG ) , CXCL 1 0 (IP -1 0), CXCL 1 2 (SDF-1), CC L2 1 (6Ckine), OX40L, 4-1BBL , CD40 'CD70, GITRL, LIGHT, b-defensin, HMGB1, Flt3L, IFN-β, TNF-α, dnFADD, TGF-α, PD-L1 RNAi, PD_L1 antisense oligonucleotide, TGFbRII DN, ICOS- L or S100. As used herein, the term "treating" or "treating" a disease means performing a plan to alleviate the signs or symptoms of the disease, which may include administering one or more drugs or cells engineered in vitro to the mammal. (human or non-human). Therefore, "treatment" should not necessarily be seen as the need to completely alleviate signs or symptoms, not to require a cure, and in particular to include a plan that only has a marginal effect on the individual. As used herein, "immune cells" include dendritic cells, macrophages, neutrophils, obese cells, eosinophils, basophils, natural killer-173-201207108 cells and lymphocytes (eg, B cells and T cells). The terms "dendritic cells" and "DC" are used interchangeably herein. As used herein, the term "therapeutic support cell" (TSC) is a cell which can be modified (eg, transfected, electroporated, etc.) by a vector of the invention to deliver one or more proteins having the function of an immunomodulatory agent and optionally A protein with the function of IL-12 to the tumor microenvironment. The TSC includes, but is not limited to, stem cells, fibroblasts, endothelial cells, and keratinocytes. The term "immune cells engineered in vitro" or "in vitro engineered immune cell population" or "engineered immune cell population" or "immune cells expressing immunomodulators" or "expressing IL" An immune cell of -12 means an immune cell, such as a dendritic cell, conditionally expressing an immunomodulator and/or IL-1 2 under the control of a gene switch, and the gene switch can be activated by an activating ligand. The term "TSC for in vitro engineering" or "over-engineered TSC group" or "engineered TSC group" or "TSC for performance of immunomodulator" or "TSC for IL-12" By means of a conditional expression of an immunomodulator and/or IL-1 therapeutic support cells such as stem cells, fibroblasts, endothelial cells and keratinocytes under the control of a genetic switch, the gene switch can be activated by an activating ligand. As used herein, "modified cells" refers to cells that have been altered by a procedure including, but not limited to, transfection, electroporation, microinjection, transduction, cell fusion, DEAE dextran, calcium phosphate precipitation, and lipids. Transfection (lysosomal fusion). The term "MOI" or "multiplicity of infection" as used herein refers to the average number of adenoviral particles (e.g., recombinant adenovirus or control adenovirus) that stains a single cell in a particular experiment - 174 - 201207108. The term "tumor" as used herein refers to all benign or malignant cell growth and proliferation in vivo or in vitro, whether precancerous or cancerous cells and/or tissues. In another embodiment, the vectors and methods of the invention can be used to treat a disease. In another embodiment, the vectors and methods of the invention can be used to treat cancer. Non-limiting examples of cancers treatable by the present invention include breast cancer, prostate cancer, lymphoma 'skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head and neck cancer , glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck cancer, breast cancer, ovarian cancer, lung cancer, small cell lung cancer, Wilms' tumor, cervical Cancer, sputum, bladder, pancreatic, gastric, colon, prostate, genitourinary, thyroid, esophageal, myeloma, multiple myeloma, adrenal, renal cell carcinoma, endometrial cancer 'Adrenal cortical carcinoma, malignant pancreatic islet tumor, malignant carcinoid tumor, choriocarcinoma, verrucous granuloma, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute Myeloid leukemia, chronic myelogenous leukemia, chronic granulocyte leukemia, acute granulocyte leukemia, hairy cell leukemia, neuroblastoma, horizontal stripes Sarcoma, Kaposi's sarcoma, polycythemia vera, orthostatic thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, mesothelioma -175-201207108 Osteosarcoma, primary macroglobulinemia, and retinoblastoma and similar cancers. In another embodiment, the vectors and methods of the invention can be used to treat metabolically related diseases including, but not limited to, diseases selected from the group consisting of dyslipidemia, atherosclerosis, insulin resistance, diabetes ( For example, Type I diabetes, Type II diabetes, MODY and gestational diabetes), obesity, glucose intolerance, atheroma, hypertension, heart disease (including but not limited to coronary heart disease, stroke, cardiac insufficiency) , coronary insufficiency and hypertension), hyperlipidemia, glucose intolerance, insulin resistance, hyperglycemia, hyperinsulinemia, metabolic syndrome x (or syndrome X, or insulin resistance syndrome, or Raven's (Rea ven's) syndrome, or metabolic cardiovascular risk syndrome), hypertension, chronic fatigue, accelerated aging, degenerative diseases, endocrine deficiency of aging, Gm 1 ganglioside, Morquio B Type disease, Krabbe's disease, Fabry's disease, Gaucher's disease, Tay-Sachs disease, Sandhoff's , fucosidase deficiency, carbohydrate metabolism diseases (such as glycogen storage disease), amino acid metabolism diseases (such as phenylketonuria, maple syrup, glutarate type 1), organic acids Metabolic diseases (eg, black urine), diseases of fatty acid oxidation and mitochondrial metabolism (eg, medium chain thiol dehydrogenase deficiency), porphyrin metabolic diseases (eg acute intermittent porphyria), purine or pyrimidine metabolism Diseases (such as Lesch-Nyhan's disease), steroid metabolic diseases (such as congenital adrenal hyperplasia), and mitochondrial function diseases (such as Kaisers (Kearns_)

Sayre) or peroxisome-functional disease (eg Zellweger's disease). -176- 201207108 In another embodiment, the vectors and methods of the present invention can be used to treat autoimmune diseases including, but not limited to, diseases selected from the group consisting of gastric acid deficiency, autoimmune chronic active hepatitis, acute Diffuse encephalomyelitis, acute hemorrhagic leukoencephalitis, Addison's disease, gammaglobulinemia, gamma globulin deficiency, alopecia areata, amyotrophic lateral sclerosis, joint adhesion spondylitis, anti-GBM/TBM nephritis, antiphospholipid syndrome, anti-synthetase syndrome, arthritis, atopic allergy, atopic dermatitis, aplastic anemia, autoimmune cardiomyopathy, autoimmune hemolytic anemia, Autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune peripheral neuropathy, autoimmune pancreatitis, autoimmune polyendocrine gland disorders I, II, and III Type, autoimmune progesterone dermatitis, autoimmune thrombocytopenic purpura, autoimmune uveitis, Balo disease / balo concentric sclerosis, Bessie (Bechets), Berger's disease, Bickerstaffs encephalitis, B1 au syndrome, large vesicular pemphigus-like disease, Castleman's disease, chronic fatigue immune dysfunction Syndrome, chronic inflammatory demyelinating polyneuropathy, chronic relapsing polymyelitis, Churg-Strauss syndrome, plaque pemphigus, celiac disease, Cogan Syndrome, cold agglutinin, complement component 2 deficiency, cranial arteritis, CREST syndrome, Crohn's disease, Cushing's syndrome, leukocytic vasculitis, Dego's disease , herpes-like dermatitis, dermatomyositis, type 1 diabetes, diffuse cutaneous systemic sclerosis, Dressler's syndrome, discoid lupus erythematosus, eczema, attachment point inflammation -177-201207108 Arthritis, eosinophilic fasciitis, acquired large herpes epidermis, nodular erythema, mixed hypoglycemia, Evan's syndrome, progressive ossifying fibrous dysplasia Fibromyositis, fibrotic alveolitis, gastritis, gastrointestinal pemphigus, giant cell arteritis, Goodpasture's syndrome, Graves' disease, Guillain-Barre Syndrome (GBS), Hashimoto's encephalitis, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schonlein purpura, gestational herpes, Hughes syndrome (or antiphospholipids) Symptoms), hypogammaglobulinemia, spontaneous inflammatory demyelinating disease, spontaneous pulmonary fibrosis, spontaneous thrombocytopenic purpura, IgA nephropathy (or Berger's disease), endosomal muscle Inflammation, demyelinating polyneuropathy, juvenile spontaneous arthritis, juvenile rheumatoid arthritis, Lambert-Eaton myasthenia gravis, leukocyte vasculitis, lichen planus, Sclerosing moss, linear IgA disease (LAD), Lou Gehrig's disease, lupus-like hepatitis, lupus erythematosus, Majeed syndrome, Meniere's disease Microscopic polyangiitis, M i 11 er - F isher syndrome, mixed connective tissue disease, Mucha-Habermann disease, Muckle-Wells Two syndromes, multiple myeloma, myasthenia gravis, myositis, narcolepsy, optic neuromyelitis (also known as Davic's disease), ocular pemphigus, Euclidean 0 rd) thyroiditis, recurrent rheumatism, pa NDAS (autoimmune neuropsychiatric abnormalities in children with streptococcal infection), paraneoplastic cerebellar degeneration, paraneoplastic cerebellar degeneration, Parry R 〇mberg) -178- 201207108 Syndrome, Parsonnage-Turner syndrome, pemphigus, pemphigus vulgaris, pernicious anemia, venous peripheral encephalomyelitis, POEMS syndrome, nodular polyarteritis, rheumatism Polymyalgia, polymyositis, primary biliary cirrhosis, psoriasis, psoriatic arthritis, gangrenous pyoderma, pure red blood cell dysplasia, Rasmussen's encephalitis, Raynaud syndrome, Recurrent polychondritis Reiter's syndrome, posterior peritoneal fibrosis, rheumatoid arthritis, rheumatoid fever, Schmidt syndrome, Schnitzler syndrome, scleritis, spondyloarthropathy, mucin Syndrome, Still's disease, subacute bacterial endocarditis (SBE), Susac's syndrome, Sweet's syndrome, Sydenham's chorea, Sympathetic ophthalmia, Takayasu arteritis, temporal arteritis, Tolosa-Hunt's syndrome, transverse myelitis, ulcerative colitis, undifferentiated connective tissue disease, undifferentiated spondyloarthropathy Vasculitis, Wegener's granulomatosis, Wilson's syndrome, and Wiskott-Aldrich syndrome. In another embodiment, the vectors and methods of the present invention can be used to treat ocular diseases including, but not limited to, diseases selected from the group consisting of open angle glaucoma (eg, primary open angle glaucoma, pigmentation) Glaucoma, exfoliative glaucoma, low intraocular pressure glaucoma), atlas angle closure glaucoma (clinically known as angle-closure glaucoma, narrow-angle glaucoma, pupillary block glaucoma, and ciliary muscle block glaucoma) (eg acute sputum) Angle-locked glaucoma and chronic sacral atresia glaucoma), irido-free glaucoma, congenital glaucoma, juvenile glaucoma, hydrogel-induced glaucoma, neovascular glaucoma-179-201207108 Eyes (eg using vascular endothelial growth factor) VEGF) bait, pigment-derived growth factor (PDGF), endostatin, angiostatin or angiopoietin-1), post-traumatic glaucoma, steroid-induced glaucoma, Sturge-Weber syndrome Glaucoma, uveitis-induced glaucoma, diabetic retinopathy (eg using VEGF decoy, PDGF, endothelium) a carrier of keratin, angiopoietin or angiopoietin-1, and a macular degeneration (for example, a vector comprising VEGF decoy, PDGF, endostatin, angiostatin, angiopoietin-1, ATP-binding Kathy family A) Macular degeneration (eg, using a carrier comprising VEGF decoy, PDGF, endostatin, angiostatin, angiopoietin-1, ATP binding to the Karenia family A member 4), choroidal neovascularization (eg, using a VEGF-containing bait, PDGF, endostatin, angiostatin or angiopoietin-1), vascular leakage and/or retinal edema, bacterial conjunctivitis, fungal conjunctivitis, viral conjunctivitis, uveitis, retroormenal sediment, macula Edema (eg using a carrier containing VEGF decoy, PDGF, endostatin, angiostatin or angiopoietin-1), inflammatory response after intraocular lens implantation, uveal inflammatory syndrome (eg chronic iridocyclitis or chronic) Endophthalmitis), retinal vasculitis (eg, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus, progressive systemic sclerosis, knot Segmental polyarteritis, Wegener's granulomatosis, temporal arteritis, Adamantiades Bechcet's disease, Sjogren's disease, recurrent polychondritis and HLA-B27 correlation Spondylitis, sarcoma, Eles disease, acute retinal necrosis, v〇gt Koyanaki Harada, ocular toxoplasmosis, radiation retinopathy, hyperplasia -180- 201207108 vitreoretinopathy , endophthalmitis, glaucoma (eg, inflammatory glaucoma), optic neuritis, ischemic optic neuropathy (eg, including allotropic NADH dehydrogenase unit 4), thyroid-associated eyelid disease, orbital pseudotumor, pigment Dispersed syndrome (pigmentation glaucoma), scleritis, superficial scleritis choroidal disease (eg "white spot" syndrome including but not limited to acute multiple posterior squamous cells), retinopathy (eg cystic macular edema, central serous choroid) Disease and suspected ocular flora bacteria syndrome (eg, a carrier containing glial fine cell-derived neurotrophic factor peripheral-2), retinal vascular disease (eg, diabetes) Disease retinopathy, Coat's disease and retinal artery aneurysm), retinal artery occlusion, retinal vein occlusion, retinopathy of prematurity, retinitis pigmentosa (eg vector containing retinal pigment-specific 65 kDa protein), family exudation Invasive vitreoretinopathy (FEVR), spontaneous polypoid choroidal vascular disease, preretinal macular membrane or cataract plaque Limited to blood diseases selected from the group consisting of anemia, hemorrhage, and coagulopathy (eg, disseminated intravascular coagulation (DIC), hemophilia, Henoch-Schonlein's purpura, hereditary hemorrhagic microvascular dilatation syndrome) , thrombocytopenia (ITP, TTP), thrombotic dysfunction, Von Willebrand's disease, leukemia (eg acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia) , lymphoma (eg Hodgkin's lymphoma 'non-Hodgkin's lymph ), myeloproliferative diseases (such as myelofibrosis, polycythemia vera, thrombocytopenia -181 - 201207108 (thrombocythemia)), purple cell disease (such as macroglobulinemia, monoclonal globulin disease of unknown clinical significance, Multiple myeloma), spleen disease, white blood cell disease (eg basophilic abnormalities, eosinophilic abnormalities, lymphopenia, mononuclear abnormalities, neutrophils, neutropenia), thrombosis, Deep vein thrombosis (DVT), hemochromatosis, menorrhagia, sickle cell disease or thalassemia. In another embodiment, the vectors and methods of the invention can be used to treat neurological diseases including, but not limited to, neurological diseases selected from the group consisting of Gaucher's disease, Parkinson's disease, Aziz. Alzheimer's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Huntington's disease, Fredrich's ataxia, mild cognitive impairment, Cerebral amyloid angiopathy, Parkinson's disease, Lewy's body disease, frontotemporal dementia (FTD), multiple systemic degeneration (MSA), progressive supranuclear nerve palsy, mobile disease (including ataxia) , cerebral palsy, dance refers to rickets, muscle insufficiency, Toray's disease, nuclear jaundice), trembling disease, white matter dystrophy (including adrenal white matter dystrophy, metachromatic white matter dystrophy, white matter spongy Degeneration, Alexander, Pelizaeus-Merzbacher's disease, neuronal waxy lipofuscinosis, ataxia microvascular dilatation syndrome, Rett syndrome, alpha-synucleinopathy For example, Louise Lewy) Body disease, multiple systemic degenerative disease, Hallervorden-Spatz's disease or frontotemporal dementia, Niemann-Pick's type C disease (NPCD), spinocerebellar atrophy Type 1, 2, and 3 or dentate nucleus red globus pallidus atrophy (DRLPA). -182- 201207108 In another embodiment, the vectors and methods of the invention can be used to treat pulmonary diseases including, but not limited to, pulmonary diseases selected from the group consisting of asthma, pulmonary insufficiency, bronchitis, COPD (chronic obstruction) Pulmonary disease), emphysema, lung cancer, mesothelioma, pneumonia, asbestosis, sputum, sputum, acute invasive sputum, bronchiectasis, obstructive bronchiolitis, organizing pneumonia (B OOP) ), eosinophilic pneumonia, necrotizing pneumonia, pleural effusion, pneumoconiosis, pneumothorax, pulmonary actinomycosis, alveolar proteinosis, pulmonary anthrax, pulmonary arteriovenous malformation, pulmonary fibrosis, pulmonary embolism, lung cell growth Syndrome X (eosinophilic granuloma), pulmonary hypertension, pulmonary edema, pulmonary hemorrhage, pulmonary soil filariasis, tuberculosis, pulmonary venous occlusive disease, rheumatoid lung disease, sarcoma-like disease, radiofibrosis, hypersensitive pneumonia , acute respiratory distress syndrome (ARDS), infant respiratory distress syndrome, spontaneous pulmonary fibrosis, spontaneous interstitial pneumonia, lymphangioleic smooth muscle hyperplasia, lung Langerhans' Increase organizational goals disease, alveolar proteinosis, nose treasure, tonsillitis, otitis media, pharyngitis, laryngitis, pulmonary hamartoma, free lung, congenital cystic adenomatous malformation (C CAM) or cystic fibrosis. In another embodiment, the vectors and methods of the invention can be used to treat rheumatic diseases including, but not limited to, rheumatic diseases selected from the group consisting of systemic lupus erythematosus, dermatomyositis, scleroderma, system Necrotizing arteritis, epidermal necrotizing phlebitis, rheumatoid arthritis, Sjogren's syndrome, Raynaud's syndrome, Reiter's syndrome, arthritis, psoriasis joints Inflammation, seronegative spondyloarthropathy, Sjogren's syndrome, systemic sclerosis, dermatomyositis/polymyositis, mixed connective tissue disease or joint adhesion spine-183-201207108 inflammation. In another embodiment, the vectors and methods of the invention can be used to treat infectious diseases including, but not limited to, infectious diseases selected from the group consisting of fungal diseases such as dermatophytosis (eg, trichophytonosis, Money or round sputum infection), Hong Kong foot, paronychia, discoloration pityriasis, red mites, rash, fungal diaper rash, Candida vulvitis, Candida balanitis, otitis externa, candidiasis (skin and mucous membrane Skin), chronic mucosal candidiasis (eg, thrush and vaginal candidiasis), cryptococcosis, dermatophysosis] trichosporosis, sputum, penicillosis, sputum, zygomycete, spore Filariasis, chromogenic fungal disease, coccidiosis, tissue aspergillosis, blastomycosis, paracoccus, pseudo-endococcosis, foot edema, fungal keratitis, acne, pneumocystis And true septicemia, Acinetobacter infection, actinomycosis, African sleeping sickness, AID S (acquired immunodeficiency syndrome), amebiasis, echinococcosis, anthrax, hemolytic bacterium, Argentine hemorrhagic fever, mites Disease, sputum disease, astrovirus , coccidiosis, Bacillus licheniformis infection, bacterial pneumonia, bacterial vaginosis (BV), bacillus infection, ciliate disease, Baylisian elegans (Baylisascaris) infection, BK virus infection, black hair shaft disease, human Blastocystis infection, B. burgdorferi infection, botulism (and infant botulism), Brazilian hemorrhagic fever, brucellosis, Burk ο 1 deria infection, bulu (B uru 1 i) ulcer, calicivirus infection (Norovirus and Sapovirus), candidiasis, cat scratching, cellulitis, Chagas disease ( Chagas disease), soft chancre, varicella, chlamydial infection, cholera, pigmented blastomycosis, liver fluke, Clostridium difficile, cocci - 184 - 201207108 bacteria, Colorado heat (CTF ), common cold (acute viral nasopharyngitis, acute rhinitis), Creutzfeldt-Jakob disease (CJD), cryptococcosis, cryptosporidiosis, larval larvae migration (CLM), dengue fever, brittle dinuclear Miba disease, diphtheria , schistosomiasis, schistosomiasis, Dracunculiasis, Ebola hemorrhagic fever, echinococcosis

(Echinococcosis), Ehrlichiosis, Entobiasis (Aphid infection), Enterococci (E nter 〇coccus) infection, Enterovirus infection, Epidemic typhus (Epidemic Typhus) 'Erythema infectiosum, Exanthem subitum, Fasciolopsiasis, Fasciolosis, Fatal Familial Insomnia (FFI), Filariasis ), Fusobacterium infection, gas gangrene (Cymenthus muscle necrosis), Geotrichosis, Gerstmann-Straussler-Scheinker syndrome (GSS), Piriformis (Giardiasis) ), G1 an ders, Gnathostomiasis, Gonorrhea, Granuloma inguinale (Donovanosis), Group A streptococcal infection, Group B streptococci Infection, Haemophilus influenzae, Hand, Foot and Mouth Disease (HFMD), Hantavirus (HPS), Helicobacter pylori infection, hemolytic uremic syndrome (HUS), hemorrhagic fever with renal syndrome (HFRS), hepatitis A, hepatitis B, hepatitis B, hepatitis D, hepatitis E, herpes simplex, Histoplasmosis, hookworm infection, human Bocavirus infection, human ewingii ehrlichiosis, human granule ball-185- 201207108 granulocytic anaplasmosis (HGA), human granule porphyria (HGA) ), human mononuclear arsenic, human papillomavirus (HPV) infection, human parainfluenza virus infection, membrane-like ascariasis (Hymenolepiasis), EB virus infectious mononucleosis (£?316111) -Barr Virus Infectious Mononucleosis) (Mono), influenza (flu), Isosporiasis, Kawasaki disease, keratitis, Kingella kingae infection, Kuru disease ), Lassa fever, Legionellosis (Veterans' Disease), Legionnaires' Disease (Pontiac fever), Leishmaniasis, Leprosy ), Leptospirosis, Li Listeriosis, Lyme disease (Lymbibiosis), lymphatic filariasis (Elephantiasis), Lymphocytic choriomeningitis, Malaria, Marburg hemorrhagic fever (MHF), hemorrhagic 'Melioidosis' (Whitmore's disease), meningitis, meningococcal disease, mitochondrial disease (Metagonimiasis), Microsporidiosis, infectious soft sputum (MC), adenitis, Murine typhus (Endemic typhus), Mycoplasma pneumonia, foot bacteria Mycetoma, Myiasis, Neonatal Conjunctivitis (Newborn Ophthalmitis), (New) Variant Creutzfeldt-Jakob Disease CJD, nvCJD), Soil Nocardiosis, Plate Helicobacteriasis (river blindness), paracoccidioidomycosis (Southern bud disease -186-201207108 (blastomycosis)), paragonimiasis (Paragonimiasis), Pasteurellosis (P "asteurellosis", cephalosporosis (Pediculosis capitis), cephalosporin (Pediculosis corporis), edulis (Pediculosis pubis), pelvic inflammatory disease (PID), whooping cough (Pertussis) (Whooping cough), Plague, pneumococcal infection, Pneum〇cystis pneumonia (PCP), pneumonia, poliomyelitis (Poliomyelitis), poliomyelitis, Platts Prevotella infection, primary amoebic meningoencephalitis (PAM), progressive multifocal leukoencephalopathy, Psittacosis, Q fever, rabies, Rat-bite fever, Respiratory fused cell viral infection, Rhinosporidiosis, Rhinovirus infection, Rickettsial infection, Rickettsialpox, Rift Valley feverKRVF, Rocky mountain spotted fever (RMSF), Rotavirus infection, Rubella, Salmonellosis, SARS (Severe Acute Respiratory Syndrome), 疥(Scabies), Schistosomiasis, sepsis, Shigellosis (Bacillus dysentery), Shingles (Herpes zoster), Smallopox (Variola), Sporotricosis ), Staphylococcus aureus food poisoning, Staphylococcus aureus infection, Strongyloidiasis, Syphilis, Taeniasis, tetanus (Lockj aw), Tinea barbae (Barber's itch) ), Tinea capitis (the scalp money), Tinea corporis (-187- 201207108 body money 癣), Tinea cruris (Jock itch), hand lice (Tinea manuum) ( Hand money), Tinea nUra, Tinea ungumm (〇nychomycosis), Tinea versicolor (Pityriasis versicolor), Ascaria ( Toxocariasis), visceral larval migration (VLM), Toxoplasmosis, Trichinellosis, Trichomoniasis, Trichuriasis (whipping), tuberculosis, rabbit fever ( Tu Laremia), Ureaplasma urealyticum infection, Venezuelan equine encephalitis, Venezuelan hemorrhagic fever, viral pneumonia, West Nile Fever, white hair dry sarcoidosis (white Piedra) (Tinea blanca), Y ersiniapseudotubercu 1 osis infection, Yersiniosis, yellow fever or zygomycosis (Zygomycosis) In another embodiment, the vectors and methods of the invention can be used in mammals to treat one or more diseases. In one aspect, the mammal is a human. In another aspect, the mammal is a non-human animal. A variety of diseases that can be treated using the disclosure of the present invention can be readily considered. These diseases include, but are not limited to, chronic kidney disease, osteoarthritis, tumors, viral upper respiratory tract infections, cat plasma cell stomatitis, cat eosinophilic granuloma, feline leukemia infection, canine distemper infection, systemic fungal infection, myocardium Disease, mucopolysaccharidosis type V11 or infectious disease. In one aspect, the disease treated by the present invention is an infectious disease of an animal-188-201207108 disease, including but not limited to bovine respiratory disease, porcine respiratory disease, avian influenza, avian infectious bronchitis, and cattle. Spongiform encephalopathy, canine leishmaniasis, chronic wasting disease, human immunodeficiency virus (HIV), hepatitis, hepatitis B, hepatitis B, hepatitis C, classical swine fever, echinococcosis, endemic pneumonia, FIP, foot-and-mouth disease, lung adenoma, Maedi-Visna disease, mastitis of animals, canine microsporosis, infectious aphthous (animal disease), small ruminant disease, pox, parrot喙feel, rabies, Mediterranean fever ( brucellosis) or Bang's disease or wave heat, malt heat, infectious abortion, epidemic abortion, salmonella food poisoning, intestinal paratyphoid, bacillary Dysentery, pseudotuberculosis, plague, plague fever, tuberculosis, vibriosis, circle disease, Weil's disease (Leptospirosis) or Leptospira fever, hemorrhagic jaundice (hemorrhagic scutellaria), milk Worker fever (Hart's cochlear), regenerative heat, regenerative heat, spiral heat, vagrant fever, famine fever, Lyme arthritis, Bannworth's syndrome (Lyme disease), cerebral meninges Neuritis, chronic migratory erythema, vibrosis, Escherichia coli, E. coli toxemia, ferrets, swine intestinal edema, intestinal paratyphoid, staphylococcal gastrointestinal poisoning, staphylococcal gastroenteritis, canine coronavirus ( CCV) or canine prion enteritis, feline infectious peritonitis virus, infectious gastroenteritis (TGE) virus, Hagerman's red mouth disease (ERMD), infectious hematopoietic necrosis (IHN), A. porcine Hemobacteria) Pleuropneumonia, Hansen's disease, Streptomyces, Sheep fungal dermatitis, False sinus, Whitmore's disease, Francis' disease, Deer fever, Rabbit Fever, O'Hara's disease, Streptomyces fever, Haverhill-189- 201207108 (Haverhill) fever, epidemic joint erythema, rat bite heat 'ship or transport heat, hemorrhagic sepsis, parrot Disease, parrot fever, chlamydia , North American blastomycosis, Chicago disease, Gilchrist's disease, cat scratching heat, benign lymphoid reticuloendothelial cells, benign non-bacterial lymphadenitis, bacillary aneurysm disease, bacillary purpura hepatitis, Q fever, Balkan influenza, Balkan grippe, slaughterhouse fever, sputum heat, lung rickettsial disease, American typhus typhus, sputum typhus fever, vesicular ricketts Secondary disease, Royal Botanic Garden spot fever, sputum typhus fever, endemic typhus fever, urban typhus, money sputum, dermatophytosis, sputum, sputum bacillus, microsporosis, stocks癣, Hong Kong foot, S. skrjab, dimorphic fungus, cryptococcosis and histoplasmosis, benign epidermal monkeypox, BEMP, simian herpes virus, simian B virus disease, type C lethargy encephalitis, yellow Fever, black spitting, Hanta virus pulmonary syndrome, Korean hemorrhagic fever, epidemic nephropathy, epidemic hemorrhagic fever, hemorrhagic nephritis, lymphocytic choriomeningitis, Venezuelan equine encephalitis, California encephalitis/Lackos brain Inflammation, African hemorrhagic fever, green monkey disease, fear of water, rabies, infectious hepatitis, epidemic hepatitis, epidemic jaundice, rash, measles, swine flu, horse flu, chicken cockroach, Newcastle disease, pear larvae, bow Insect disease, African sleep disease, Gambia trypanosomiasis, Rhodesia trypanosomiasis, Chagas disease, Chagas-Massachosis, Chagas disease, E. histolytica, Peony Dysentery, cryptosporidiosis, pear-shaped flagellate, cutaneous leishmaniasis; gum worker ulcer, nasopharyngeal mucosal leishmaniasis, pianbols, uta and bub a (American), oriental sore, oriental scorpion (old world) , Baghdad, New Delhi, Bauru ulcer, organ leishmaniasis: kala-azar, microsporidia, -190- 201207108 Gastric nematode, trichinosis, angiostrongosis, eosinophilic meningitis or meningoencephalitis (Guangdong live blood nematode), abdominal avascular nematode disease (Costaric sinensis), hookworm disease, scutellaria, hookworm, capillary nematode, malaria filariasis, tsutsugamushi, nodular nematode Disease, roundworm, roundworm, chicken Insect, schistosomiasis, schizophrenia, echinococcosis, echinococcosis, Echinococcus granulosis, cystic echinococcosis, mites infection, Schistosoma and Similar diseases. It is also considered to treat malignant diseases caused by infectious pathogens. Examples of such diseases include, but are not limited to, osteosarcoma, leukemia, lymphoma, Burkitt's lymphoma caused by EBV, Lloyd's sarcoma caused by Rous retrovirus, and type 8 blister Kaposi sarcoma caused by rash virus, adult T cell leukemia caused by HTLV-I retrovirus or cell-like leukemia caused by HTLV-II and many other tumors and leukemia caused by infectious agents and viruses.

In one embodiment, the one or more proteins for treating one or more of the diseases include, but are not limited to, erythropoietin, ghrelin, osteoproteger, RANKL, RANKL decoy, TNF-α antagonist, IL-1 Antagonists 'G-CSF, GM-CSF, IFN-a, IFN-γ, angiostatin, endostatin, TNF-a, PP1DCY-LSRLOC, β-urtase and IL-12. In another embodiment, one or more proteins of the invention include, but are not limited to, IL-1, IL-2, IL-12, IL-3, IL-4 'IL-5, IL-7, IL-8 , IL-9, IL10RDN or subunits, IL-15, IL-18, IL-21, IL-23, IL-24, IL-27, GM-CSF, IFN-a, IFN-γ, IFN- a 1, IFN-191 - 201207108 α 2, IL-15-Ra, CCL3 (MIP-la), CCL5 (RANTES), CCL7 (MCP3), XCL1 (lymphocyte chemotactic factor), CXCL1 (MGSA-α), CCR7, CCL19 (MIP-3b), CXCL9 (MIG), CXCL10 (IP-10), CXCL 1 2 (SDF-1), CCL 2 1 (6 C kine), OX40L, 4-1BBL, CD40, CD70, GITRL , LIGHT, b-defensin, HMGB1, Flt3L, IFN-β, TNF-α, dnFADD, BCG 'TGF-α, PD-L1 RNAi, PD-L1 antisense oligonucleotide, TGFbRII DN, ICOS-L and S100 = In one embodiment, the vector is administered to an adenoviral vector that is afflicted with one or more mammals of the disclosed disease. In one embodiment, the vector comprises a polynucleotide encoding a gene switch. In one aspect, the gene switch is an EcR-substrate gene switch. In another embodiment, the polynucleotide encoding the gene switch comprises a first transcription factor sequence under the control of a first promoter and a second transcription factor sequence under the control of a second promoter, wherein the first transcription is The factor sequence and the protein encoded by the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. In one aspect, the system is diterpene. In another aspect, the ligand is selected from the group consisting of RG-115819, RG- 1 1 5932 or RG- 1 1 5 83 0. In yet another aspect, the system is amidoxime or oxadiazoline. In another embodiment, the invention can be used in a mammal to treat one or more lysosomal storage diseases. In one aspect, the mammal is a human. In another aspect, the mammal is a non-human animal. Examples of lysosomal storage diseases that can be treated by the present invention include, but are not limited to, Pompe's disease/hepatic glucose storage type II, Gaucher's disease (type I, type II, type III) , Fa-192- 201207108 Bab (Fabry), mucopolysaccharidosis type II (Hunter's syndrome), mucopolysaccharidosis type VI (Maroteaux-Lamy's syndrome), mucopolysaccharide Type I, metachromatic white matter dystrophy, neuronal waxy lipofuscinosis or CLN6 disease (atypical late infant, late-onset variant, early juvenile, Finnish variant late infant CLN5, Zhan - (Jansky-Bielschowsky) second disease/late infant type CLN2/TPP1 disease, Kufs/adult NCL/CLN4 disease, northern epilepsy/variant late infant type CLN8, Santavuori-Haltia 2nd / infant type 〇 CLN1/PPT disease, β-mannose disease), Batten-Spielmeyer-Vogt 2nd/juvenile NCL/CLN3 disease, type A St. Philippe (Sanfilippo) syndrome, Type B St. Philippine Syndrome, Type C St. Philip's Syndrome, Type D St. Philip's Syndrome, and MPSI Hurler's Symptoms , Niemann-Pick disease (type A, type B, type C, type D), activator deficiency / GM2 ganglioside, alpha-mannoseosis, aspartame glucosamine , cholesteryl ester storage disease, chronic hexosaminidase deficiency, cystine U disease, Danon's disease, Farber's disease, fucosidase deficiency, half Lactose sialidase deficiency disease (Goldberg syndrome), GM1 ganglioside (infant type, late infant/juvenile type, adult type/chronic), I-cell disease/viscosity type II, infancy Free sialic acid storage disease / ISSD, juvenile hexosaminidase A deficiency, Krabbe's disease (infancy, late onset), mucopolysaccharidosis (Hurler's multiple nutrition) Adverse Disorders / Mucins Type IIIA, Scheie's Disease, MPS Type I Hale Shay Syndrome, Morquio's Type A/MPS IVA Type, Morki's Disease B Type / MPS Type IVB, MPS Type IX - 193 - 201207108 Vitalonic acid deficiency, Sly (MPS type VII), viscid type i / sialic acidosis Mucoidosis type IIIC, viscid type IV), multiple sulphate lipase deficiency, dense osteogenesis imperfecta, Sandhoff's disease/adult-occurring type/GM2 ganglioside, Shandehu Furu disease / GM2 gangliosides - infant type, Sandhoff's disease / G Μ 2 gangliosides - juvenile type, Schindler's disease, Salla's disease, Infant sialic acid storage disease, T ay - S achs / G Μ 2 gangliosides, Wolm an's disease, aspartame glucosamine and sphingolipid activation Proteinogen. It should be understood that the Sanfilippo syndrome type A is synonymous with type A St. Philippine syndrome/MPS I1IA, the type B St. Philippine syndrome is synonymous with type B St. Philip syndrome/MPS IIIB, the type C St. Philip syndrome and the type C St. Philip Syndrome/MPS II 1C synonymous, D-type St. Philip syndrome group is synonymous with D-type St. Philip syndrome/MPS I1ID. In one embodiment, the one or more proteins expressed by the vector of the present invention for use in the treatment of one or more lysosomal storage diseases include, but are not limited to, a-galactosidase A, arylsulfatase A , a-glucosidase, b-glucosidase, glucocerebrosidase, CLN6 protein, juvenile-related CLN3, N-thioglucosamine hydrolase (SGSH), aN-acetylglucosamine, acetamyl coenzyme A-glucosamine glycosyltransferase, N-acetylglucosamine-6-sulfate lipase, aL-iduronidase, arylsulfatase B, acid sphingomyelinase and iduroxaldehyde Acid sulfate lipase. In one embodiment, the vector is administered to an adenoviral vector that is afflicted with one or more mammals that reveal the lysosomal storage disease. In one embodiment, -194-201207108, the vector comprises a polynucleotide encoding a gene switch. In one aspect, the gene is associated with an EcR-substrate gene switch. In another embodiment, the polynucleotide encoding the gene switch comprises a first transcription factor sequence under the control of a first promoter and a second transcription factor sequence under the control of a second promoter, wherein the first transcription is The factor sequence and the protein encoded by the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. In one aspect, the system is diterpene. In another aspect, the system is selected from the group consisting of RG-115819, RG- 1 1 5932 or RG- 1 1 5 83 0. In yet another aspect, the system is amidoxime or oxadiazoline. In another embodiment, the invention can be used in a mammal to treat one or more liver diseases. In one aspect, the mammal is a human. In another aspect, the mammal is a non-human animal. In one aspect, the liver disease is hepatitis B. In another aspect, the liver disease is hepatitis C. In one embodiment, the protein represented by the vector of the present invention is IFN-α. In another embodiment, the protein expressed by the vector of the present invention for use in the treatment of one or more of the liver diseases described above comprises copper blue plasma protein. A non-limiting example of a human liver chimeric mouse model for hepatitis B virus infection and treatment is disclosed in Bissig, K. D.  Et al. , J.  Clin.  Investigation 1 20: 924 (20 1 0). A non-limiting example of another human hepatocyte model is the humanized mouse system of YeCUriSTMl] (Portland, OR). A non-limiting example of an encephalitis model for assessing antiviral/anti-infective therapies is disclosed in O’Brien, L.  Et al. , J.  General Virology 90: 874-882 (2009). •195- 201207108 A non-limiting example of a flu model for assessing antiviral/anti-infective treatments is disclosed in Beilharz, M. W.  Et al. , Biochemical Biophysical Research Communications 3 5 5: 740-744 (2007) and Koerner, I.  Et al. , J.  Virology 8 1: 2 0 2 5-2030 (2007) ° In one embodiment, the vector is administered to an adenoviral vector that is afflicted with one or more mammals that exhibit liver disease. In another embodiment, the vector is not an adenoviral vector. In another embodiment, the vector is a plastid. In one embodiment, the vector comprises a polynucleotide encoding a gene switch. In one aspect, the gene is associated with an EcR-substrate gene switch. In another embodiment, the polynucleotide encoding the gene switch comprises a first transcription factor sequence under the control of a first promoter and a second transcription factor sequence under the control of a second promoter, wherein the first transcription is The factor sequence and the protein encoded by the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. In one aspect, the system is dimeric. In another aspect, the system is selected from the group consisting of RG-1 15819, RG-115932, or RG-115830. In yet another aspect, the system is amidoxime or oxadiazoline. The present invention provides an engineered cell such as an immune cell and TsC to conditionally express a protein having an immunomodulatory agent and optionally a function of IL_1 2, and the protein has a treatment for treating cancer or a tumor or both. Sexual use and / or application. Immunogens and TSCs which are conditionally expressed as proteins having an immunomodulatory agent and optionally a function of IL-1 2 by in vitro engineering are safer than constitutively produced protein systems. In addition, the ability to control the immunomodulator and optionally the performance of IL-1 2 -196-201207108 provides better control of the treatment effect. Therefore, the in vitro engineered immune cells and TSC can be formulated into a pharmaceutical composition as a therapeutic agent for treating cancer or tumor of a human or non-human organism. Alternatively, an in vitro engineered immune cell population, a TSC population, or a subgroup thereof can be used as a vehicle to conditionally deliver an immunomodulatory agent and optionally produce IL-1 2 protein to human or non-human A specific area of the body of the organism (normal tissue, cancer, or tumor). The immune cell can be an autologous or non-autologous dendritic cell. The dendritic cells can be isolated from the bone marrow or peripheral blood circulation. In human patients, dendritic cell populations can be isolated by leukocyte separation, in which white blood cell components are separated and removed, and other blood components are reintroduced into the patient. In another embodiment, the dendritic cells can be transfected with a human hematopoietic stem cell by a vector of the present invention, the vector exhibiting a protein having the function of an immunomodulator and a protein optionally having the function of IL-12. Then, the transfected stem cells are differentiated to produce dendritic cells for preparation. See U.S. Patent No. 6,73,0414. In one embodiment, the invention provides a nucleic acid adenoviral vector comprising a gene switch, wherein the VP16-RXR and GaI4-EcR coding sequences separated by an EMCV internal ribosome entry site (IRES) sequence are inserted by human ubiquitin C Promoter-controlled adenoviral shuttle vector. For example, the coding sequences for the subunits p 4 0 and p35 of IL 1 2 separated by the IRES sequence and placed under the control of a synthetic inducible promoter are inserted upstream of the ubiquitin C promoter. In another example, the coding sequence for TNF-[alpha] placed under the control of a synthetic inducible promoter is inserted upstream of the ubiquitin C promoter. In another embodiment, the invention provides a shuttle vector carrying two fusion proteins and a transcription unit (VP16-RXR and Gal4-EcR) of an inducible IL-12 or TNF-α subunit of -197-201207108 With E. coli (E.  Coli) Recombination of adenoviral skeleton (A d E a s y 1) in B J 5 1 8 3 cells. Carry rAd after confirming the recombinant clone. The RheoIL12 genome system was grown and purified from XL 10-Gold cells. After digestion of the plastid backbone, it was packaged by transfection into HEK 293 cells or CHO cells. Purification of the vector to increase the concentration can be accomplished by any suitable method, such as by density gradient purification (e.g., chlorinated milling (CsCl)) or by chromatographic techniques (e.g., column or batch chromatography). For example, the vector of the present invention can be subjected to two or three CsCl density gradient purification steps. A vector, such as a replication-deficient adenoviral vector, is deliberately purified from cells infected with the replication-defective adenoviral vector, the method comprising dissolving the adenovirus-infected cells, and adding the lysate to the chromatography resin, from the layer The resin is eluted to elute the adenovirus, and the component containing the adenovirus is collected. In a particular embodiment, the resulting original viral stock solution is amplified by reinfection of HEK 293 cells or CHO cells and purified by CsCl density gradient centrifugation. In one embodiment, the immunomodulatory agent, such as the TNF-ct and/or IL-12 gene, is a wild-type gene sequence. In another embodiment, the immunomodulatory agent, e.g., the TNF-[alpha] and/or IL-12 gene, is a modified gene sequence, such as a chimeric sequence or a sequence modified to use a preferred codon. In one embodiment, the immunomodulatory agent, such as the TNF-a and/or IL-12 gene, is a human wild type sequence. In another embodiment, the sequence is at least 85% identical to the -198-201207108 wild-type human sequence, e.g., at least 90%, 95%, or 99% identical to the wild-type human sequence. In other embodiments, the gene sequence encodes a human polypeptide. In another embodiment, the gene encodes a polypeptide that is at least 85 % identical to the wild-type human polypeptide, e.g., at least 90%, 95%, or 99% identical to the wild-type human polypeptide. In one embodiment, the IL-12 gene is a wild type mouse IL-12 sequence. In another embodiment, the sequence is at least 85% identical to wild type mouse IL-1 2, for example at least 90%, 95% or 99% identical to wild type mouse IL-12. . In other embodiments, the IL-12 gene sequence encodes a mouse IL-1 2 polypeptide. In another embodiment, the gene encodes a polypeptide that is at least 85% identical to wild-type mouse IL-12, for example at least 90%, 95% or 9 with wild-type mouse IL-12. 9% consistency. DCs can be isolated from the bone marrow of humans, mice or other mammals. Dendritic cells can be isolated from the blood of humans, mice or other mammals. In human patients, dendritic cell populations can be isolated by leukocyte isolation known in the art, in which white blood cell components are separated and removed, and other blood components are reintroduced into the patient. In one embodiment, the DC system is derived from murine bone marrow as previously described (Tatsumi et al. , 2003). Briefly, wild-type or EGFP Tg mouse bone marrow (BM) was conditioned medium supplemented with 1 000 units/ml recombinant mouse granule/macrophage colony-stimulating factor and recombinant mlL-4 (Peprotech, Rocky Hill, NJ). (CM), cultured in a humidified '5% C02 incubator at 37 °C for 7 days. The CDllc+ DC is then separated using a specific MACSTM bead, according to the manufacturer's instructions (Miltenyi Biotec, Auburn, CA). The CD 11 c+ DC produced in this manner is morphologically pure &gt; 95%, and together exhibits -199-201207108 CD11b, CD40, CD80 and type I and II MHC antigens. An embodiment of the present invention provides an engineered immune cell and TSC that conditionally express a protein having an immunomodulatory agent and optionally a function of IL-1 2, and the engineered immune cell and TSC are suitable as a treatment Therapeutic application of gene therapy for cancer or tumor of human or non-human organisms or both. In one embodiment, the invention provides engineered immune cells and TSCs comprising a gene switch. In another embodiment, the invention provides engineered immune cells and TSCs comprising to a portion of an ecdysone receptor. In another embodiment, the invention provides an engineered immune cell and TSC comprising a ecdysone receptor basal gene switch. In another embodiment, the invention provides engineered immune cells and TSCs comprising RheoSwitch. In another embodiment, the invention provides a kit comprising engineered immune cells and a TSC and a ligand comprising a gene switch and the ligand modulating the gene switch. In another embodiment, the kit further comprises a dimercaptopurine ligand. In another embodiment, the kit further comprises RG-1 1 5 83 0 or RG-1 1 5932. In one embodiment, the invention provides an engineered population of immune cells and a TSC population. In one embodiment, the DC day of culture day is treated with a recombinant adenovirus encoding an immunomodulatory agent and/or IL-12 controlled by a constitutive or inducible promoter, or a plurality of infections ( M0I) was infected with the mock mock adenovirus vector (rAdv^5). After 48 hours, the infected DCs were collected for analysis of the phenotype and the detection limit was 62. 5 picograms per milliliter of specific ELISA kit (BD-PharMingen, San Diego, CA) for analysis of immunity -200-201207108 Modulator and/or IL-12 production. In another embodiment, the invention provides an in vitro engineered immune cell population and a TSC population, the cell population and the TSC population comprising a protein capable of conditionally exhibiting an immunomodulatory agent and/or IL-12 function The vector of the gene switch is, for example, a DNA vector, and further comprises an activating ligand. In another embodiment, the invention provides a method of treating cancer, such as melanoma or glioma, by administering an engineered DC to a patient, and then administering an activating ligand such as RG-115819, RG -115830 or RG-1 1 5 93 2 to the patient. In certain embodiments, the invention relates to a method of treating cancer, such as melanoma or prostate cancer, comprising administering an adenovirus comprising a polynucleotide to conditionally express an immunomodulatory agent, such as TNF-[alpha], and administering the antibody. Ligand. The patient may be a human or animal suffering from cancer. The methods and products, engineered cells, kits and ligands have applications in human therapy and veterinary animal therapy. Accordingly, such products and methods are contemplated for human and veterinary animal purposes. Therefore, in one embodiment, the polynucleotide exhibiting an immunomodulatory agent such as TNF-α and an activating ligand are co-administered to a patient with cancer. The activating ligand is typically administered within a few days, for example, before and after administration of the polynucleotide. Administration of the activating ligand can be reduced or eliminated to alleviate adverse effects if systemic toxicity by an immunomodulatory agent such as TNF-[alpha] occurs. In another embodiment, a polynucleotide exhibiting an immunomodulatory agent, such as TN F-α, and an activating ligand are co-administered to a patient suffering from one or more lysosomal storage diseases or one or more liver diseases. The activating ligand is typically administered within a few days&apos;, e.g., before and after administration of the polynucleotide. In the case of systemic toxicity -201 201207108, administration of the activating ligand can be reduced or eliminated to alleviate adverse reactions. In certain embodiments, the invention provides a method of reducing tumor size, comprising administering an adenoviral vector and administering an activating ligand comprising a polynucleotide conditioned to exhibit an immunomodulatory agent, such as TNF-[alpha] . The invention also provides a method of preventing tumorigenesis comprising administering an adenoviral vector and administering an activating ligand comprising a polynucleotide that conditionally expresses an immunomodulatory agent, such as TNF-[alpha]. In some embodiments, the invention provides a method of reducing or ameliorating the symptoms of one or more neoplastic diseases, comprising administering an adenoviral vector and administering an activating ligand comprising a conditionally indicative immunomodulatory agent, such as TNF - alpha polynucleotide. In particular, a composition comprising a conditionally expressing an immunomodulatory agent, such as an adenoviral vector, reduces, prevents or improves the systemic toxicity of the individual being treated as compared to a constitutively representative carrier of the immunomodulatory agent. In certain embodiments, the invention provides a method of treating one or more diseases or one or more lysosomal storage diseases or one or more hepatic sputum diseases in a mammal. The method comprises administering an adenoviral vector and administering the same A ligand comprising a polynucleotide that conditionally expresses one or more proteins. In some embodiments, the invention provides a method for reducing or ameliorating one or more diseases or one or more lysosomal storage diseases or one or more symptoms of one or more liver diseases in a mammal. Adenoviral vector and administration of an activating ligand. The vector comprises a polynucleotide which conditionally expresses an immunomodulator such as TNF-[alpha]. Protein basal tags reduce or eliminate the need for highly specific post-translational modifications for efficient targeting. Useful protein substrate tags include, but are not limited to, -202-201207108 IGF2R target (IGF2 (GILT)/IGF2 engineered), transferrin receptor target (transferrin, TfR-target), and Tat protein ( The cell surface is hydrolyzed by heparin heparin proteoglycan (HSPG) mediator Tat). Other proteins that can be used as labels for lysosomal targeting include, but are not limited to, vitamin D binding protein, folate binding protein, lactoferrin, sex hormone binding globulin, transthyretin, sphingolipid activation. Proprotein, retinol binding protein, apolipoprotein B, apolipoprotein E, prolactin, receptor-associated protein (without HNEL sequence in one embodiment), natural transferrin and mutation transfer (eg K225E/ R651A mutation or K225E/K55 3A mutation). In one embodiment, the presentation construct also encodes one or more report sequences, a location tag sequence, and a detection tag sequence. It will further be appreciated that the compositions of the invention or methods of using the compositions can be combined with any chemotherapeutic agent that eliminates, reduces, inhibits or controls the growth of tumor cells or tumors in vivo (eg, to provide a combined therapeutic regimen). . The term "chemotherapeutic agent" as used herein refers to any therapeutic compound that is administered to treat or prevent tumor growth in vivo. In particular, chemotherapeutic agents compatible with the present invention comprise "conventional" chemotherapeutic agents such as small molecules and recently developed biological agents such as antibodies, interleukins, antisense molecules, etc., which are used to reduce or delay The growth of malignant cells. In one aspect, the invention provides a pharmaceutical composition suitable for administration to a human or a non-human, the pharmaceutical composition comprising in vitro engineered to exhibit an immunomodulatory agent such as TNF-[alpha] and/or IL- An immunocyte or TSC population of a functional protein of 12, such as an adenoviral vector, wherein the modulator-203-201207108 is suitable for intratumoral administration. In another embodiment, the composition, e.g., the pharmaceutical composition, comprises a carrier that conditionally exhibits an immunomodulatory agent such as TNF-ct. In some embodiments, the composition comprises a gene transfer vector of about 1 x 1 〇 5 or more particle units (pu). A "particle unit" is a single carrier particle. In certain embodiments, the composition comprises a gene transfer vector of about 1 x 1 〇 6 particle units (eg, about 1×1 〇7 or more particle units, about 1×1 〇8 or more particle units, or about 1×1 09 or more). Multi-particle unit). In other embodiments, the composition comprises about 1x1 〇 1 () or more pu, 1 x 10 11 or more pu, lxlO 12 or more pu, lxlO13 or more pu, lxlO14 or more pu, or lxl 015 Or more pu gene transfer vectors, particularly viral vectors, such as replication defective adenoviral vectors. The number of particle units of the gene transfer vector contained in the composition can be determined by any known suitable method, such as a solution for comparing the absorbance of the composition with a standard solution of the gene transfer vector (ie, a known concentration of the gene transfer carrier) as further described herein. The absorbance of ). The invention further provides a pharmaceutical composition comprising an activating ligand such as RG-1 15819, RG-115830 or RG-115932, wherein the composition is suitable for intraperitoneal, oral or subcutaneous administration. Compositions of the invention, e.g., comprising engineered DC, a carrier (e.g., an adenoviral vector) or an activating ligand, may additionally comprise a pharmaceutically acceptable carrier. The carrier can be any carrier suitable for the engineered dendritic cells, gene transfer vectors or activating ligands. Suitable carriers for the compositions are described in U.S. Patent No. 6,225,289. The carrier is typically a liquid, but can also be a solid, or a combination of a liquid and a solid component. The carrier is deliberately pharmaceutically acceptable (e.g., physiologically or pharmaceutically acceptable) carrier (-204-201207108 such as excipient or diluent). The pharmaceutically acceptable carrier system is well known and readily use. The choice of carrier will be (at least in part) determined by the particular ingredients in the composition and the particular method used to administer the composition. The composition may additionally comprise any other suitable ingredient', particularly to enhance the stability of the composition and/or its end use. Thus, the compositions of the present invention have a wide variety of suitable modulators. Formulations suitable for oral administration include (a) a liquid solution such as an effective amount of the active ingredient dissolved in a diluent such as water, saline or orange juice, (b) a capsule 'pouch or lozenge, each containing a predetermined amount of the active ingredient in solid or granule form, (c) a suspension in a suitable liquid, and (d) a suitable emulsion. Tablets may include one or more of lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, gum arabic, gelatin, colloidal cerium oxide, croscarmellose sodium, talc, magnesium stearate, Stearic acid and other excipients, pigments, diluents, buffers, wetting agents, preservatives, odorants, and pharmaceutically compatible excipients. Ingredients in the form of ingots may contain the active ingredients of the taste, usually sucrose and gum arabic or xanthine, and lozenges containing the active ingredient in an inert base such as gelatin and glycerin or sucrose and gum arabic, and in addition to the active ingredient Emulsions, gums and such analogs comprising excipients known in the art are included. Modulators suitable for administration by inhalation include aerosol formulations. The aerosol formulation can be placed in a pressurized acceptable propellant such as dichlorodifluoromethane, propane, nitrogen and the like. They can also be formulated into unpressurized formulations for administration from a nebulizer or nebulizer. Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic-205-201207108 bacterial injection solutions, which may contain antioxidants, buffers, bacteriostatic agents, and blood for the preparation and target recipients. Isotonic solutes, and may include suspensions, solubilizers, thickeners, stabilizers, and preservatives in aqueous and non-aqueous sterile suspensions. The modulator may be present in unit or multi-dose sealed containers, such as ampoules and vials, and may be stored in a freeze-dried (lyophilized) state that requires only the addition of a sterile liquid excipient such as water for injection prior to immediate use. . Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and the types of lozenges previously described. A preparation suitable for anal administration can be prepared as a suppository by mixing the active ingredient with various bases such as an emulsifying base or a water-soluble base. Formulations suitable for vaginal administration may be presented as pessaries, slivers, creams, gels, pastes, foams or spray formulations which, in addition to the active ingredient, contain carriers known in the art. Additionally, the composition may comprise additional therapeutic or bioactive agents. For example, a therapeutic factor for treating a particular indication can be included. Inflammatory factors such as ibuprofen or steroids may be part of this composition to reduce swelling and inflammation associated with administration of the gene transfer vector in vivo and physiological urgency. The immune system inhibitor can be administered with the composition method to reduce any immune response to the gene transfer vector itself or to the disease. Alternatively, an immunopotentiator can be included in the composition to upregulate the body's natural defense against disease. Alternatively, the interleukin can be administered with the composition to attract immune effector cells to the tumor site. In a particular embodiment described herein, the invention provides a method of treating a tumor, the method comprising the steps in the following order: -206- 201207108 a.  The in vitro engineered immune cells or TSC population are administered intratumorly to the mammal; and b.  A therapeutically effective amount of an activating ligand is administered to the mammal. In one embodiment, the activation ligand system is administered at substantially the same time as a composition comprising an in vitro engineered immune cell or a TSC or a vector, such as an adenoviral vector, for example, by administering the cell or vector. Within 1 hour before or after the object. In another embodiment, the activation ligand system is administered within about 24 hours or less after administration of the in vitro engineered immune cells or TSCs or carriers. In yet another embodiment, the activation ligand system is administered within about 48 hours or less after administration of the in vitro engineered immune cells or TSCs or carriers. In another embodiment, the system is RG-1 1 5932. In another embodiment, the formulation is administered at a dose of from about 1 to 50 mg/kg/day. In another embodiment, the system is administered at a dose of about 30 mg/kg/day. In another embodiment, the formulation is administered daily for a total of 7 to 28 days. In another embodiment, the formulation was administered daily for a total of 14 days. In another embodiment, about lxl 06 to lxl 08 cell lines are administered. In another embodiment, an approximately lxlO7 cell line is administered. In one embodiment, the dendritic cell line is engineered to conditionally express IL-2 and IL-12. IL-2 exhibits a strong immunomodulatory effect on both effector and regulatory T, NK and NK-T cells. It is expected that the expression of IL-2 and IL-12 in cells will result in up-regulation of the receptors with each other and induce different but complementary biological effects by separate signaling pathways. It is also expected that the combination of IL-2 and IL-12 will prolong the duration of immunostimulation and reduce the effective dose of cells to increase animal tolerance -207 - 201207108. See Dietrich 2002, Wigginton 2002, 200 1,1 996 and Koyama, 1 997, McDermott and Atkins 2008; Berntsen et al 1 2 0 0 8; Tarhini et al 1 2008; Heemskerk et al 2008; Horton et al 2008. The polynucleotide sequence of IL-2 can be obtained from accession number U25676 (human), NM_00 8 3 66 (mouse) 'NM_204153 (chicken) and NM_053836 (rat). The polynucleotide sequence of IL-12 can be obtained from accession number NM_000882 (human IL12A), NM - 002 1 87 (human IL12B), NM_0083 5 1 (mouse IL12a), NM_0083 52 (mouse IL12b), NM - 2 1 3 5 8 8 (chicken IL12A), NM — 2 1 3 5 7 1 (chicken IL12B), NM_05 3 3 90 (rat IL12a) and NM — 0226 1 1 (rat IL 1 2b). SEQ ID NO: 13, 15, 21 and 23 encode IL-12 and its subunits in humans and mice. In another embodiment, the dendritic cell line is engineered to conditionally express IL-18 and IL-12. IL-18 induces IFN-γ production and promotes T helper cell development and NK activation. In addition, IL-18 can amplify the production of GM-CSF and reduce the production of IL-10. It is expected that the expression of IL-18 and IL-12 will overcome the limitations observed when administering individual interleukins alone. It is expected that simultaneous expression of IL-12 and IL-18 in dendritic cells will stimulate a stronger tumor antigen-specific Th1 response than when dendritic cells are only transduced by individual interleukins alone. The highest amount of INF-γ production and complete tumor rejection (Tatsumi 2003), which was engineered to simultaneously secrete IL-12 and IL-18, was injected intratumorally. See Vujanovic, 2 0 0 6. See also Coughlin, 1 998, Subleski, 206, Tatsumi, 2 0 0 3, and Sabel, 2007; Shiratori et al 20 07; Lian et al 2007; Iinuma et al 2 006. The polynucleotide sequence of IL-12 is shown above. The polynucleotide sequence of IL-1 8 is available from Accession No. U90434 (human), -208-201207108 NM_0 0 83 60 (mouse), EU74 73 3 3 (chicken), and AY25 8448 (rat). In another embodiment, the dendritic cell line is engineered to conditionally express IL-15 and IL-12. IL-15 has some of the same biological activities as IL-2, thus making IL_15 possible for the treatment of cancer. IL-15 stimulates the proliferation of NK cells and activated T cells and supports the expansion of effector T cells. The report states that IL-15 synergizes with IL-12 to enhance NK cell production of IFN-γ. Koka, 2004; Basak 2008; Lasek et al 2004. In the melanoma model, intratumoral administration of IL-15 and IL-12 induced significant tumor regression (Lasek 1 999). The polynucleotide sequence of IL-12 is shown above. SEQ ID NOS: 11 and 19 encode human and mouse IL-15. Figures 2 and 4 are plastid gene maps that can be used to express the expression systems of human and mouse IL-12 and IL-15. In another embodiment, the dendritic cell line is engineered to conditionally express IL-21 and IL-12. IL-21 and its receptor share sequence homology with IL-2 and IL-15. IL-2 1 promotes the expansion and maturation of NK cells. The biological effects of IL-21 may be synergistic with IL-12, as treatment of NK cells with IL-21 results in a significant upregulation of IL-12 receptor. In addition, IL-21 enhances IL-12 signaling and cooperation to increase IFN-γ production. The polynucleotide sequence of IL-1 2 is shown above. The polynucleotide sequence of IL-21 is available from accession numbers AF254069 (human), NM-021 7 82 (mouse), NM_001024835 (chicken), and NM_001 108943 (rat). SEQ ID NOs: 6, 7, 8, 9 and 17 encode IL-21 in humans and mice. SEQ ID NOS: 1 and 2 are polynucleotide constructs encoding mouse and human IL-12 and IL-21. Figures 7 and 8 are plastid gene maps that can be used to express the expression systems of human and mouse IL-12 and IL-21, respectively. In another embodiment, the dendritic cell line is engineered to present TNF-α and IL-12 in a conditional table of -209 to 201207108. An effective activator of TNF-α immunocytes, whose mediator has anti-tumor properties. In addition, TNF-α can act synergistically with IL-12 to enhance the expression of IFN-γ and IL-12 receptors on T cells. In animal experiments, simultaneous use of IL-12 and TNF-a resulted in tumor infiltration of DN8+ T cells, significant IFN-γ production, and subsequent tumor regression. See Sabel, 2003, 2004, 2007, Taniguchi, 1998, Lasek, 2000; and Xia et al 2008. The polynucleotide sequence of IL-12 is shown above. The polynucleotide sequence encoding TNF-α is available from Accession Nos. X〇2910 (human), NM_0 1 3 693 (mouse), and BC107671 (rat). In another embodiment, dendritic cell lines are engineered. Conditionally expresses IL-7 and IL-12. Members of the IL-7 family of IL-2 are critical for lymphocyte production by T cells and B cells. IL-7 regulates the persistence of the survival and proliferation of the primary and memory CD 8+ T cells. IL-7 has been shown to enhance CTL production against tumors. In addition, IL-12 acts directly on CD8+ T cells to promote proliferation of IL-7 mediators. In addition, it has been reported that IL-7 and IL-12 synergistically enhance the cytotoxicity of CD8+ T cells. Mehrotra, 1 995; Sharma et al 2003; Tirapu et al 2002. Therefore, it is expected that the combined performance of IL-7 and IL-12 will provide a more ideal anti-tumor response. The polynucleotide sequence encoding IL-12 is shown above. The polynucleotide sequence encoding IL-7 is available from accession numbers J04156 (human), NM-008371 (mouse), NM_001037833 (chicken), and NM-0 1 3 1 1 0 (rat). In another embodiment, the dendritic cell line is engineered to conditionally express G Μ - C S F and IL -1 2 . G Μ - C S F regulates the differentiation and proliferation of hematopoietic progenitor cells, and professional antigen-expressing cells (APCs) such as dendritic cells mature on -210- 201207108 play a particularly important role. GM-CSF also enhances the ability of dendritic cells to process and express antigens. GM-CSF differs from IL-12 in function, but both induce significant anti-tumor responses in animal experiments. The combination of IL-12 (T cell activation) and GM_CSF (dendritic cell activation) is expected to result in more potent anti-tumor immunity. In animal studies, the combination of GM-CSF and IL-12 treatment significantly inhibited tumor growth in a variety of cancer models. Wang, 200 1; Chang, 2007; Jean, 2004; Nair, 2006; Hill 2002; Small et al 2007.  In human trials, GM-CSF + IL-12 was successfully used to treat myeloma, but the combination of the two interleukins resulted in a decrease in circulating B cells. Rasmussen, 2003; Hansson, 2007; Abdalla, 2007. It is therefore expected that the co-expression of GM-CSF and IL-12 in a single cell will avoid undesired systemic responses such as circulating B cell reduction. The polynucleotide sequence encoding IL-1 2 is shown above. The polynucleotide sequence of GM-CSF can be obtained from accession numbers M1 1 734 (human), NM_009969 (mouse), EU5203 03 (chicken), Μ Μ _ 0 0 1 0 3 7 6 6 0 (rat Csf2ra) and NM_1 3 3 5 55 (rat Csf2rb). In another embodiment, the dendritic cell line is engineered to conditionally express a chemokine (eg, CCL3 (MIP-la), CCL5 (RANTES), CCL7 (MCP3), XCL1 (lymphocyte chemotactic factor), CCL19 (MIP-3b), CXCL9 (MIG), CXCL 1 0 (IP -1 0), CXCL 1 2 (SDF-1) or CCL2 1 (6Ckine)) and IL-12. Chemokines are chemotactic interleukins that regulate the movement and activation of white blood cells and other cell types under a variety of inflammatory and non-inflammatory conditions. Inflammatory interleukins control the recruitment of white blood cells in inflammation and tissue damage. Constitutive chemokines function as housekeepers, such as guiding the entry and retention of white blood cells (such as dendritic cells) into secondary lymphoid organs and the bone marrow and thymus during the blood period of 211 - 201207108. In animal studies, intratumoral injection of two adenoviruses that separately express IL-12 and CXCL10 resulted in regression of tumor nodules derived from the CT26 murine colorectal cancer cell line. Narvaiza et al., 2000. Emtage et al. , 1999 describes two dual recombinant adenoviral vectors that express IL2 and XCL1 (lymphocyte chemotactic factors) or IL-12 and XCL1. In breast adenocarcinoma tumors of mice, intratumoral injection of the vector induces a potent anti-tumor response and leads to a protective immune response. In other animal experiments, co-transduction of adenoviral vectors expressing IL-12 and CCL27 resulted in tumor regression and long-term specific immune responses. Gao et al., 2007. Therefore, it is expected that the common expression of chemokines and IL-12 in the present invention will result in synergistic antitumor activity. In another embodiment, the dendritic cell line is engineered to conditionally express anti-angiogenic interleukins (e.g., IP-10 and Mig) and IL-12. IP-10 and Mig are chemotactic agents for T cells and NK cells, and their ability to inhibit angiogenesis depends on NK cells. Animal experiments have shown that combination therapy with two adenoviruses, one of which shows IP 1 〇 'another expression of IL -1 2 , results in significant anti-tumor co-saturation. Narvaiza et al. , 2000. In other assays, adenoviral vectors expressing ιριο or MIG and/or IL-1 2 were administered intratumorally into a murine model of breast cancer and fibrosarcoma. The trial found that combined administration of IP or IG and IL-1 in combination with IP10, MIG, IL-1 2 alone or in control treatment resulted in significant tumor regression and increased survival time of tumor-bearing animals. Among them, the combination of 1?-1〇 and 11^12 is most effective. ? 31!^1:, 2001. See also Mazzolini, 2003 and Huang 2004. It is therefore expected that a common expression of anti-angiogenic interleukins and IL-1 2 will result in synergistic anti-tumor activity. To demonstrate potent IL-12-media gene therapy, the conditional -212 - 201207108 cDNA expression system was used to allow for the opening of immune cells or TSC-producing immunomodulators and/or IL- at different time points after intratumoral injection. 12. Based on the results in the invasive B16 melanoma model of C57BL/6 mice, the following conclusions were reached: 1) The amount of IL-12 increase was determined by DC. RheoIL12 is secreted in the presence of the activating ligand RG- 1 1 5 83 0, but is not secreted when the ligand is absent; 2) intratumoral DC. RheoIL12-basal therapy with intratumoral DC. CIL12-basal therapy is as effective as RG-1 1 5 83 0 is administered to treated animals within 24 hours of DC injection (providing ligands at later time points, failure of RG-1 1 5 83 0 therapy) 3) The performance of IL-12 in DC appears to prolong the survival of these cells in the tumor microenvironment and is associated with a greater number of intratumoral injections of DCs that move to the lymph nodes of tumor drainage; and 4) related to treatment outcomes; The strongest immune response is the amount of tumor-specific CD 8+ T cells treated for cross-sensitization rather than the amount of injected DC left in the tumor microenvironment. Overall, these materials show DC. ILI2-basal therapy may be successful, based on their positive effects on input (cross-sensitization) type 1 CD8+ T effector cells rather than subsequent export events, such as injection of DC-mediated recruitment of anti-tumor T cells to the tumor microenvironment. These cells (immune cells or TSCs) may be treated with a factor to stimulate the activity of the cells prior to intratumoral injection. For example, such cells may be treated with costimulatory molecules, such as positive costimulatory molecules including OX40L, 4-1BBL, CD40, CD40L, GITRL, CD70, LIGHT or ICOS-L or negative co-stimulatory molecules such as anti- CTLA4, anti-PD-L1 or anti-PD-L2 antibody. For example, such cells (e.g., immune cells or TSCs) may be cultured with cells expressing one or more costimulatory molecules, e.g., -213-201207108 J5 88 lymphoma cells that exhibit CD40 ligand molecules. In another embodiment, the cells (immune cells or TSCs) may be treated with a reverse immunosuppressive molecule (tolerance inhibitor) such as an anti-TGF-beta antibody (for inhibition of TGF in the microenvironment) Co-), anti-IL10 antibody, TGFbRII DN (to inhibit TGF signaling in gene-modified cells), IL-10R DN, dnFADD (to inhibit intracellular cell death pathway), anti-SOCS1 antibody, siRNA or bait ( To inhibit intracellular inhibitory interleukin signaling or anti-TGFa antibodies. The recombinant adenovirus carrying the polynucleotide sequence shown in Figures 1 to 8 is produced. For example, hIL-2 1 is a co-transfected hIL-2 1 expression vector that linearizes the site upstream of the left ITR by restriction enzymes and an appropriate (eg, E3 deletion) adenoviral backbone to the permissive cell line. Prepare such as HEK293 cells. An adenoviral vector carrying a murine immunomodulator gene was used to transduce murine dendritic cells or TSC for use in a murine therapeutic model. In human therapeutic applications, a polynucleotide encoding a human homologous immunomodulator gene is inserted into a suitable vector. The adenoviral vector for therapeutic use in humans is produced under conditions of GMP. Examples of treatment outlines (clinical trials) for stage III/IV melanoma patients are as follows: The treatment of this case involves intratumoral injection of adenovirus-transduced dendritic cells and oral administration of activated drugs for 14 days (with body). Subjects were screened for 30 days until one week prior to the start of the clinical trial. Each subject is required to sign a subject consent before any processing procedures are performed. The program host will inform all subjects about the nature, purpose, timing, potential hazards, and procedures to be performed during the trial, and the subject's medical records may be subject to F D A review. Subjects (total 16 to 20) were randomly assigned -214-201207108 to 4 groups. All groups received intratumoral injections of up to 5 x 107 transduced dendritic cells approximately 3 hours after the first oral administration of the ligand. The daily oral doses of the four groups of ligands administered were different: for example, group 1 = 〇.  〇 1 mg / kg, group 2 = 0. 3 mg/kg, group 3 = 1 mg/kg, group 4 = 3 mg/kg. During treatment, blood is drawn at specific times to assess single dose and steady state pharmacokinetics of the activating drug and its major metabolites. Similarly, blood was drawn at specific time points to assess the humoral and cellular immune responses to the viral vector, RTS components, and tumors. Urine and blood were collected at specific time points for serum chemistry 'urine analysis and hematology (safety check). Tumor and/or draining lymph node biopsy is performed at specific time points to assess transgene performance and immune response to the tumor as a result of treatment. The condition for early termination of the test is an adverse reaction to the patient and the adverse reaction is recorded. Patients were followed for 1, 2, 3, and 4 months to observe adverse events and treatment outcomes. In another embodiment, a system in need of treatment of a tumor (a) is administered to a dendritic cell engineered to constitutively or conditionally express an immunomodulatory agent, such as an immunomodulatory agent disclosed herein, and/or ( b) A constitutive or conditionally indicative immunomodulatory agent, such as an immunomodulatory agent disclosed herein, is injected intratumorally into the subject. In one embodiment, the dendritic cell line is engineered to represent an Ad-immunity modulator vector, particularly an Ad-RTS-immunity modulator vector. In another embodiment, the vector is injected intratumorally into an individual, an Ad-immunomodulator vector, particularly an Ad-RTS•immunomodulator vector. In another embodiment, a system for treating a tumor is required (a) to be subjected to 215-201207108 for constitutive or conditional expression of IL-1 2 dendritic cells, and (b) constitutive or The vector for conditionally expressing IL-12 is injected intratumorally into the individual. In one embodiment, the dendritic cell line is engineered to express an Ad-IL-12 vector, particularly an Ad-RTS-IL-12 vector. In another embodiment, the vector that is injected intratumorally into the individual is an Ad-IL-12 vector, particularly an Ad-RTS-IL-12 vector. In another embodiment, a system in which a tumor is to be treated (a) is administered to a dendritic cell engineered to constitutively or conditionally express IL-1 2, and (b) the system is administered or A plurality of anti-cancer chemotherapeutic agents. In one embodiment, the engineered dendritic cell line is engineered to express an Ad-IL-1 2 vector, particularly an Ad-RTS-IL-12 vector. A plurality of anti-cancer chemotherapeutic agents can be administered prior to administration of the engineered dendritic cells, after administration of the engineered dendritic cells, or while administration of the engineered dendritic cells. In another embodiment, the anticancer chemotherapeutic agent is pac lit axel, paclitaxel derivative or analog, temozolomide (tem ο ζ ο 1 omide), temozolomide derivative or analog, sulphonic acid Sunitinib, sunitinib derivative or analog, gemcitabine, or gemcitabine derivative or analog. In another embodiment, a system for treating a tumor is required (a) by administration Engineering to express IL-1 2 constitutively or conditionally Dendritic cells' and (b) constitutively or conditionally expressing IL-12 are injected intratumorally into the individual, and (c) the system is administered with one or more anti-cancer chemotherapeutic agents. In the aspect, the dendritic cell line is engineered to express the A d -1 L - 1 2 vector, particularly the Ad-RTS-IL-12 vector. In another embodiment, the tumor is treated with -216-201207108 The vector for injection into an individual is an Ad-IL-12 vector, particularly an Ad-RTS-IL-12 vector. The one or more anti-cancer chemotherapeutic agents can be administered to the engineered dendritic cells and the expression IL- Before the carrier of 12, after administration of the engineered dendritic cells and the vector expressing IL-1 2, or while administering the engineered dendritic cells and the vector expressing IL-12 In one embodiment, the anticancer chemotherapeutic agent is paclitaxel, paclitaxel derivative or analog, temozolomide, temozolomide derivative or analog, sunitinib , sunitinib derivative or analogue, gemcitabine, or gemcitabine In another embodiment, a system in which a tumor is to be treated (a) is administered to a dendritic cell engineered to constitutively or conditionally express an immunomodulatory agent, such as an immunomodulatory agent disclosed herein. And (b) the system is administered with one or more anti-cancer chemotherapeutic agents. In one embodiment, the engineered dendritic cell line is engineered to represent an Ad-immunity modulator vector, particularly Ad -RTS-immunomodulatory agent. The one or more anti-cancer chemotherapeutic agents can be administered prior to administration of the engineered dendritic cells, after administration of the engineered dendritic cells, or upon administration of the Engineered dendritic cells are administered simultaneously. In one embodiment, the anticancer chemotherapeutic agent is paclitaxel, paclitaxel derivative or analog, temozolomide, temozolomide derivative or analog, sunitinib, sulphonic A derivative or analog of analogy, gemcitabine, or a gemcitabine derivative or analog. In another embodiment, a system for treating a tumor is required (a) to be subjected to a dendritic cell engineered to constitutively or conditionally exhibit an immunomodulatory agent such as the immunomodulatory agent disclosed herein, (b) constitutively or conditionally expressing an immunomodulatory agent, such as an immunomodulatory agent disclosed herein, is injected intratumorally into the subject, and (c) the system is administered with one or more anti-cancer chemotherapeutic agents. In one embodiment, the dendritic cell line is engineered to represent an Ad-immunity modulator vector, particularly an Ad-RTS-immunomodulator vector. In another embodiment, the vector that is injected intratumorally into the individual is an Ad-immunity modulator vector, particularly an Ad-RTS-immunomodulator vector. The one or more anti-cancer chemotherapeutic agents can be administered to the engineered dendritic cells and the vector exhibiting the immunomodulatory agent prior to administration of the engineered dendritic cells and the vector exhibiting the immunomodulatory agent Thereafter, or after administration of the engineered dendritic cells and the vector exhibiting the immunomodulatory agent. In one embodiment, the anticancer chemotherapeutic agent is paclitaxel (pac 1 ita X e 1), paclitaxel derivative or analog, temozolomide, temozolomide derivative or analog, sulphonic acid Sunitinib, sunitinib derivative or analog, gemcitabine, or gemcitabine derivative or analog. In any of the methods of the invention, the disease or condition can be the disease or condition disclosed in this application. In one embodiment, the disease or condition is listed in the disease or condition of Table 1 herein. In another embodiment, the disease or condition series is shown in the disease or condition of Table 3 herein. In any of the methods of the invention, the cancer or tumor can be a disease or condition disclosed in the present application. In one embodiment, the cancer or tumor series is shown in the cancer or tumor of Table 1 herein. In another embodiment, the cancer or tumor - 218 - 201207108 tumor series is shown in the cancer or tumor of Table 3 herein. It is possible to measure the effect of the immunomodulatory agent and/or IL-1 2 on the cell population by measuring the amount of expression or activity of the Th1/Tcl-type interleukin IFN-γ in the biological sample from the patient. For the purposes of the present invention, the present invention provides a method of determining the therapeutic effect of a therapeutic formulation based on a living body engineered immune cell or TSC on a cancer patient, the method comprising: a. Measuring the amount or amount of interferon gamma (ΙΡΝ-γ) in the first biological sample or the amount of both, the first biological sample being administered to the in vitro engineered cell such as immune cells or TSC Previously obtained from a human patient, thereby generating a control amount; b. administering the in vitro engineered cells to the patient via the tumor; c. administering an effective amount of the activating ligand to the patient; d.  Measuring the amount or amount of IFN-γ or the amount of both in the second biological sample. The second biological sample is obtained from the patient after administration of the activating ligand, thereby generating a detection amount; e.  Comparing the control amount and the detection amount of IFN-γ, wherein the amount of the expression of the IFN-γ, the amount of the activity, or the amount of the two detected is increased compared to the control amount, indicating that the therapeutic treatment formula is effective for the patient line. . The invention may also optionally include the following additional steps: f.  Obtain a biopsy sample and calculate tumor infiltrating lymphocytes (TIL), and/or g.  Observe the tumor regression in response to treatment. The term "individual" means a complete insect, plant or animal. When the system is -219-201207108 fungal or yeast, it is also expected that the ligand will function equally. Animals which can be used in the present invention include, but are not limited to, vertebrates such as mammals such as humans, rodent 'monkeys and other animals, of which human or mouse are preferred. Other animals include veterinary animals such as dogs, cats, horses, cows, sheep, goats, pigs, and the like. The present invention further provides a method of increasing the expression of an immunomodulatory agent, such as TNF-[alpha], by introducing one or more regulatory sequences and a nucleic acid which can optionally encode a signal peptide to a vector, such as a replication defective adenoviral vector, wherein the vector The immunomodulator is conditionally expressed. The term "protein expression" as used herein includes, but is not limited to, transcription, post-transcription, translation, and/or post-translation. The invention also encompasses a method of increasing the expression of an mRNA or protein of an immunomodulatory agent, such as TNF-[alpha], which method comprises producing a vector which conditionally expresses TNF-a, wherein the vector further comprises one or more encoding the TNF-a The polynucleotide sequence is ligated to a regulatory sequence, and an activating ligand is added, thereby inducing expression of the immunomodulatory agent, wherein the one or more regulatory sequences and/or signal peptides enhance the expression of the TNF-a. Various regulatory regions of the invention include, but are not limited to, a 5' non-translated region (5' UTR), a 3' UTR, or both. In one embodiment, the '5' UTR is a 5U2. 5U2 fusion dog SERCA2 intron 2, which has a poly-Α site with a mutation-consistent consensus, located at the 5' end of the exon 2 splicing donor site and The exon 3 is located at the 3' end of the excision, followed by a portion of the 5' UTR of the casein. In another embodiment, the 3' regulatory region is a polyadenylation signal of SV4 or hGH. In certain embodiments, the methods of the present invention are also directed to enhancing the secretion of TNF-a by the production of a conditional TN F - a vector, wherein the vector -220-201207108 further comprises a signal peptide, thereby The secretion of TNF-[alpha] is increased compared to a vector comprising a TNF-[alpha] natural signal peptide gene, such as a TNF-[alpha] wild type signal peptide. In particular, the signal peptides used in the present invention are codon-optimized. In a particular embodiment, the signal peptide is encoded by an IL-2 wild type signal peptide gene. In another specific embodiment, the signal peptide is encoded by a codon-optimized IL-2 signal peptide gene. While not wishing to be bound by theory, it is contemplated that the present invention will support the use of in vitro engineered immune cells and TSC-based gene therapy for clinical trials, with objective clinical response as the primary end point, and crossover. Sensitized anti-tumor CD8+ T cells (producing IFN-[gamma]) are secondary endpoints. The ability to turn on or off the performance of immunomodulators and/or IL-1 2 in vivo adds an element of safety and therapeutic control to the treatment, since the timing and amount of protein expression can be controlled by the administration of the ligand, and immunization The timing of modulators and/or IL-12 expression is also expected to be critical to the therapeutic effectiveness of the method. The present invention further supports the use of living, extracorporeally engineered cells that conditionally express genes of interest as an innovative means for the therapeutic and therapeutic use of human diseases in a useful and efficient manner. The invention also provides a method of treating a tumor, reducing tumor size, or preventing tumorigenesis in a mammal in need thereof, in which a carrier of a protein having the function of one or more immunomodulators is conditionally represented (but the vector does not comprise In the cells) is administered intratumorally to the tumor microenvironment. In this embodiment, the vector is administered to the tumor in a form that is not packaged in a cell such as an immune cell or TSC. The present invention also provides a method of treating a disease in a mammal in need thereof, in which a protein carrier having a function of one or more immunomodulators is present (but the vector is not contained in the cell) ) is administered to the mammal. In this embodiment, the vector is administered to the tumor in a form that is not packaged in a cell such as an immune cell or TSC. In one embodiment, the immune cells, TSCs, dendritic cells or bone marrow dendritic cells are not administered into the tumor together with the vector. In another embodiment, the vector of the present invention which is not contained in the cells is administered intratumorally, simultaneously with, before or after the immune cells, TSCs, dendritic cells or bone marrow dendritic cells are administered intratumorally. In one embodiment, the vector of the present invention which is not contained in the cells is administered intratumorally to the same lesion as that administered by the immune cells or TSC. In another embodiment, the vector of the present invention which is not contained in the cells is administered intratumorally to a lesion different from that administered by the immune cells or TSC. In one embodiment, the vector is administered to the same lesion each time the administration cycle. In another embodiment, the administered vector is not administered to the same lesion every administration cycle. In one embodiment, the tumor is, for example, any of the tumors listed in Tables 1 and 3. In another embodiment, the tumor is a melanoma, a colorectal tumor, a pancreatic tumor, a breast tumor, a lung tumor, or a kidney tumor. In another embodiment, the tumor is a malignant melanoma. In another embodiment, the tumor is a 111 C or IV stage malignant melanoma. In one embodiment, the dose of the carrier cycle per tumor is at least about 1 · Ox 1 〇 9 virions. In another embodiment, the dose of the carrier cycle administered to the tumor at each time from -222 to 201207108 is at least about ιοχ10&quot; virions. In another embodiment, the dose of the vector cycle administered intratumor is about 1. OxlO9 to about l. OxlO13 virions. In another embodiment, the dose of the vector cycle administered intratumor is about 1 〇χΐ〇 1 () to about 1. 0 X 1 0 13 virions. In another embodiment, the dose of the vector cycle administered intratumor is about i. 〇xi〇1Q, about ι. Οχ10&quot;, about ι·0χ1〇12 or about 1. 0x10 13 virions. In one embodiment, the vector is ad-RTS-IL-12. In another embodiment, the invention further provides a method of treating liver disease in a mammal in need thereof, wherein the carrier for conditionally expressing the protein is not contained in the cell, and the vector is administered to the Mammalian 0 In another embodiment, the invention further provides a method of treating a lysosomal storage disease in a mammal in need thereof, wherein the carrier for conditionally expressing the protein is not contained in the cell, and The vector is administered to the mammal. In another embodiment, the invention further provides a method of treating a disease in a non-human mammal in need thereof, wherein the carrier for conditionally expressing the protein is not contained in the cell, and the vector is administered to The dose of the mammalian oxime activating ligand is about 5 to 100 mg/day, for example about 5, 10, 15 , 20 ' 25 , 30 ' 35 , 40 , 45 , 50 , 55 , 60 ' 65 , 70 , 75 , 80, 85, 90, 95 or 1 mg/day. In one embodiment, the active formulation system is administered at least once a day. In another embodiment, the live-223-201207108 compounding system is administered once daily for about 14 days. In one embodiment, at least two carrier doses (eg, about 1×1 〇11 and 1×10 1 2) are at least three different activating ligand doses (eg, from about 5 mg/day to about 100 mg/day). Used in combination. One of ordinary skill in the art will be able to optimize the dosage to provide a range of effective carrier plasma levels to the extent that various activating ligands are activated. In one embodiment, the dosage of the activating ligand administered to the individual varies with the time of administration of the activating ligand within the intratumoral delivery cycle. In another embodiment, the dosage of the activating ligand administered to the individual is reduced with the time of administration of the activating ligand within the intratumoral vehicle administration cycle. In another embodiment, the dosage of the activating ligand administered to the individual is increased (gradually increased) with the time of administration of the activating ligand within the intratumoral administration cycle. In one embodiment, the system is treated with a 2, 3, 4, 5, 6, 7, 8, 9 or 1 intratumoral carrier administration cycle. In another embodiment, the system is treated with 3 to 7 intratumoral vehicle administration cycles. In another embodiment, the system is treated with 4 to 6 intratumoral vehicle administration cycles. In another embodiment, the system is treated with 5 or 6 intratumoral vehicle administration cycles. In another embodiment, the system is treated with 6 intratumoral vehicle administration cycles. In one embodiment, each intratumoral carrier administration cycle is separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In another embodiment, the intra-tumor vector administration cycle is separated by 4 weeks. In one embodiment, the vector dose is changed from -224 to 201207108 for each subsequent intratumoral vector administration cycle. In another embodiment, the carrier dose for each subsequent intratumoral carrier administration cycle is reduced. In another embodiment, the vector dose of each subsequent intratumoral vector administration cycle is increased. In one embodiment, the dose of the carrier and the activating ligand, the number and length of the intratumoral carrier administration cycle, the frequency of carrier administration, and the frequency of administration of the activating ligand are set forth in Table 8 in Example 11. In one embodiment, the invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a carrier of the invention that is not included in the cell. Suitable carriers include, but are not limited to, saline, distilled water, sodium chloride solution, mixtures of sodium chloride and inorganic salts or similar mixtures thereof, solutions of substance solutions such as mannitol, lactose, dextran and glucose, amino acids Solutions such as glycine and arginine solutions, organic acid solutions or mixtures of salt solutions and dextrose solutions, aqueous and non-aqueous isotonic sterile injectable solutions, which may contain antioxidants, chelating agents, buffers, bacteriostats And the preparation is isotonic solute, and may include suspensions, solubilizers, thickeners, stabilizers and preservatives in aqueous and non-aqueous sterile suspensions. The modulator may be present in unit or multi-dose sealed containers, such as ampoules and vials, and may be stored in a freeze-dried (lyophilized) state which requires only the addition of a sterile liquid carrier such as water for injection prior to immediate use. In the event of any conflict between any of the references cited herein and any disclosure or suggestion of this specification, for the purposes of the present invention, all of the patents, patent applications and disclosures cited herein are hereby incorporated by reference. The reference method is fully integrated here. The present invention is not intended to limit the scope of the present invention in any way, and the scope of the claims herein is intended to cover all embodiments and example. U.S. Patent Application Serial No. 1 2/247,73, filed on Oct. 8, 2008, entitled &lt;RTI ID=0.0&gt;&gt; U.S. Patent Application No. 12/24 1,0 18, entitled "Therapeutic Gene Switch Constructs and Bioreactors for Biotherapeutic Molecular Expressions and Their Uses", also filed in this reference. [Embodiment] Example 1 An experiment was conducted to determine the dose and the most effective interleukin of a dendritic cell capable of inducing a tumor-specific immune response and antitumor activity in a Renca renal cell carcinoma tumor model. It was used in this assay: Renca and Renca-HA. The latter cell line was prepared by transfecting Renca cells with influenza virus hemagglutinin (HA). The advantage of the Renca-HA model is the ability to track antigen-specific T cells. Because both CD8 and CD4-specific HA-derived epitopes are known and have been used.Specific Purpose - To determine the HA-specific immune response induced by intratumoral administration of dendritic cells.

The Renca-HA tumor line was established subcutaneously in BALB/c mice. When the tumor grows to be sensible, dendritic cells are injected intratumorally. Dendritic cell administration was repeated twice in 7-226-201207108 days for a total of 3 doses. The following mice were grouped (each group included 3 mice): 1 · untreated mice; 2- treated mice with 5xl05 of control plastid-transduced dendritic cells; 3. with 1 〇6 via control plastids Transduced dendritic cells were used to treat mice; 4- mice were treated with 5xl06 of plastid-transduced dendritic cells; 5. As in groups 2 to 4, IL-12-transduced dendritic cells were used; As in groups 2 to 4, IL-15 transduced dendritic cells were used; and 7. As in groups 2 to 4, IL-21 transduced dendritic cells were used. To test the effect of different combinations of interleukins, mice received the following treatments: 8. 5xl05 dendritic cells transduced with IL-12 and dendritic cells transduced with IL-15 by 5xl05, 9. 5xl05 by IL- 12-transduced dendritic cells and 5xl05 IL-21-transduced dendritic cells, and 10.5xl05 IL-15-transduced dendritic cells and 5xl05 IL-21-transduced dendritic cells. Four days after the last administration, the lymph nodes of the tumor-bearing mice were collected and stimulated with MHC type I conforming peptide (to detect CD8+ T cell response) or MHC type I conformative peptide (to detect CD4+ T cell response). The following assays were used: 1. ELISPOT IFN-γ and IL-2; 2. T-cell proliferation; 3. Detection of TNFa, IL-10, IL-4 and GM--227-201207108 CSF released by lymph node cells. In addition, NK activity of lymph node cells was evaluated using YAC cells as a target. At the same time, cells were stimulated with anti-CD3/CD28 antibody to assess the non-specific response of T cells. The most effective dendritic cell dose that induces an antigen-specific immune response is determined. Specific Purpose 2 - To evaluate the antitumor activity of dendritic cells transduced by interleukin genes. Only those interleukin-transduced dendritic cells that were shown to induce a statistically significant immune response were used for further experiments.

The treatment of Renca-HA tumor-bearing mice was carried out as described in Specific Purpose 1. Interleukin-transduced DCs were used in previous experiments showing doses of specific activity. The control group used dendritic cells transduced with control adenovirus. To achieve statistical significance, each group included 1 mouse. Assess tumor growth. Renca-HA tumors contain immunogenic epitopes that can be used for immunological monitoring and preliminary testing of anti-tumor effects. However, in order to confirm the potential anti-tumor activity of the treatment, untransfected tumor cells must be used. Therefore, the Rene a tumor model was used to repeat the above experiment. Example 2

Intratumoral injection is engineered by adenovirus transduction to demonstrate the safety, tolerability, transgenic function, and immune effects of autologous dendritic cells of hIL-12 and one or more other immunomodulators under the control of RTS. It was evaluated in the melanoma individuals of Dijon and IV-228-201207108 using the methods described below. Subjects in this trial were subjects with stage I and IV melanoma 'subjects will be divided into 4 groups, each subject undergoing a single intratumoral injection (injected to melanoma tumors) engineered by adenovirus transduction The dendritic cells (DC) are dosed with autologous dendritic cells expressing human interleukin 12 (hIL-12) and one or more other immunomodulators (re-injected into the same subject as the cells) 5xl07, along with a daily oral dose of an activating drug (activating ligand). This assay will use an injection of dendritic cells transduced with an adenoviral vector in vitro (after the cells are removed from the subject) to inducibly express human IL-12 and one or more other immunomodulating agents. The function of the injected DC-produced IL-12 and one or more other immunomodulators is "turned on" (induced) by orally administering the activated drug (RG-1 15932) to activate RTS. Assessment of safety and tolerability includes physical examination (including ECOG physical status), vital signs monitoring, serum chemistry, urine analysis, hematology, adverse reactions "side effects" and adenovirus, RTS components and activated drugs Antibodies and cellular immune responses. To assess the progress of the treatment, a single dose of the orally active drug and its major metabolites and steady state pharmacokinetics/ADME, amount of hIL-12 in the target tumor, draining lymph nodes, and peripheral samples of the peripheral circulation, other immunomodulations will be measured. The amount of the agent and the analysis of cellular immune response (T cells) and serum interleukin analysis. For example, 16 patients with stage III and IV melanoma were divided into 4 groups, groups 1 and 2 contained 3 patients, and groups 3 and 4 contained 5 patients. All subjects will receive a single intratumoral injection of 5x107 autologous DC, which is transduced with an adenoviral vector to encode human IL-12 and one or more other exemption -229-201207108 virulence modulators under the control of RTS. For example, a subject receives an intratumoral injection transduced with an adenoviral vector to encode an autologous DC of human IL-12 and an immunomodulatory agent such as IL-15 or IL-21 under the control of RTS. Subjects will receive a single oral dose of active drug per dose (Group 1: 0.01 mg/kg, Group 2: 0.1 mg/kg, Group 3: 1.0 mg/kg or Group 4: 3 mg/kg) The first dose was administered approximately 3 hours prior to DC injection on Day 1, followed by continuous administration for 13 days. Subjects who met the retreatment criteria received additional adenoviral transduced autologous dendritic cell injection in combination with a 14-day daily single oral dose of activating drug therapy. Subjects in Group 1 were assessed for safety, tolerability, and dendritic cell function within 1 month after injection of in vitro engineered dendritic cells. One of the highest doses of activated drugs. The safety assessment will be continued for 3 months after all subjects have injected the engineered dendritic cells for the first time. If toxicity is observed or the subject receives additional dendritic cell injection, the tracking period may be extended to a total of The safety of the subjects was monitored for 6 months. This trial demonstrates the safety and tolerability of single or multiple intratumoral injections of adenovirus-transduced autologous dendritic cells in combination with oral activating drugs for melanoma patients. This test provides a steady state pharmacokinetic/ADME of an orally activated drug. This assay demonstrates the functionality of RTS in an individual by measuring the activation of an adenovirus-transduced autologous dendritic cell in a target tumor and/or draining lymph node in response to oral administration of an activated drug by RT S. L _ 1 2 and the performance of one or more other immunomodulators. In addition, the test confirmed the immunological effects of adenovirus-transduced autologous dendritic cells -230 - 201207108 in the target tumor, draining lymph nodes, and peripheral circulation in terms of cellular immune response after oral administration of the activating drug. Melanoma has been chosen as an exemplary cancer, especially for melanoma. In parenchymal tumors, melanoma specifically shows an effect on immunotherapy, and melanoma tumors are easy to perform intratumoral injection and biopsy. Subjects included in the trial were suffering from unresectable stage III or IV melanoma with a tumor diameter of at least 0.5 cm. The thickness of the tumor was not limited, affecting the number of lymph nodes, showing metastasis or distant metastasis. Preparation of adenovirus containing RheoSwitch therapeutic system, hIL-12 and one or more other immunomodulators Recombinant DN A was transferred to dendritic cells (DC) by transduction of an in vitro adenoviral vector. Recombinant DN A is used to display human IL-12 (p70) and one or more other immunomodulators from immature dendritic cells injected intratumorally to allow DCs to survive and stimulate maturation in the tumor environment and result in subsequent migration of DCs to Drain the lymph nodes. This causes T helper cells to be differentiated into a Th1 type, and at the same time, the activation of tumor-specific cytotoxic T cells is caused by cross-sensitization of tumor antigens.

Recombinant DNA used as a recombinant adenoviral vector allows human IL-12 and one or more other immunomodulators to behave under the control of the RheoSwitch® therapeutic system (RTS). RTS contains bicistronic information from the human ubiquitin C promoter and encodes two fusion proteins: Gal4-EcR and VP16-RXR. The DNA-binding domain of Gal4-EcR yeast Gal4 (amino acid 1 to 147) is fused to the DEF domain of the ecdysone receptor from the insect Chorstoneura fumiferana. In another embodiment, RTS-231 - 201207108 consists of bicistronic information represented by the human ubiquitin C promoter and encodes two fusion proteins: GaU-EcR and VP16-RXR. The DNA-binding domain of Gal4-EcR yeast Gal4 (amino acid 1 to 147) is fused to the DEF domain of the ecdysone receptor from the insect, Choristoneura fumiferana. The VP16-RXR is a transcriptional activation domain of HSV-VP16 fused to the EF domain derived from the chimeric RXR of human and aphid sequences. These Gal4-EcR and VP16-RXR sequences are separated by an internal ribosome entry site (IRES) from EMCV. When Gal4-EcR binds to a small molecule drug (RG-1 1 5 93 2), the two fusion proteins dimerize and self-contained Gal4-reactive with 6 Gal4-binding sites and a synthetic minimal promoter. The promoter activates transcription of hiL-12 with one or more other immunomodulators. The RTS transcription unit described above is placed downstream of hIL-12 and one or more other immunomodulator transcription units. This entire RTS-hIL 12-immunomomodulator cassette was incorporated into the E1 region of the adenovirus 5 genome where it was deleted. The adenoviral backbone also lacks the E3 gene. The genetic map of the adenoviral vector Ad-RTS-hIL-12 is shown in Figure 8 of US 2009/0123441 A1. The recombinant adenoviral vector used in this assay contains, in addition to the viral vector sequence, the following exemplary regulatory elements: the human ubiquitin C promoter, an internal ribosome entry site derived from EMCV, comprising six Gal4-binding sites, Three sets of SP-1 binding sites and an inducible promoter of a synthetic minimal promoter sequence, an SV40 polyadenylation site, and a transcription termination sequence derived from a human alpha-globin gene. It should be understood that other adjustment elements can be used instead. An exemplary recombinant adenoviral vector Ad-RTS-hIL-12-immunomodulator is produced in the following manner. The coding sequence of the receptor fusion proteins VP16-RXR and Gal4-EcR, which are opened by EMCV-IRES (internal ribosome entry site) -232-201207108, was inserted into the adenoviral shuttle vector and received the human ubiquitin C promoter (constitutive Control of the promoter). Next, the p40 and p35 subunits of hIL-12 separated by IRES and the coding sequence of one or more other immunomodulators are inserted into the ubiquitin under the control of a synthetic inducible promoter comprising 6 sets of Gal4-binding sites. Upstream of the C promoter and acceptor sequences. The shuttle vector comprises an adenoviral serotype 5 sequence from the left end to the gene map unit 16 (mul6), after which the E1 sequence is deleted and consists of RTS, IL-12 and one or more other immunomodulator sequences (RTS-hIL-12) ) replaced. The shuttle vector carrying the RTS-hIL-12-immunomodulator was transiently transfected into HT-1 0 80 cells to test the performance of activated drug-dependent IL-12 and other immunomodulators. The shuttle vector was then recombined with the adenoviral backbone by co-transfection into HEK 293 cells to obtain a recombinant adenovirus Ad-RTS-hIL-12-immunomodulator. The adenoviral backbone contains the deletion of the mu 0 to 9.2 on the left side of the genome and the sequence of the E3 gene. The shuttle vector and adenoviral backbone comprise overlapping sequences from mu 9.2 to mu 16, thus allowing for recombination and production of recombinant adenoviral vectors among them. Since the recombinant adenoviral vector lacks the E1 and E3 regions, the virus is defective in replication in normal mammalian cells. However, the virus can replicate in HEK 293 cells containing the adenovirus-5 E1 region thus providing E1 trans-acting. Exemplary recombinant adenoviral vectors are produced by a linearized shuttle vector carrying a DNA element for inducible expression of human IL12 and one or more other immunomodulators and an adenoviral backbone co-transfected into ΗEK293 cells. Recombination between the shuttle vector and the overlapping sequences of the viral backbone results in the production of recombinant adenoviruses and is packaged into viral particles in ΗΕΚ293 cells. ΗΕΚ293 -233- 201207108 The cell line was grown in D Μ EM containing fetal bovine serum. The virus used in the proposed assay was purified by CsCl density gradient centrifugation. The recombinant adenovirus is purified by secondary plaque, and the resulting seed preservation solution is expanded for production of the protovirus pool (MVB) by HEK2 93 cells from a fully characterized stock cell pool. The MVB undergoes extensive cGMP/GLP release testing, including replication-type adenovirus (RCA) 'aseptic, mycoplasma, opportunistic virus, retrovirus, human virus HI VI/2, HTLV1/2, HAV, HBV, HCV , EBV, B19, CMV, HHV-6, 7 and 8, bovine and porcine viruses, intact vector sequencing and the function of IL-12 and one or more other immunomodulators by AD-induced in human cell lines Sex detection. The virus from MVB can be used to produce purified virus in CGMP plants and may be tested again for release, including identification, RCA, sterility, mycoplasma, opportunistic virus, viral particle to infectious unit ratio, host cell Contamination of DN A, endotoxin and protein and functional detection of IL-12 and one or more other immunomodulators by AD-induction in human cell lines. A suitable method for producing recombinant adenovirus is also described in Anderson, R.D., Gene Th e r ap y 7 : 1 〇 3 4 - 1 0 3 8 (200). A suitable method for the recombinant adenovirus to enter a host cell is described in Komita, Η· et al., Cancer Gene Therapy 1 6: 883-891 (2009) o to contain the hi L-12 transgene and one or more other immunomodulators and Adenoviruses from the Rheo Switch® Treatment System (RTS) transduce autologous dendritic cells. -234- 201207108 Dendritic cell lines derived from human individuals are transduced in vitro and injected into tumors. The viability, purity (usually &gt; 80% of cells showing DC phenotype), sterility, mycoplasma and endotoxin characteristics of the DC will be tested prior to viral transduction. After viral transduction, the cells are repeatedly washed to remove any unabsorbed virus. The residual virus content in the supernatant from the last rinse was detected by PCR. Since the DC line is transduced by an adenoviral vector (non-integrating virus) in vitro, and the lifetime of DC in the intratumoral injection followed by movement into the draining lymph node is short, it is not expected that the virus DN A will be embedded in any non-target 〇 cell. The method used to perform adenoviral transduction of DC is expected to produce 80 to 90% of transduction and is considered to be very efficient. PBMC were collected by white blood cell separation: subjects were subjected to a standard 90 to 120 minute white blood cell separation to the UPCI clinic for separation. The step of white blood cell separation is to draw blood from the arm vein; the blood passes through a centrifuge (cell separator) where the blood components are separated and one or more components are removed; the remaining components are returned to the same or the other arm of the individual. vein. No more than 15% of the blood is taken at any time through the cell separator

U body total blood volume. In a cell separator, blood is divided into plasma, platelets, white blood cells, and red blood cells. White blood cells (WBC) are removed and all other ingredients return to the individual's circulation. This step will try to utilize two peripheral vein (IV) catheters. If not feasible, a central venous catheter may be needed. Individuals must be sterilized by a physician before routine white blood cell separation and routinely monitor signs of life (including blood pressure). Treatment: After collection, the leukapack is manually delivered to the CPL and immediately subjected to centrifugal elutriation in ELUTRAtm. This is a closed system validated for clinical use by -235- 201207108. Collection of mononuclear ball fractions After collection and establishment of cell viability, cells were transferred to Aastrom boxes in the presence of IL-4 and GM-CSF for 6 days. All treatment and washing steps are carried out under sterile conditions. Initial vaccination: Mononuclear spheres collected from a single leukapack were counted under trypan blue staining to determine the number of viable cells. The purity of the mononuclear sphere was analyzed by flow cytometry. According to SOP-CPL-0166, the mononuclear spheres were resuspended in IX IU/ml of IL-4 and 1,000 IU/ml of GM-CSF at 5X106 to ΙΟχΙΟ6 cells/ml without serum and without Antibiotic CellGenix medium and placed in the Aastrom box. A minimum volume of 50 ml and a minimum number of cells are required for inoculation. Culture: The Aastrom cartridge is placed in an incubator for the Replicell system, a fully sealed, CGMP-compliant automated culture device for immature DCs.

Immature DC collection: On day 6, the Aastrom cassette was removed from the incubator to collect immature D C. The cells were collected by centrifugation at 1,500 rpm, washed in C e 11 G eni X medium, counted by trypan blue staining, and examined for morphological and phenotypic characteristics 〇 viability: this characteristic was carried out in the presence of trypan blue Hematocrit cell count determination. Typically, &gt; 95% of the collected cells are viable cells, i.e., the trypan blue dye is eliminated. If the viability is less than 70%, the immature DC will be discarded. Phenotypic analysis: cells produced in culture are microscopically observed on a hemocytometer, and preliminary identification calculations are obtained using trypan blue dye (DC and lymph- 236 - 201207108 cells). The identification calculation was confirmed by flow cytometry 'gating to DCs and lymphocytes, and using the high forward and side scatter characteristics of immature DCs as their identification criteria. Immature DCs typically contain &gt; 80% of cells with dendritic cell morphology and a DC phenotype. IL-12p70 titer analysis: It is well understood that mature DCs (mDCs) have the ability to produce IL-12p70 either spontaneously or upon activation by CD40L, with or without the addition of an innate immune signal (e.g., LPS). The standardized IL-12p7〇 production analysis has recently been established for use in small samples or large quantities of DC vaccines produced under a variety of conditions. The current potency analysis consists of two distinct steps involving the co-cultivation of reactive DCs with J58 8 lymphoma cells stably transfected with human CD40 ligand genes as stimulators. The second step involved the detection of supernatant from these co-cultured cells and analysis of the amount of IL-12p70 secreted by the J55 8/CD40L +/- LPS stimulated DC in the Luminex system. This potency analysis has an inter-group coefficient of variation of 18.5% (n = 30) and a broad dynamic range, which helps to evaluate the various DC products with widely varying IL-12P7 0 production. The normal range of analysis for DC preparations prepared using mononuclear spheres from 13 normal donors ranged from 8 to 999 pg/ml with an average enthalpy of 270 pg/ml. Dendritic cell preparation and release standards Each batch of in vitro prepared dendritic cells are examined for microbial contaminants (aerobic and anaerobic bacteria, fungi and mycobacteria) and endotoxin, and phenotype and functionality are determined. feature. All dendritic cells that will be injected into an individual are fresh and cryopreserved cells. -237- 201207108 DC Quality Assurance Test: The DC prepared as described above was tested for sterility, viability, purity, potency and stability. The release standards for cellular products have been established and strictly adhered to. Survivability: Cells produced in culture were microscopically observed on a hemocytometer, and differential calculations (DC and lymphocytes) were obtained using trypan blue dye. This count provides the percentage of viable cells in the test culture. More than 70% of the cells showed survival of the trypan blue exclusion and at least 70% of the cells exhibited the HLA-DR and CD86 lines of the single nuclear-derived DC marker required for release criteria. Other markers can be included for analysis, such as detecting CD83 and CCR7 in DC maturation status, and detecting CD3 and CD19 in lymphocyte sweating. Purity: Two-color flow cytometry analysis of cells stained with FITC- and PE-conjugated monoclonal antibodies was used to determine surface antigens identified as morphologically characterized by DC and lacking mononuclear and T and B cell antigens. DC group. In the case of vaccine preparation, the DC produced must exhibit HLA-DR and CD86 and cannot express CD3, CD19 or CD14. The standard considered to be mDC is that cells must exhibit CD83 + and CCR7+. Potency: To define DC titer measurements, we determined their ability to produce IL-12p70 as described above. Sterility: DCs were tested in bacteria (aerobic and anaerobic) and fungal cultures using the BD Bactec system (Becton Dickenson, Inc., Sparks, MD) at the Microbiology Laboratory at the Pittsburgh Medical Center. The end result of the microbial culture can be obtained after 14 days. Gram staining is performed prior to release of DC for vaccine use and must be free of microbes. -238- 201207108 IMCPL tested the mycobacteria using the Gen-Probe Mycobacterium Tissue Culture Rapid Test System (Gen-Probe, San Diego, CA), which is based on nucleic acid hybridization techniques. The endotoxin test was performed using pyriform cell lysate pyrogen detection (Bio Whittaker, Walkerville, MD). Endotoxin tests were performed on cell cultures while collecting cells and before release of the final product. The acceptable amount of endotoxin is &lt;5 EU/kg body weight. Dendritic cells that have not been transduced and transduced will be cryopreserved for future analysis. It is expected that all transduced cells will exhibit this transgene. More than 80% of DCs are expected to be transposed. The preparation will be biologically active as the native coding sequence is still maintained in the transgene. The virus-transduced DCs injected into the tumor are DCs with an immature phenotype and do not exhibit IL-12 and one or more other immunomodulators before they mature, so at this stage, IL-12 and one The performance of a variety of other immunomodulators is primarily derived from the transgene. Since the expression of IL-12 and one or more other immunomodulator transgenes is induced in a dose-dependent manner by the small molecule activating drug RG- 1 1 5 93 2, it is possible to control the transgene expression in the transduced DC to the desired level. The amount. A small portion of transduced DCs prepared for administration to a human subject can be tested in vitro for activation drug-dependent induction of IL-1 2 and the performance of one or more other immunomodulators. The performance of IL-12 and one or more other immunomodulators can be detected by ELISA with a sensitivity of 4 Ng/ml. It is expected that the in vitro induction of IL-12 and one or more other immunomodulators of the vector transduction cells used in the assay will produce approximately 500 ng of IL-1 2 per 1 〇6 cells and one or more other immunomodulations within 24 hours. Agent, determined by ELIS A. In a preclinical trial using a mouse melanoma model, -239-201207108 intratumoral injection of 106 or more transduced DCs showed efficacy. However, it is expected that the required intratumoral injection amount may show a therapeutic effect at a lower amount, so the 5 C 107 transduced D C is used as a starting amount to determine whether the required amount is lower or higher. For example, in vitro, human and mouse cell lines and primary dendritic cells transduced with recombinant adenoviral vectors carrying genes for IL12 and one or more other immunomodulators are shown in a dose-dependent manner. Induction of IL12 performance in response to activation of the drug. 6.3. Modulators for activating drugs The activating drugs used herein are prepared in any of the following preparations: (1) 100% caprylic acid decanoic acid polyethylene glycol glyceride (1^1?^5〇1); (2) Listerine Flavor Labrasol (Latitude Pharmaceuticals), which contains (a) menthol, (b) thymol, (alcohol, (d) aspartame, (e) saccharin Sodium, (t) citric acid, (g) mint flavor, (h) cream flavor, (i) polyglycol phthalate (labrasol); (3) Miglyol 812 and phospholipon 90G (Latitude, USA) Pharmaceuticals)); or (4) Miglyol 812, phospholipon 90G and Vitamin E Polyglycolate (Latitude Pharmaceuticals). Although it is conceivable to use various concentrations and specific procedures, a therapeutic disease-240 - 201207108 The case will include patients receiving intratumoral injection of 5 χ 10 7 of autologous dendritic cells (AdDC) and oral activating drugs (RG-1 1 5 932) suspended in sterile saline. Dendritic cell lines engineered to express hIL under the control of RTS -12 (Human Interleukin 12) and one or more other immunomodulators Initiation of treatment Day 1 Hospitalized patient visit: Basic physiology (including signs of life, weight and ECOG status) on day 1. Collection of urine And blood was drawn for basic serum chemistry, urine analysis, and hematology (safety check). Each subject was immediately after meal, approximately 3 hours prior to intratumoral injection of in vitro engineered dendritic cells. Administration of activating drugs (Group 1 - 0.01 mg / kg, 0.3 mg / kg, 1.0 mg / kg and 3 mg / kg). At specific time intervals on day 1 (before administration, 0.5, 1, 1.5 after AD administration) , 2, 4, 6, 8, 12, 16 and 24 hours) blood was drawn to assess the pharmacokinetics of a single dose of the activating drug and its major metabolites. Each subject received a single intratumoral injection of 5 χ Adenovirus-transduced autologous dendritic cells of 107 cells engineered to express hlL-12 and one or more other immunomodulators under the control of RTS. Carefully monitored subjects Local injection site reactions and / or allergic reactions. 2 to 14 Day Hospitalized Patient Visit: Days 2 to 14 'Each Subject was given active drug immediately after meal. Signs of life and adverse reactions were recorded daily on Days 2 to 14. On Day 4 ± 24 hours' Fifty percent of the subjects obtained live samples of tumors and/or draining lymph nodes to measure hIL-12 and cellular immune responses. Body weight was measured on the 8th day. Day 8 - 241 - 201207108 For 24 hours, subjects who did not obtain a live sample from day 4 obtained a live sample of tumor and/or draining lymph nodes to measure h 1L · 1 2 and one or more other immunomodulators and cells. Sexual immune response. Blood was drawn 24 hours on day 4 and day 8 to detect potential antibodies and cellular immune responses against adenovirus and/or RTS components. Serum interleukin analysis was also performed' to see if treatment with this hIL-12 and one or more other immunomodulator transgenes affected the performance of other interleukins. Urine and blood were collected on day 8 for basic serum chemistry, urine analysis, and hematology (safety check). Blood was drawn at specific time intervals on day 8 (before administration, 0. 5, 2, 4, 6, 8, 12, 16 and 24 hours after AD administration) to assess the stability of the activated drug and its major metabolites. State pharmacokinetics / ADME. Day 14 Hospitalized Patient Visit: On Day 14, each subject was given an active drug immediately after meal. Each subject received a physical examination (including signs of life, height, weight, and ECOG status). Urine and blood are collected for serum chemistry, urine analysis, and hematology (safety check). Blood was drawn on day 14 ± 24 hours to detect potential antibody and cellular immune responses against adenovirus and/or RTS components. Serum interleukin analysis was also performed to see if the performance of other interleukins was affected. Subjects' blood was drawn at specific hospital and outpatient visits to measure potential antibodies and cellular immune responses against adenovirus and RTS components. Blood was obtained for basic serum interleukin analysis. The AdVeGFP Infectious Blocking Test is used to detect antibody responses to adenoviral vectors (Gambotto, Robins et al_2004). The antibody response to the RTS component will be tested by Western blot and/or ELIS A using serum from the patient and the RTS protein produced by the expression vector at -242-201207108. In addition, multiplexed interleukin assays will be performed in serum from Luminex to detect IL-12, IFN-γ, IP-10 and other Th1/Th2 interleukins such as IL-2, TNF- α, IL-4, IL-5 and IL-10. These antibodies and interleukin assays require approximately 10 milliliters of blood. Potential antibodies and cellular immune responses to anti-adenovirus and/or RTS components: The blood of the subject is taken at specific hospital and outpatient visits to assess the potential antibodies against adenovirus and RTS components and tumor antigens And cellular plague-free response. The AdVeGFP infectious blocking assay is used to detect antibody responses to adenoviral vectors (Nwanegbo et al. 2004). The antibody response to the RTS component will be tested by Western blot and/or ELISA using serum from the patient and the RTS protein produced by the expression vector. In addition, multiplexed interleukin assays will be performed in serum by Luminex to detect 11^-12, IFN-γ, IP-10 and other Th1/Th2 interleukins such as IL-2, TNF -α, IL-4, IL-5 and IL-10. These antibodies and interleukins require 0 to about 10 ml of blood. The cellular immune response test uses approximately 50 to 60 milliliters of blood from which CD4 and CD8 T cell subsets are isolated. The isolated T cells will be mixed with autologous DC transduced with empty AdV vector, AdV-RTS or AdV-RTS-hlL 12-immunomodulator vector in the ELISPOT assay to detect AdV- and RTS-derived antigens. IFN-γ produced by activated T cells, if any. Similar assays will be performed using tumor cells as described above and/or DCs expressing shared melanoma antigens to detect early immune responses to tumors. Other test solutions can be used as needed. -243- 201207108 Pregnancy test: Women who are pregnant may have a urine pregnancy test prior to the screening visit and the first hospitalized patient visit during the retreatment period. The test is performed at least 72, 48, 24 or 12 hours prior to administration of the activating drug during the initial treatment and during all retreatments. If the urine pregnancy test is positive, it is confirmed by a serum pregnancy test. If the pregnancy is confirmed, the subject will not be able to participate in the trial or continue to participate in the retreatment period. Pregnancy testing can be done as much as possible. Drug use consultation: Before the first hospitalized patient visit in the screening and retreatment period, each subject will be required to provide a list of medications to determine the adverse effects of medications during the trial and during the follow-up period. Any possible relationship. Re-treatment criteria: Subjects will be considered for re-treatment if they are tolerant to previous AdDC vaccination without restrictive adverse effects and do not show progression or worsening symptoms at the time of possible retreatment. If the program host and the treating physician believe that the combination of additional intratumoral injection of AdD C and the 14-day continuous activation drug (the highest tolerated dose of the first study generation) may have clinical benefit, the name may be The trial provides retreatment if the following criteria are met: 1 no limiting toxicity, 2 clinical or subjective signs of stable or showing improvement in the subject's disease, and 3 no adenoviral components present in the Rheo Switch® treatment system Antibody or cellular immune response. Assessment of transgenic function and immunological effects: tumors were obtained during the experimental screening period (Day-12 to Day -7), Days 4, 8 and 14 and the first -244-201207108 months of the follow-up period And perfusion of the relevant draining lymph nodes or excision of living samples for in vivo assessment of transgenic and cellular immune responses of hIL-12 and one or more other immunomodulators. On the -12th day to the 7th day and the 14th day of the retreatment period, a fine needle aspiration biopsy sample of the tumor and related draining lymph nodes was obtained for the transgenic performance of hIL-12 and one or more other immunomodulators. In vivo assessment of cellular immune responses. Biopsy samples will be assessed using standard light microscopy and immunohistochemistry to detect cellular infiltration of T cells in tumors and draining lymph nodes. Biopsy sample sections will be interpreted by pathologists who do not know the background of the test patient. In order to distinguish between endogenous IL-1 2 and IL-12 expressed by the induced DCs in tumors and draining lymph nodes, RT-PCR of amplified RNA and appropriately designed primers will be used. Blood was drawn from the first month of the follow-up period and the -12th to 7th day, the 8th day and the 14th day of the retreatment period during the test screening period, on the 4th day, the 8th day and the 14th day. Interleukin analysis. Serum interleukin analysis was performed to see if the treatment of the hIL-12 transgene affects the performance of other interleukins. The multiplexed interleukin assay will be performed in serum by Luminex to detect IL-12, IFN-γ 'IP-10 and other Th1/Th2 interleukins such as IL-2, TNF-α, IL-4, IL-5 and IL-10. These antibodies and interleukin assays require approximately 10 milliliters of blood. Single dose and steady state pharmacokinetics of activated drugs: specific time interval on the first day of the test (0.5, 1, 1.5, 2, 4, 6, 8, 1, 2, 16 and 6 before and after the administration) 2 4 hours) blood was drawn to evaluate the single-dose pharmacokinetics of the activated drug and its major metabolites, and blood was drawn on the 8th day of the test to measure the steady state pharmacokinetics of the activated drug and its major metabolites -245 - 201207108 /ADME. Plasma was assessed by HPLC to obtain the following stable state pharmacokinetic endpoints for activated drugs and major metabolites: Cmax (highest plasma concentration), Tmax (to maximum plasma concentration time), Ctrough (minimum plasma concentration, calculated as 〇 and 24 hours) The mean concentration of 値), C24h (24-hour plasma concentration), AUC24h (area under the plasma concentration time curve from 0 to 24 hours), Ke (apparent excretion rate), and T112 (apparent half-life). It is to be understood that the scope of the present invention is not intended to be limited by the scope of the present invention. Example 3 A rationally selected, modular combination of genetic components can be rapidly assembled into a DN A expression construct via the application of a combined transgenic technique such as ULTRA VEC TORTM. In order to confirm that the genetic components that individually affect transcription, post-transcription, translation, and post-translational processing will affect gene expression together after collection, RheoSwitch technology and arbitrarily selected artificial 5'UTR, various 3'Reg + poly(A) signals are used. The combination of (SV40 and hGH), signal peptide (TNF-α and IL-2) and codon-optimized (+ /-) design to regulate transcription and increase the ability of cells to produce and secrete TNF-α. Figure 11 shows the control elements used, and shows the various module elements with special restriction sites on both sides to provide a precise combination of module combinations. The pool of modules is performed in the context of a DNA backbone designed to accept synthetic genes. An example of a ULTRA VECTORtm skeleton engineered for adenovirus packaging is shown in Figure 12. The modular design of the modular module and the adenovirus therapeutic agent - 246 - 201207108 deliver good binding because it allows the use of small regulatory sequences that are shorter than the native regulatory sequences. In vitro evaluation module combination Table 7. Matrix carrier 5 producing η test vectors and DNA combinations, UTR signal peptide ORF (CDS) 3, Reg Figure 43318 (17) TNFwt (1) TNFwtUV (4) TNFwtUV (6) SV40e + pA (8) Figure 13 43319 (18) TNFwt (1) TNFOptUV(l,2)(5) TNFoptUV (7) SV40e + pA (8) Figure 14 43320(19) TNFwt (1) IL-2optUV(3) TNFoptUV (7) SV40e + pA (8) Figure 15 43321 (20) 5U2 (2) TTNFwtUV (4) TNFwtUV (6) SV40e + pA (8) Figure 16 43322 (21) 5U2 (2) TNFOptUV(l,2)(5) TNFoptUV (7) SV40e ten pA (8) Figure 17 43323 (22) 5U2 ( 2) IL-2optUV(3) TNFoptUV (7) SV40e Ten pA (8) Figure 18 43324 (23) TNFwt (1) TNFwtUV (4) TNFwtUV (6) hGH + pA (9) Figure 19 43325 (24) TNFwt ( 1) TNFOptUV(l,2)(5) TNFoptUV (7) hGH + pA (9) Figure 20 43326 (25) TNFwt (1) IL-2optUV(3) TNFoptUV (7) hGH+ pA (9) Figure 21 43327 (26) 5U2 (2) TNFwtUV (4) TNFwtUV (6) hGH+ pA (9) Figure 22 43329 (28) 5U2 (2) IL-2optUV(3) TNFoptUV (7) hGH+ pA (9) Figure 24 Vector transiently transfected to ΗΕΚ2 93 Τ Cell lines to assess which combination of modules results in increased TNF-[alpha] production. To induce the expression of TNF-a, a ligand or vehicle is administered to control the cells. The supernatant was collected and the amount of TNF-α was measured by ELIS A. In order to set a base similar to the wild type, the vector 43 3 18 contains UV in accordance with the wild type TNF-α 5' UTR, the signal peptide, the coding sequence and SV40pA 3, Reg. Individually, the module of the TNF-α signal peptide or mature protein coding sequence was changed to 〇pt(l,2) codon optimization resulting in a gradual increase in protein secretion (vectors 43319, 43320). In addition, the secretion amount (vector 43 3 22, 43 3 23 ) was further provided by taking the -247-201207108 generation TNF-wt 5WTR with the 5U2 5'UTR module. The highest secretion of TNF-α is when wild-type 5, UTR, signal peptide and coding sequence modules are replaced by 5U2, IL2 and TNFOptUV modules, respectively (vector 43329) ° To confirm that the increase in secretion of TNF-α is independent of cell type, One of the experimental vectors and two control vectors were transfected into CH0-K1 cells (see Figure 27). Vector 43534 (Figure 26) and vector 43533 (Figure 25) contained the wild type TNF-[alpha] module as a control. Vector 43 534 is composed of a wild-type TNF-α sequence and does not contain an Ultra Vector collection fulcrum. It is worth noting that the presence of the UltraVector set fulcrum does not adversely affect the production and secretion of THF-α. In this data, we also demonstrated that replacement of TNF-α wild-type 3'Reg with a polyA module containing SV40e or hGH resulted in increased secretion of TNF-α. The maximal TNF-a production was achieved by a combination of 5U2, IL2 signal peptide, TNFOptUV and hGHpA. Data from CHO-K1 cells showed that the modules gradually increased with the 5U2, IL-2 signal peptide and TNFOptUV coding sequences in SWTR. Interestingly, the magnitude of the increase is slightly different in the two cell types. This means that although the modules function similarly in various cell types, physiological differences in specific cell or tissue types may affect the extent of module substitution effects. Extending the combinatorial matrix to include more modules in each category may allow for the best combination to be found by cell type or test tissue. Additional module examples that may be included in a larger matrix are included in SEQ ID NOs: 41-46. Example 4: Evaluation of therapeutic candidates in animal models In order to show the effectiveness of inducible-optimized TNF-a constructs for the treatment of cancers such as prostaglandin-248-201207108 adenocarcinoma or head and neck cancer, head cancer mice can be used for the disease. model. The Smad4 single gene knockout has been shown to produce a spontaneous model of malignant human head and neck cancer squamous cell carcinoma (HNSCC) (PMID: 19841536). In the absence of this mouse strain, human-derived HNS C C tumor cells can be implanted into nude mice. After the tumor is established, the optimized TNF-[alpha] construct can be introduced into the tumor along with the adenovirus. Various doses of the ligand can be administered to the mouse to modulate the amount of optimized TNF-a produced. Tumor burden was measured and tumor necrosis was assessed to identify possible therapeutic candidates from the optimized TNF-a construct. Example 5: Example of therapeutic embodiment The engineered TNF-a transgene was administered to a patient by intratumoral injection of a non-replicating adenovirus DN A vector. This gene program encodes a codon-optimized mature mammalian interleukin that is fused to a codon-optimized IL-2 signal peptide. Next, wild-type TNF-a 5'UTR and SV40 3· Reg + poly(A) signals were added to both sides of the transgenic cds (IL-2 SP + TNF-a), and the expression was performed by RheoSwitch technology. Careful "directed" dose of activated ligand control (ie, DNA normalized in vector VVN-43 320, see Figure 15). Preliminary data indicate that this combination of DNA elements produces the highest likelihood of TNF-a secretion induction while still providing control over his performance of "tightening" and "not relaxing." That is to say, the transformation of the transgenic configuration is not induced, and the performance of the basic amount is maintained at a low level, and it is relatively impossible to exhibit an uncontrolled and off-target effect on the patient. In an alternative embodiment of the invention, the engineered TNF-a trans-249-201207108 gene is administered to a patient via an adenovirus in a modular DN A configuration similar to vector VVN-43329 (see Figure 24). It simultaneously exhibits high substrate performance and highest transgenic performance. This DNA uses artificially engineered 5'U2 and hGH poly(A) signals, as well as intact mammalian codon optimization and IL-2 signal peptides. Control of systemic toxicity in patients is achieved by intratumoral injection of adenovirus using low MOI. To a lesser extent, performance and transient control can also be regulated by the RheoSwitch activation ligand. Additional control of the distribution of this gene product can be achieved by incorporating tissue-specific miRNA response elements that inhibit off-target expression in vital organs or enhance the affinity via artificially engineered adenoviral nucleocapsids. In an additional embodiment of the invention, the engineered TNF-[alpha] transgenic line is similar to the DNA in VVN-43 3 2 8 (Figure 23), which is predicted to confer the artificial mRNA height via a 5' U2 element stability. However, this construct does not utilize the IL-2 signal peptide to enhance secretion, and it maintains the wild-type sequence from the DNA encoding the Ν' end of TNF-α. For unknown reasons, high-volume secretion of artificial TNF-ct can be clearly confirmed to be unfavorable for patients regardless of the prognosis of the content. However, the natural mechanism of interleukins via the metalloproteinase "shedding" may be limited by this factor. The tumor environment limits the toxicity of the patient. If the natural shedding of exogenous TNF-α still shows a deviation from the target, the extracellular domain stem can be truncated by mutation to inhibit the dissolution of the native protease, and the factor will be contacted via cell-cells. The second type of transmembrane protein is activated to activate. Alternatively, a transgene similar to the construct described by vector VVN-43 3 2 8 can encode a constitutively expressed TNF-α having a mutant stem cell containing a site for cleavage by an exogenous protease. - 201207108 Outer domain. In addition, the source protease will in turn be placed under the promoter element of the Rheo Switch technology control and will only be expressed in the presence of the activating ligand. Thus, cleavage of TNF-[alpha] and in vivo solubility (but not surface expression) will be controlled by modular transgenic elements. Example 6: Anti-tumor effect of Ad-RTS-IL-12 The anti-tumor effect of Ad-RTS-IL-12 has been applied to a series of mouse tumors of melanoma, colorectal cancer, pancreatic cancer, breast cancer, lung cancer and kidney cancer. The model is evaluated. An exemplary dose response experiment is shown below. The immunocompetent C57bl/6 mother (6 to 8 weeks old) was subcutaneously administered to B16F0 mouse melanoma cells. Mice were grouped 11 days after tumor cell inoculation when there were significant macroscopic tumor nodules (mean tumor volume approximately 40 mm3), 5 animals per group. A total of 9 groups: control saline treatment group, activated ligand treatment group, only eight (1-11 Ding 3-111112 (16610 mouth) group and another 6 groups with different doses of carrier human &lt;1-foot 78-11111 ^-12 (167, 168, 169, 569, 1610, 5610 virions) plus activation vehicle treatment. The mice in the ligand group were given the carrier one day before the 2018 Teklad Global 18% protein RodeiU Diet (Harlan Lab (Harlan Lab oratories)) 1 mg/kg activating ligand (looo mg ligand/kg feed) is provided in the feed. Control mice provide 2〇18

Teklad Global 18% Protein Rodent Diet. On day 12, a single dose of 100 microliters of PBS in Ad-RTS-mIL12M was injected into the tumor. Tumor volume and body weight were measured using vernier calipers and scales every 2 to 3 days, and animals were followed until the control tumors reached 2000 mm3. The data was uploaded to the study log animal test software. -251 - 201207108 As shown in Figure 31, a significant anti-tumor effect occurred in the Ad-RTS-IL-12 dose above the iee8 vp (range 73 to 99%). The lowest test dose of lee7 vp Ad-RTS-IL-12 did not show an anti-tumor effect. Ad-RTS-IL-12, a high dose of leelO vp, was not effective in the absence of activating ligand, indicating that a combination of both Ad-RTS-IL-12 and an activating ligand is required. The treatment of the ligand itself is also ineffective. Therefore, this assay shows that this potent anti-tumor effect is mediated by a combination of Ad-RTS-IL-12 and an activating ligand. Body weight analysis is shown in Figure 32. Animals treated with the highest dose (5 eel 0 vp) of Ad-RTS-IL-12 showed temporary weight loss on day 19 but recovered on day 26. Animals in the other treatment groups did not show any clear dose-response relationship and only minor changes in body weight gain were observed. Example 7: Efficacy of Ad-RTS-IL-12 in the Lewis lung cancer model Immunocompetent C57b/6 mothers 6 to 8 weeks old were inoculated subcutaneously (s.c.) with mouse Lewis lung cancer cells (LLC). Five days after cell inoculation, mice were randomly assigned to a total of 4 treatment groups and a control group (n = 5), respectively: no treatment group (control), only activated ligand group (RG-1 1 5932), Only the Ad-RTS-mIL12 group and the Ad-RTS-mIL12 plus activation ligand group. The group receiving the activated ligand (L) was fed ad libitum with 2018 Teklad Global 18% Protein Rodent Diet (Harlan Laboratories) feed mixed with the activating ligand (1 000 mg/kg feed). Continue to feed general feeds in groups that receive only Ad-RTS-mIL12 or no treatment. Treatment begins when the tumor reaches 28 ± 6 mm 3 . Ad-RTS-mIL12 (lelO vp/100 microliters of PBS) was administered to mice via intratumoral (i.t.) injection on days 6, 9, and 13 after tumor cell inoculation -252-201207108. The activated ligand feed (L) was administered to the mice 24 hours prior to vehicle administration. The tumor volume and body weight of each mouse were measured three times a week using a vernier caliper and scale until the end of the experiment. The test was terminated when the tumor volume of the mouse was &gt; 1 200 mm 3 . The data was uploaded to the Study Log animal test software. The tumor volume after treatment is shown in Figure 33A. Lewis lung tumor-bearing mice in the control and activated-only ligand (L) groups showed approximately similar tumor growth kinetics. Only three doses of the Ad-RTS-mIL 12 group resulted in intermediate tumor growth. Importantly, Ad-RTS-mIL12 plus activating ligand (L) produced significant tumor growth inhibition (78%) compared to the control group. This data shows that Ad-RTS-mIL12 inhibits the growth of Lewis lung tumors in the presence of activating ligands. Body weight was monitored as an indicator of toxicity. No significant weight loss was found during the trial. Example 8: Anti-tumor effect of Ad-RTS-IL-12 in melanoma model Immunocompetent C57b/6 mothers aged 6 to 8 weeks subcutaneously (sc·) of mouse melanoma cancer cells (B16F0) Vaccination. One day after cell inoculation, mice were randomly assigned to a total of 9 groups of treatment and control groups (n = 5), respectively: no treatment group (control), only activated ligand group (L) (RG-1) 1 5 93 2), Ad-RTS-mIL12 group and Ad-RTS-mIL12 plus different doses (50, 1〇〇, 250, 500 and 1000 mg/kg) of the activated ligand group. The group receiving the activated ligand (L) was fed ad libitum with the activating ligand (1 000 mg/kg feed) 2018 Teklad Global 18% Protein Rodent Diet. Accept only Ad-RTS-mIL12 or untreated group for continuous feeding - like 2〇18

Teklad Global 18% Protein Rodent Diet chow diet feed. -253- 201207108 Treatment begins when the tumor reaches 56 ± 18 mm 3 . A single dose of 8% RTS-mIL12 (lelO vp/100 microliters of PBS) was administered to mice via intratumoral (i.t.) injection on day 13 after tumor cell inoculation. The activated ligand feed (L) was administered to the mice 24 hours prior to vehicle injection. Tumor volume and body weight per mouse were measured three times a week using a vernier caliper and a pound scale until the end of the trial. The test was terminated when the tumor volume was &gt; 2000 mm 3 . The data was uploaded to the Study Log animal test software. Tumor volume and body weight changes after treatment are shown in Figures 34A and 34B. Melanoma-bearing mice in the control and activated only ligand (L) groups showed similar aggressive tumor growth. Tumor growth kinetics showed that the activated ligand group did not have antitumor activity. Animals receiving a single dose of Ad-RTS-mIL12 (lelO vp) but without ligand observed mild tumor growth inhibition (12%) on day 26. Eight £1_11 Ding 3-11111^2 plus activating ligand (1〇 treatment resulted in tumor growth inhibition (73 to 98%) compared to control mice. Single dose of Ad-RTS-mIL12 plus 50 mg/kg activation The ligand (L) produced a significant tumor reduction compared to the control tumor. It is noted that when the dose of the activating ligand is increased to 100 to 1 000 mg/kg compared to the 50 mg/kg activating ligand, Significant anti-tumor activity (90 to 98%). This data clearly shows that eight £1-11, 3-11111^12 is effective in the melanoma model and exhibits a wide range of therapeutic activation ligand dose windows. Body weight was used as a toxicity indicator. On Days 13 and 17, there was a slight temporary weight change (&lt;5%) of the activated ligand dose of 100 mg/kg. No severe weight loss was found during the rest of the trial. No activation ligand dose-response-related changes in body weight were found. AdRTS-mIL12 treatment was well tolerated at different doses without any signs of toxicity. -254- 201207108 Example 9: Ad-RTS-IL-12 in colon cancer Anti-tumor efficacy in the model 6 to 8 weeks old immunocompetitive Balb/C The mice were subcutaneously (sc) inoculated with luciferase-stable mouse colon cancer cells (CT26LUC). Ten days after cell seeding, mice were randomly assigned to a total of 3 groups of treatment and control groups (n=5). ), the untreated group (control), the activated ligand only group (RG_1 1 5932), and the Ad-RTS-mIL12 plus activated ligand group. The group receiving the activated ligand (L) was fed ad libitum and activated. 2018 Teklad Global 18% Protein Rodent Diet (Harlan Laboratories) feed for the body (1 000 mg/kg feed). Continued feeding with untreated groups 2018 Teklad Global 18% Protein Rodent Diet chow Treatment begins when the tumor reaches 40 ± 17 mm 3 . Eight (1-11 butyl 8-mIL12 (lelO vp/100 μl PBS) is injected intratumorally (it) on days 11 and 18 after tumor cell inoculation The mice were administered to the mice. The activated ligand feed (L) was administered to the mice 24 hours prior to vehicle injection. The tumor volume and body weight of each mouse were measured three times a week using a vernier caliper and a scale until the end of the experiment. When the mouse tumor volume was &gt; The test was terminated at 2000 mm 3. The data was uploaded to St. Udy Log animal test software. Tumor volume and body weight changes after treatment are shown in Figures 35A and 35B. Colon cancer-bearing mice in the control and activated only ligand (L) groups showed similar aggressive tumor growth. Tumor growth kinetics showed that the activated ligand group did not inhibit tumor growth. The second dose of Ad-RTS-mIL12 plus activating ligand (L) resulted in complete regression and tumor growth inhibition (100%) compared to control mice. Chad Note -255- 201207108 means that Ad-RTS-mIL12 treatment resulted in five of the five animals completely free of tumors. Mice without tumor after Ad-RTS-mIL12 treatment were challenged with parental CT26LUC cells. Five Balb/c mice that had not received vaccination were also subcutaneously inoculated with CT2 6LUC as a control group. The control mice produced tumor nodules as expected. Importantly, no tumors occurred in all challenged mice 4 weeks after the challenge. This trial showed that Ad-RTS-mIL12 treatment developed strong anti-tumor immunity in an aggressive colon cancer model. Monitor body weight as an indicator of toxicity. No significant weight loss was observed during the trial. Example 10: Anti-tumor efficacy of Ad-RTS-IL-12 in pancreatic cancer model Immunocompetent C57b/6 mothers aged 6 to 8 weeks old with homologous PAN02 pancreatic cancer cells Subcutaneous (sc) inoculation of (ATCC). Six days after cell inoculation, mice were randomly assigned to 4 groups of 5 animals each, which were untreated, activated ligand only (RG-1 15932), Ad-RTS-mIL12 only, and Ad- RTS-mIL12 plus activated ligand group. The 2018 Teklad Global 18% Protein Rodent Diet (Harlan Laboratories) feed, admixed with the activated ligand (1 000 mg/kg feed), was fed ad libitum. Continue to feed 2018 Teklad Global 18°/ for groups that receive only Ad-RTS-mIL12 or no treatment. Protein Rodent Diet chow. Mice were treated with a single intratumoral (i.t.) injection of Ad_RTS-mIL12 at a dose of lelO vp/100 microliters of PBS on days 7 and 14 after tumor cell inoculation. At the beginning of vehicle treatment, the tumor volume averaged STGT mm3. The tumor volume and body weight of each mouse were measured three times per week until the test knot -256 - 201207108 bundle. The test was terminated when the tumor volume of the mouse exceeded 600 mm 3 . Since the pancreatic tumor grows very slowly, we define the termination of the trial. Tumor growth was normal in mice that were not treated. In this tumor model, microscopic tumor growth delay was noted in mice receiving either activating ligand alone or only Ad-RTS-mIL12 treatment. In contrast, tumor growth in all Ad-RTS-mIL12 treated mice was significantly inhibited (97%) compared to untreated control mice. Toxicity was measured using a vernier caliper and scale to measure body weight during the test. The body weight of the mice injected with Ad-RTS-mlL 12 showed no significant weight loss after administration, except for the temporary weight loss (&lt;5%) on days 12 to 13. In addition, no pathological behavior (tiredness, rough skin, lameness, dehydration, bow-back posture, etc.) was observed in any animal. Tumor regression was maintained until day 37 when control animals were sacrificed. The data was uploaded to the Study Log animal test software. The results are shown in Figures 36A and 36B. Example 1 1 : Anti-tumor effect of Ad-RTS-IL-12 in a breast cancer model The purpose of this test was to evaluate the efficacy of human &lt;1-11 D 3-11111^12 intratumoral treatment in a mouse breast cancer model and toxicity. BalbC mothers, 6 to 8 weeks old, were purchased from Charles River Laboratories or Harlan Laboratories (USA). Animal care and laboratory procedures are conducted in accordance with Intrexon's Institutional Animal Care and Use Committee guidelines. The mouse breast cancer (4T1) cell line was purchased from the American Type Culture Collection (ATCC) (Manassas, VA). The 4T 1 cell line was grown at the Roswell Memorial -257- 201207108 Roswell Park Memorial Institute medium (RPMI) 1 640 (ATCC, Manassas, VA). The medium was supplemented with heat-inactivated fetal bovine serum (FCS) 10% v/v, 2 mM L-glutamic acid (Atlanta Biologicals, Inc., Lawrenceville, GA), 1〇 〇 IU / ml penicillin G and 100 μg / ml streptomycin. The cells were grown at 37 ° C, 5% CO 2 . All cell lines were routinely tested and no mold was found. Immunocompetent BALB/c mothers of 6 to 8 weeks old were inoculated subcutaneously (s.c.) with homologous breast cancer (4T1) cells (le5 cells/50 microliters). Eight days after cell inoculation, mice were randomly assigned to 4 groups of 5 animals each, which were untreated, activated ligand only, Ad-RTS-mIL12 only and 1-11 m IL 1 2 plus activated ligand group. The mice receiving the activated ligand were fed ad libitum with the mixed activation ligand (1 mg/kg) of mouse feed. Only Ad-RTS-mIL12 or no treatment group was accepted. Do not continue to feed standard feed (Harlan Laboratories, USA). The activation system is administered via a custom feed produced by Harlan Teklad (Harmon's Habitat Feed Division) at a rate of 1 kg and fed the control animals. The feed contains 1 〇 0 mg of activating ligand. The mice received a dose of 1 e 1 〇vp/100 μl of PBS Ad-RTS-mIL12 on days 9, 12 and 14 after tumor cell inoculation. Single intratumoral (丨丄) injection therapy. The mean tumor volume was 36 mm 3 at the start of vehicle treatment. The tumor volume and body weight of each mouse were measured three times per week until the end of the experiment. When the tumor volume of the mouse exceeded The test was terminated at 1 000 mm at 3 o'clock. 8 days after the inoculation of breast cancer 4T 1 cells, the mice were The machine was assigned to a total of 4 groups of treatment and control group (η = 5 / group), respectively, no treatment group (control), only -258-201207108 activation ligand group (L), Ad-RTS_mIL12 only and Ad-RTS -mIL12 plus activation ligand group, as shown in the following table. Breast (4T1) tumor model treatment design group N Activated ligand (L) feed General mouse feed tumors after intracellular administration of cells to start treatment of tumor volume, body weight 1 5 No Yes Monday, Wednesday and Friday 2 5 Whether Monday, Wednesday and Friday 3 5 No Yes Ad-RTS-mIL12 Days 9, 12, 14 Monday, Wednesday and Friday 4 5 Whether Ad-RTS-mIL12 No. 9 On days 12 and 14, Monday, Wednesday and Friday, when the average tumor volume reached 3 6 mm 3 , the tumor volume after treatment was as shown in Figure 39A. Only Ad-RTS-IL-12 treatment and only activation of ligands ( L) 4T1 tumor-bearing mice in the control group showed ~20% and 35% tumor growth inhibition on day 26, respectively (Fig. 39A). Importantly, three doses of Ad-RTS-mIL 1 2 plus activating ligand Significant tumor growth inhibition (82%) compared to the control group (p &lt; 0.005). This data shows that Ad-RTS-mIL12 is present in the activated ligand It exhibited potent antitumor activity in the breast cancer (4T1) model. Body weight was monitored as a toxicity indicator. No severe weight loss or death was observed during the trial (Fig. 39B). These results show direct intratumoral injection of Ad-RTS- The mILl 2 plus activating ligand is highly effective in inducing tumor regression and is safe in breast cancer models. The antitumor activity in this model was significant (P &lt; 0.005). -259-201207108 Example 12: Clinical procedure for administration of Ad-RTS-IL-12 vector without immunocytes The following is a treatment that can be used to practice the present invention in the form of Ad-RTS-IL-12 vector. A clinical trial program for m- or IV-stage malignant melanoma. The purpose of this Phase lb clinical trial was to evaluate the safety and objective response of the combination of intratumoral injection of Ad-RTS-IL-12 and 14 days of daily oral administration of activating ligands over 6 treatment cycles. Immunity and other biological activities. The Ad-RTS-IL-12 dose will be administered starting with 1 X 1011 virus particles (vp) and a 5 mg/day dose of activating ligand (Phase 1). The dose of both the viral particles and the activating ligand will then be adjusted in each repeated treatment cycle per subject according to a fixed schedule (Table 8), provided that the patient can tolerate the previous treatment cycle. The purpose of this Phase lb trial is as follows: 1. To evaluate the repeated treatment cycle of a combination of AD-RTS-IL-12 and a dose-enhancing activation ligand in a tumor-injected patient for an unresectable III C or IV Safety and tolerability of patients with malignant melanoma. 2. Obtain therapeutic indications by diagnostic CT scan (Recognition of Solid Tumor Response (REC 1ST 1.1)), PET scans, and photographs (if applicable). 3. To assess the functionality of the RheoSwitch therapeutic system (RTStm) in patients, by assessing the immune effect of AD-RTS-IL-12 in combination with an activating ligand to receive the target tumor, tumor-associated draining lymph node (if available) Close to) and cellular immune responses in peripheral circulation (especially the gene expression of IL-12 and other interleukins, the frequency of cytotoxic T lymphocytes and Tregs) -260- 201207108 and other biological activities (such as apoptosis and In the case of immune cell infiltration, 'and find out the relationship between these immune and other biological parameters and the previously activated ligand dose and tumor response. 4. Assess the extent of uptake of dendritic cells and macrophages in tumor cells and tumors from 8-11 to 3-11^12 to determine which cells take up virus regardless of the level of uptake depending on AD-RTS-IL- 12 doses. Inflammatory responses and immune responses (cytotoxic such as cytotoxic lymphocytes and Tregs, and induced interleukins) in tumors, tumor-associated draining lymph nodes (if accessible), and peripheral circulation are measured. The relationship between changes in immune and other biological parameters and the dose of AD-RTS-IL-12 and activating ligands and tumor response will be identified. 5. Evaluate the pharmacokinetic properties of the steady state phase on days 8 to 9 of each cycle in the patient subpopulation. 6. Evaluate the QT/QTc interval of the electrocardiogram of patients who will undergo PK assessment. The ECG is obtained from Holter monitoring. Indications: Stage III C (metastasis in the pathway), stage IV (Mia, Mlb or Mlc (LDH&lt; = 2xULN) malignant melanoma, with at least 4 accessible lesions. Test design: 1b Period, open-label, single-group, multi-center assessment of safety, tolerability, tumor response (RECIST 1.1), and immune and other biological effects in each of the six treatment cycles, each cycle lasting 28 days, each cycle of intratumoral injection of eight (1-11 Ding 3-11^12 and 14 days per sputum oral administration of a combination of activated ligands. The dose of 0-11 Ding 3-1!^-12 and the activating ligand will be according to Figure 1 and Table 1. Shown in all patients who can tolerate previous treatment cycles. -261 - 201207108 Test population: All ethnic males and females 'age 18 years and older' with unresectable stage III c or IV malignant melanoma ' ECOG fitness status 0 to 1, at least 4 accessible lesions (maximum diameter &lt;= 3 cm 'minimum diameter> =1 cm) or sensible tumor-associated lymph nodes (maximum diameter &lt;= 5 cm, Minimum diameter &gt; =1 · 5 cm) for intratumoral injection and biopsy. Sample size A minimum of 12 patients and a maximum of 28 patients with stage III C or IV malignant melanoma will participate in the trial. All patients in this trial will enter a single group, and each treatment cycle will be based on Figure 1 and Table 1 shows an increase in the intra-dose dose of AD-RTS-IL-12 and activated ligands, provided that the patient is able to tolerate the previous treatment cycle. Test article: Patients will receive oral activating ligands in each cycle Intratumoral injection of gene therapy (Ad-RTS-IL-12), which is engineered to exhibit inducible hIL-1 2 by dose-dependent response to activating ligands. AD-RTS-IL -12 will be prepared and frozen in a central manufacturing facility and shipped to the appropriate clinical center. All patients will receive intratumoral injections (once per cycle, up to 6 cycles, 4 weeks apart) AD-RTS- IL-1 2 (about 1.0 X 1 0 11 and 1 · 0 X 1 〇12 total viral particles per injection). Patients will also receive oral activating ligand once a day for 14 consecutive days. AD- The dose of RTS-IL-1 2 and/or activating ligand will be at the beginning of cycles 2 to 6 (see Table 8) 'at all energy In patients with tolerance to the previous treatment cycle, AD-RTS-IL-12 will be injected into different lesions in each cycle, -262 - 201207108 If the number of lesions is limited, the injection will be in order The method was performed in turn. One of the at least 4 accessible lesions was not injected and was used to assess the systemic effects of AD-RTS-IL-12. Patients will be staggered at least 24 hours apart. There will be an intratumoral injection per cycle occurring approximately 3 hours (± 30 minutes) after administration of the first dose of activating ligand. Dosage: Activated ligand: lowest dose/day: 5 mg; intermediate dose/day: 20 mg; highest dose/day: 100 mg. The activating ligand will be administered within the first 14 days of each cycle.

Ad-RTS-IL-12: Dosage · · Each injection about 1. 〇X 1011 or 1.0 X 1 0 1 2 virus particles / tumor suspended in total volume 〇. 5 ml sterile solution, the injection volume is distributed throughout the lesion, especially It is the area at the edge of the tumor. Table 8: Dosing week AD-RTS-IL_12 dose (swell injections / activated ligand mg / number of activated ligands / intratumoral) viral particles (VP) cycle daily dose (oral) period 1 l.Ox 1011 1 (Day 1) 5 14 (Days 1 to 14) 2 1.0 xlO11 K Day 1) 20 14 (Days 1 to 14) l.Ox 1011 1 (Day 1) 100 14 (1st to 14 days) 4 l.Ox 1012 1 (Day 1) 5 14 (Days 1 to 14) 5 l.Ox ΙΟ12 1 (Day 1) 20 14 (Days 1 to 14) 6 l.Ox 1012 1( Day 1) 100 14 (Days 1 to 14) Treatment with the next highest dose will not begin until the safety and tolerability of the previous treatment have been confirmed. If the MTD is defined, it will not continue to improve. Route of administration: Activated ligand: solution in soft gelatin capsules, served in the mouth for 30 minutes -263-201207108; AD-RTS-IL-12: injected to an accessible tumor on the first day of each cycle The lesion or if necessary, is injected into a tumor-associated (accessible) draining lymph node. Patient grouping method: All patients will receive treatment according to Table 8, entering a group. The safety and tolerability of all patients will be rigorously tested during or after each treatment cycle. The dose will only increase if the previous treatment cycle is well tolerated. During the trial: This trial will last approximately 28 weeks after screening for each patient. After a screening evaluation period of up to 23 days (days -30 to -7), the patient will be approved for the trial. A baseline biopsy will be performed on days -6 to 2, and a baseline cardiac function assessment will be performed on day 0 for patients assessed for PK using a continuous electrocardiogram (Hoiter monitoring). On the first day of each cycle, the approved patient will begin the trial treatment (intratumoral injection of AD-RTS-IL-12 and sequential activation of the ligand oral dose). The activated ligand treatment in each cycle will last for a total of 14 days, and the next 14 days will be cleared and observed for safety. The trial treatment consisted of 6 cycles, each maintained for 28 days, including a 14-day follow-up period. The post-treatment follow-up assessment will be performed 6 weeks after the last injection (4 weeks after the last activation of the ligand dose). The viral DN A in the blood was measured. Viral DNA will continue to be detected if viral DNA is present 6 weeks after the last injection. However, if Q-PCR from each source exhibits a second consecutive negative result, no further testing is required. Primary endpoint: -264- 201207108 Safety and tolerability will be checked by physiology (including ECOG status), QT/QTc interval in electrocardiogram (in PK patients), signs of life, serum chemistry, urinalysis, hematology and Any adverse reaction reports from the patient are evaluated. Objective responses and response rates were assessed by CT scan. Secondary endpoint: Stable state pharmacokinetics of activating ligands in 8 patient subgroups (4 patients per AD-RTS-IL-12 dose). b. The extent of inflammation, immune response (cytotoxic such as cytotoxic lymphocytes and Tregs, and induced interleukins) in tumors, tumor-associated draining lymph nodes (if accessible), and peripheral circulation due to treatment. c. Changes in immune and other biological parameters associated with AD-RTS-IL-12 and activating ligand doses and tumor response. d. The efficacy of PET scanning and photo evaluation. e · Long-term tracking will last up to 5 years. The program host contacts patients with annual inclusion criteria: a. All races, men or women over the age of 18; b. Unremovable III C (transfer in the pathway) or stage IV melanoma (Mia, Mlb , Mlc and LDH &lt;= 2x ULN), melanoma from any tumor thickness of the original skin, mucosa or underarm, or from unknown origin; c. at least 4 accessible non-visceral lesions (maximum diameter &lt;; = 3 cm 'min > =1 cm) or the tumor-associated lymph nodes (maximum diameter &lt;= 5 cm 'min > 1 · 5 cm) for intratumoral injection or biopsy. At least -265- 201207108 A lesion will not be injected. d. ECOG stomach g @ state or 1; e. Patients without visible brain metastases, assessed by contrast-enhanced MRI scan at screening or within 30 days prior to the trial; f. appropriate baseline hematology and organ function 'The number of laboratories within 30 days before the treatment treatment and the repeated treatment cycles and the dose of the activated ligand were evaluated as follows: hemoglobin &gt; = 1 g/l, particle ball &gt; 2500 / mm 3, Lymphocytes &gt; 1000/mm 3 , platelets &gt; 1 〇〇, 〇〇〇 / mm 3 , serum creatinine &lt; 1 · 5 X ULN, AST, ALT, alkaline phosphatase &lt; 2.5 X ULN, LDH &lt;;= 2 X ULN, serum bilirubin &lt; 1. 5 X ULN, absolute neutrophil &gt; 500 / mm 3 ; g. Project host believes that expected survival is at least about 6 months (mainly Assessment by physical status) h. Women must have menopause or surgical sterilization or effective contraception; men who have not undergone surgical sterilization and have not had menopause or surgical sterilization must implement effective contraception; i. Normal coagulation parameters measured by PT/PTT j. Sign the IRB approved subject consent form. Exclusion criteria: a. Initiatives requiring specific treatment, acute viral, bacterial or fungal infections; b. HIV infection, as it may not be able to elicit an effective immune response; c. Requires steroids (&gt; 10 mg of prednisone ( Active prednisone) or other immunosuppressive agents for the treatment of autoimmune diseases; -266- 201207108 d. Patients with detectable brain metastases at screening (or within 30 days prior to the trial), with contrast enhancement MRI scan evaluation; e. lesions > 3 cm (LD) or sensible tumor-associated lymph nodes > 5 cm (LD) patients; f. hemoglobin &lt; 1 g / g patients; g Dirty metastases or other distant metastases in stage IV, if LDH &gt; 2 X ULN ; h. Patients who have received AD-RTS-IL-12 or activated ligand therapy; i. have received intratumoral genes Patients with therapy; j. recipients of organ xenografts; k. other clinically active malignant diseases, except for other skin cancers; l. from previous chemotherapy, hormonal therapy, radiation therapy, immunotherapy or any first line The treatment is completed less than 30 days (in the first dose Before the test); m. Clinically significant cerebrovascular disease; η·倂Severe cardiac insufficiency (New York Heart Association classification III or IV) or coronary artery disease or its history; 〇· may be considered an unacceptable anesthesia Or acute medical condition at the risk of surgery such as ischemic heart disease or lung disease; 倂. 倂 blood or coagulopathy or a history thereof; q. use immunosuppressive therapy such as corticosteroids (&gt; 10 mg prednisone or Equivalent) and cyclosporin A; r. 硏 use deliberate treatment, or receive any deliberate treatment within the past 30 days (before the first dose of the test drug); -267- 201207108 s Use drugs that are metabolized via the CYP45 0 3 A4 pathway; t. breast-feeding or pregnant women; u. patients with a history of allergies to any component of the product, such as benzoic acid associated with activating ligands, which contain two benzene rings v. Any medical or mental condition that is considered by the program host to be unacceptable to reduce the safety or delivery of the proposed treatment, or to obtain a voluntary consent form. Statistical Methods: The objective response (CR + PR) was assessed based on the size of the injected and non-injected tumor lesions and the sensible tumor-associated lymph nodes obtained by CT scan [Using the Solid Tumor Response Assessment Criteria (RECIS T 1.1)]. PET scans and/or photographs will be used to assess metabolic activity or size (skin lesions), respectively. The primary analysis of OS and ORR will include the confidence interval and will be performed when the number of samples reaches 12, 16 '20, 24 patients and 6 weeks after the last patient is treated. Demographics, immunology, and biological activity data, as well as safety parameters including adverse reaction frequency and laboratory numbers, will be descriptively analyzed at the end of the follow-up period. The results will be summarized into a table, a picture and a list of patient results. Descriptive statistics including mean 中, median, standard deviation, and histograms will be used to summarize consecutive numbers. Frequency calculations will be used for class variables, including objective tumor responses. Immunity and biological activity will be associated with anti-tumor effects. These statistics will be provided in layers (tumor lesion size: &lt; = 1 cm maximum diameter [LD], &gt; 1 cm LD; size of DLN involved: &lt;= 3 cm LD, &gt; 3 -268 - 201207108 cm LD ; lesion location: visceral, non-visceral; lesion injection status: injection, not injected). Statistical analysis will be performed at each treatment cycle and after the end of the trial. In the case of holistic analysis, observations can be combined across layers but not across cycles. Compliance: This trial was conducted in accordance with the current Good Clinical Practice (eGCP). References Abdalla, 2007.

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Huang, H. and Xiang, J. (2004) Synergistic effect of lymphotactin and interferon gamma-inducible protein-10 transgene expression in T-cell localization and adoptive T-cell therapy of tumors. Int. J. Cancer. 109, 817- 825. [Simplified Schematic] Figure 1 shows the plastid gene map of the bicistronic transcript-regulated promoter expression system encoding hIL-12. Figure 2 shows a plastid gene map encoding the bicistronic transcript-regulated promoter expression system of hIL-21 and hIL-15. Figure 3 shows a plastid gene map of the bicistronic transcript-regulated promoter expression system encoding mIL-12. Figure 4 shows a plastid gene map encoding the bicistronic transcript-regulated promoter expression system of mIL-21 and mIL-15. Figure 5 shows the plastid gene map of the regulatory promoter expression system of hIL-12. Figure 6 shows the plastid gene map of the regulated promoter expression system of ml L-12. Figure 7 shows the coding of hIL-12 and hIL-21. The cistron transcript regulates the plastid gene map of the promoter expression system. Figure 8 shows a plastid gene map encoding the tricistronic transcript of mIL-12 and mIL-21 -276-201207108 promoter expression system. Figure 9 shows the structure of the vector rAd.RheoIL12 in which the E1 and E3 regions have been deleted and the RheoSwitch® therapeutic system (RTS)-IL-12 component replaces the E1 region. The box labeled "IL12" represents the IL-12p40 and IL-12p3 5 coding sequences separated by IRES. Figure 10 shows a summary of the transcription, post-transcription, translation and post-translation processing. Figure 11 shows the modular components of a representative production fulcrum. Figure 12 shows the adenoviral compatible ULTRAVECTORtmD backbone with the switch and inducible module synthesis genes. Figure 13 shows an adenoviral vector map of the host-regulated promoter expression system (vector 43 3 18), which contains TNFwt 5MJTR, TNFwtUV signal peptide, TNFwt UV open reading frame (ORF) and SV40e + pA 3' adjustment zone. Figure 14 shows the adenoviral vector map of the regulatory promoter expression system (vector 433 1 9), which contains the TNFwt 5'UTR, the TNFoptUV signal peptide, the TNFopt UV ORF and the 3' of SV40e + pA Adjustment zone. Figure 15 shows the adenoviral vector map of the host-regulated promoter expression system (vector 43320), which regulates the system comprising TNFwt 5'UTR, IL-2optUV signal peptide, TNFopt UV ORF and SV40e + pA3, regulation Area. Figure 16 shows an adenoviral vector map of the ride-regulated promoter expression system (vector 43 3 2 1 ), the regulatory promoter expression system comprising 5U2 5' UTR, -277-201207108 TNFwtUV signal peptide, TNFwt UV ORF and SV40e + 3 of pA, the regulatory zone. Figure 17 shows the adenoviral vector map of the host-regulated promoter expression system (vector 43322), which contains 5U2 5'UTR, TNFoptUV signal peptide, TNFopt UV ORF and SV40e + pA . Figure 18 shows the adenoviral vector map of the host-regulated promoter expression system (vector 43323), which regulates the 5U2 5' UTR, IL-2optUV signal peptide, TNFopt UV ORF and SV40e + pA Area. Figure 19 shows an adenoviral vector gene map of the locus-regulated promoter expression system (vector 43324) 'This regulatory promoter expression system comprises TNFwt 5' UTR, TNFwtUV signal peptide, TNFwt UV ORF and h G Η + p A 3, Adjustment zone. Figure 20 is a diagram showing the adenoviral vector gene map of the host-regulated promoter expression system (vector 43 325), which comprises a TNFwt 5'UTR, a TNFoptUV signal peptide, a TNFopt UV ORF and a hGH + pA 3, regulatory region . Figure 21 shows an adenoviral vector map of the ride-regulated promoter expression system (vector 43 326) containing the TNFwt 5' UTR, IL-2optU V signal peptide, TNFopt UV ORF and hGH + p A 3 'Regulation area. Figure 22 shows an adenoviral vector gene map (vector 43327) of the multiplication-regulated promoter expression system comprising 5U2 5' UTR, -278-201207108 TNFwtUV signal peptide, TNFwt UV ORF and hGH + pA 3 , regulatory region ο Figure 23 shows the adenoviral vector map of the host-regulated promoter expression system (vector 43328), the regulatory promoter expression system comprising 5U2 5'UTR, TNFwtUV signal peptide, TNFwt UV ORF and hGH + pA 3 'Regulatory Region 〇 Figure 24 shows the adenoviral vector map of the host-regulated promoter expression system (vector 43329), which contains 5U2 5' UTR, TNFwtUV signal, TNFwt UV ORF and hGH + pA 'Regulatory Region' Figure 25 shows the adenoviral vector map of the host-regulated promoter expression system (vector 43 5 3 3 ), which contains the TNFwt5'UTR, TNFwtUV signal peptide, TNFwtUV ORF and TNFwt 3'UTR . Figure 26 shows the adenoviral vector map of the ride-regulated promoter expression system (vector 43 5 3 4), which contains the TNFwt5iUTR, TNF full-length ORF and TNFwt 3'UTR. Figure 27 shows the normalized protein secretion of TNF-α induced by RHEOSWITCH® ligand after transfection of HEK293 cells with a vector containing different PT3 components (-/+). Figure 28 shows the normalized protein secretion of TNF-a induced by RHEOSWITCH® ligand after transfection of CHC&gt;_K1 cells with a vector containing different PT3 components (-/+). Figure 29 shows the fold difference in TNF-a secretion by HEK2 93 cells after transfection -279-201207108 Figure 30 shows the difference in protein secretion between 5U2 5'UTR and wtUV TNF-α 5^JTR. Figure 31 is a line graph showing the Ad-RTS-IL-12 dose-dependent assay in the mouse B16F0 melanoma model. Mice were treated with a single injection of Ad-RTS-IL-12 dose lee7, lee8, 1ee9 '5ee9, leelO, 5eel0 virus particles (vp) on day 12, and the ligand was delivered using feed from day 11. The X-axis shows the number of days after tumor cell inoculation, and the Y-axis shows tumor volume. The dose showed a significant anti-tumor effect. The % tumor reduction compared to the control is shown. The line graph of Figure 32 shows the change in body weight during the test. The display is changed by the weight % on the Y axis. Figures 33A and 33B are line graphs showing the anti-tumor activity (Figure 33A) and safety (Figure 33B) of Ad-RTS-mIL12 in the Lewis mouse lung cancer (LLC) model. : Lewis lung tumors were grown subcutaneously in immunocompetent C57b/6 mice. When the tumor grows to the desired size, treatment begins. Animals received a single dose of AdRTS-mIL12 (lelO vp) on days 6, 9, and 13 after tumor cell inoculation. Ad-RTS-mlL 12 showed significant anti-tumor activity compared to control animals compared to control animals. No significant toxicity was observed. The line graphs of Figures 34A and 34B show the efficacy (Figure 34A) and safety (Figure 34B) in a mouse melanoma (B16F0) model in which the animals were treated with Ad-11 D8-11111^12. Tumors developed subcutaneously in the flank of sputum 5715/6 mice. A single dose of human (1-11 D 5-11111^12 (1610乂?) was administered intratumorally on the 13th day after the cell inoculation (arrow). Activated ligand (50, 1〇〇, 25 0, 500, 75 0 and 1 0 0 0 mg/kg) were administered 24 hours before the vehicle injection until the end of the experiment. Tumor growth inhibition was shown as a percentage of the control animals. -280- 201207108

Ad-RTS-IL12 shows therapeutic benefit with a wide range of activated ligand dose windows. The treatment is well tolerated. Figure 35 is a line graph of human and 358 showing the efficacy (Figure 35A) and safety (Figure 35B) of human (1- dent 8-11111^12) in a mouse colon cancer (CT26Luc) model. Mouse colon tumors in Balb The flank of the /C mouse was subcutaneously grown. The animals were treated with the Ad-RTS-mIL12 dose of lelO vp/100 microliters intratumorally (it) on the 11th and 18th day after the cell inoculation (arrow). The activated ligand was The vehicle was administered 24 hours prior to injection. Tumor volume and body weight were monitored throughout the trial. Ad-RTS-mIL12 treatment resulted in superior tumor growth inhibition (100%) relative to control animals. It is noted that all received Ad- The animals treated with RTS-mIL12 plus activating ligands were tumor-free. The line graphs of Figures 36A and 36B show the efficacy (Figure 36A) and safety (Figure 36B) of Ad-RTS-mIL12 in the pancreatic cancer (PAN02) model. PAN02 tumor-bearing mice received a single Ad-RTS-mIL12 dose of lelO vp/100 microliters of tumor (it) on days 7 and 14 after tumor cell inoculation (arrow). Activated ligand feed was administered on vehicle. Animals were supplied from the previous day until the end of the trial. Control or vehicle-only groups were fed only normal mice. Feed. The results showed that Ad-RTS-IL12 exhibited significant anti-tumor activity (97%) relative to control animals. No significant body weight change was found as a result of Ad-RTS-IL12 treatment. Figure 37 is a vector map of AD-RTS-mIFNa Figure 38 is a vector map of AD-RTS-mTNFa. The line graphs of Figures 39A and 39B show the efficacy of Ad-RTS-mIL12 in breast cancer (4T1) model (Figure 39A) and safety (Figure 39B). 4T1 tumor The mice were grown subcutaneously (sc) in the flank of BALB/C mice. Tumor-bearing mice were randomly assigned -281 - 201207108 to 4 groups of 5 animals each; control untreated group, only activated ligand (L) group, Ad-RTS-mIL12 and Ad-RTS-mIL12 plus activation ligand group. Single injection of Ad-RTS-mIL12 was performed at three different time points (arrows) through the tumor (it) at dose lelO ν·ρ·/1 〇〇 was given in microliters of PBS. The activating ligand was supplied to the animals via the feed 24 hours before the vehicle injection until the end of the experiment. Tumor size (volume) and body weight (%) were shown as mean 値±SE. The treated animals (control) had rapid tumor growth. Only activated ligand or three doses of Ad_RTS-miL12 but no activation The treatment had a slight tumor growth inhibition of 22% and 35% compared to the control group. It is noted that the treatment of Ad-RTS-IL 12 plus activating ligand resulted in 82% of the treatment compared to the untreated control animals. Significant tumor growth inhibition. No significant weight loss was observed in any of the treatment groups. Figure 40 shows the vector map of interferon a-2a. -282- 201207108 Sequence Listing &lt;ll0&gt; ^(cuthberts〇n· &lt;120&gt; Vectors for Conditional Expression of Therapeutic Proteins, Host Cells Containing the Vectors, and Their Uses (Vectors Conditionally Expressing Therapeutic Proteins, Host Comprising the Vectors and Uses Thereof) ' &lt;140>TW 100109913 &lt;141&gt; 2011-03-23 &lt;150&gt; US61/431,364 &lt;151&gt; 2011-01-10 &lt;150&gt; US61/366,731 &lt;151&gt; 2010- 07-22 &lt;150&gt; US61/316,792 &lt;151&gt; 2010-03-23 &lt;160> 62

&lt;170&gt; Patent In version 3.5 &lt;210&gt; 1 &lt;211&gt; 13294 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic ιηΙΙ^12 and mIL-21 &lt;400&gt; 1 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 10S0 1140 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc atcatcaata atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggagatccgg taccgatatc ctagacaacg atgctgagct aactataacg gtcctaaggt agcgaccgcg gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata tctttatttt cattacatc t gtgtgttggt tttttgtgtg aatccatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtccaggtg ccagaacatt tctctatcca taatgcaggg gtaccgggtg atgacggtga aaacctccaa ttgcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agagtccccg gggacctaga gggtatataa tgggtgcctt agctggtgtg tgacctcatc ttcctgtacg cccctgcagg ggcgcgccac gcgtcgaaga aggtgagtaa tcttaacatg ctcttttttt ttttttttgc taatcccttt tgtgtgctga tgttaggatg acatttacaa caaatgtttg ttcctgacag gaaaaacctt gctgggtacc ttcgttgccg 1200 201207108 gacacttctt gtcctctact ttggaaaaaa ggaattgaga gccgctagcc caccatgtgc 1260 ccccagaagc tgaccatcag ctggttcgcc atcgtgctgc tggtgagccc cctgatggcc 1320 atgtgggagc tggagaagga cgtgtacgtg gtggaggtgg actggacccc cgacgccccc 1380 ggcgagaccg tgaacctgac ttgcgacacc cccgaggagg acgacatcac ctggaccagc 1440 gaccagagac acggcgtcat cggcagcggc aagaccctga ccatcaccgt gaaggagttc 1500 ctggacgccg gacagtacac ctgtcacaag ggcggcgaga ccctgag cca cagccacctg 1560 ttgctgcaca agaaggagaa cggcatctgg agcaccgaga tcctgaagaa cttcaagaac 1620 aagaccttcc tgaagtgcga ggcccccaac tacagcggca gattcacctg tagctggctg 1680 gtgcagagaa acatggacct gaagttcaac atcaagagca gcagcagcag ccccgacagc 1740 agagccgtga catgcggcat ggccagcctg agcgccgaga aggtgaccct ggaccagaga 1800 gactacgaga agtacagcgt gagctgccag gaggacgtga cctgtcccac cgccgaggag 1860 accctgccca tcgagcttgc cctggaagcc agacagcaga acaagtacga gaactacagc 1920 accagcttct tcatcagaga catcatcaag cccgaccccc ccaagaacct ccagatgaag 1980 cccctgaaga acagccaggt ggaggtgtcc tgggagtacc ccgacagctg gagcaccccc 2040 cacagctact tcagcctgaa gttcttcgtg agaatccaga gaaagaagga gaagatgaag 2100 gagaccgagg agggctgcaa ccagaagggc gctttcctgg tggagaaaac cagcaccgag 2160 gtgcagtgca agggcggcaa cgtgtgtgtg caggcccagg acagatacta caacagcagc 2220 tgctccaagt gggcctgcgt gccctgccgc gtgagaagct gaatcgattg cgcaaagctc 2280 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 2340 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg 2400 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg 2460 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca 2520 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 2580 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 2640 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa 2700 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg 2760 cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg 2820 gacgtggttt tcctttgaaa aacacgatct cttaagtcta gcgccaccat gtgccagagc 2880 agatacctgt tgttcctggc taccctggcc ctgctgaacc acctgagcct ggcccgcgtg 2940 atccccgtga gcggccccgc cagatgcctg agccagagca gaaacctgtt gaaaacaacc 3000 gacgacatgg tgaaaaccgc cagagagaag ctgaagcact acagctgcac cgccgaggac 3060 atcgaccacg aggacatcac cagagaccag accagcaccc tgaaaacctg tctgcccctg 3120 gagctgcaca agaacgagag ctgcctggct accagagaga ccagcagcac caccagaggc 3180 agctgcctgc ccccccagaa aaccagcctg at Gatgaccc tgtgcctggg cagcatctac 3240 gaggacctga agatgtacca gaccgagttc caggccatca acgccgccct gcaaaaccac 3300 aaccaccagc agatcatcct ggacaagggc atgttggtgg ccatcgacga gctgatgcag 3360 agcctgaacc acaacggcga gaccctgaga cagaagcccc ccgtgggcga ggccgacccc 3420

Q 201207108 tacagagtga agatgaagct gtgcatcctg ctgcacgcct tcagcaccag agtggtgacc atcaacagag tgatgggcta cctgagcagc gcctgaatcg aatgcgcact cgagtggtat tacgctcaac ttcagaatct cactaaaaga atagatagtc ttcctttaac tgaaaatttt tccttacaaa cagatatgga cgtcactagc accaccatgg agaggaccct ggtgtgcctg gtggtgatct tcctgggcac cgtggcccac aagagcagcc cccagggacc cgacaggctg ctgatccggc tgagacacct gatcgacatc gtggagcagc tgaagattta cgagaacgac ctggaccccg agctgctgtc cgccccccag gacgtgaagg gccactgcga gcacgccgcc ttcgcctgct tccagaaggc caagctgaag cccagcaacc ccggcaacaa caagaccttc atcatcgacc tggtggccca gctgagaagg aggctgcccg ccaggagggg cggcaagaag cagaagcaca tcgccaagtg ccccagctgc gacagctacg agaagcggac ccccaaggag ttcctggaga ggctgaagtg gctgctgcaa aagatgatcc accagcacct gagctgaatc gcctgcgcag catgctcgcg acctaagtcg gccgctaaag tttacgtagc ggccgcgtcg acgatagctt gatgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggc a agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattctga tttaaatcgg tccgcgtacg gcgtggtagg tccgaacgaa tccatggatt accctgttat ccctatccgg agttaacctc gaggacttcg gaacttctag aaccagaccg ttcagtttaa acgctcttct ccccctcgag ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatat aa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct gggtacgtgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac ggcatgcggg ggggggggsg ggcaattggc caccatgggc cccaagaaga aaaggaaggt ggcccccccc accgacgtga gcctgggcga cgagctgcac ctggacggcg aggacgtggc catggcccac gccgacgccc tggacgactt cgacctggac atgctgggcg acggcgacag ccccggcccc ggcttcaccc cccacgacag 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 201207108 cgccccctac ggcgccctgg acatggccga cttcgagttc gagcagatgt tcaccgacgc 5760 cctgggcatc gacgagtacg gcggccatat ggagatgccc gtggacagga ttctggaggc 5820 cgaactcgcc gtggagcaga aaagcgacca gggcgtggag ggccccggcg gaaccggcgg 5880 cagcggcagc agccccaacg accccgtgac caacatctgc caggccgccg acaagcagct 5 940 gttcaccctg gtggagtggg ccaagaggat tccccacttc agcagcctgc ccctggacga 6000 ccaggtgatc ctgctgaggg ccggatggaa cgagctgctg atcgccagct tcagccacag 6060 gagcatcgac gtgagggacg gcatcctgct ggccaccggc ctgcacgtcc ataggaacag 6120 cgcccacagc gccggagtgg gcgccatctt cgacagggtg ctgaccgagc tggtgagcaa 6180 gatgagggac atgaggatgg acaagaccga gctgggctgc ctgagggcca tcatcctgtt 6240 caaccccgag gtgaggggcc tgaaaagcgc ccaggaggtg gagctgctga gggagaaggt 6300 gtacgccgcc ctggaggagt acaccaggac cacccacccc gacgagcccg gcagattcgc 6360 caagctgctg ctgaggctgc ccagcctgag gagcatcggc ctgaagtgcc tggagcacct 6420 gttcttcttc aggctgatcg gcgacgtgcc catcgacacc ttcctgatgg agatgctgga 6480 gagccccagc gacagctgag ccggcaactc gctgtagtaa ttccagcgag aggcagaggg 6540 agcgagcggg cggcgggcta gggtggagga gcccggcgag cagagctgcg ctgcgggcgt 6600 cctgggaagg gagatccgga gcgaataggg ggcttcgcct ctggcccagc cctcccgctg 6660 atcccccagc cagcggtgcg caaccctagc cgcatccacg aaactttgcc catagcagcg 6720 ggcgggcact ttgcactgga acttacaaca cccgagcaag gacgcgactc tcccgacgcg 6780 gg gaggctat tctgcccatt tggggacact tccccgccgc tgccaggacc cgcttctctg 6840 aaaggctctc cttgcagctg cttagacgct ggattttttt cgggtagtgg aaaaccagca 6900 gcctcccgcg accagatctg ccaccatgaa gctgctgagc agcatcgagc aggcttgcga 6960 catctgcagg ctgaagaagc tgaagigcag caaggagaag cccaagtgcg ccaagtgcct 7020 gaagaacaac tgggagtgca gatacagccc caagaccaag aggagccccc tgaccagggc 7080 ccacctgacc gaggtggaga gcaggctgga gaggctggag cagctgttcc tgctgatctt 7140 ccccagggag gacctggaca tgatcctgaa gatggacagc ctgcaagaca tcaaggccci 7200 gctgaccggc ctgttcgtgc aggacaacgt gaacaaggac gccgtgaccg acaggctggc 7260 cagcgtggag accgacatgc ccctgaccct gaggcagcac aggatcagcg ccaccagcag 7320 cagcgaggag agcagcaaca agggccagag gcagctgacc gtgagccccg agtttcccgg 7380 gatcaggccc gagtgcgtgg tgcccgagac ccagtgcgcc atgaaaagga aggagaagaa 7440 ggcccagaag gagaaggaca agctgcccgt gagcaccacc accgtcgatg accacatgcc 7500 ccccatcatg cagtgcgagc cccccccccc cgaggccgcc aggattcacg aggtcgtgcc 7560 caggttcctg agcgacaagc tgctggtgac caacaggcag aagaacatcc cccagctgac 7620 cgccaacc ag cagttcctga tcgccaggct gatctggtat caggacggct acgagcagcc 7680 cagcgacgag gacctgaaaa ggatcaccca gacctggcag caggccgacg acgagaacga 7740 ggagagcgac acccccttca ggcagatcac cgagatgacc atcctgaccg tgcagctgat 7800 cgtggagttc gccaagggcc tgcccggatt cgccaagatc agccagcccg accagatcac 7860 cctgctgaag gcttgcagca gcgaggtgat gatgctgagg gtggccagga ggtacgacgc 7920 201207108

cgccagcgac agcatcctgt tcgccaacaa ccaggcttac accagggaca actacaggaa 7980 ggctggcatg gccgaggtga tcgaggacct cctgcacttc tgcagatgta tgtacagcat 8040 ggccctggac aacatccact acgccctgct gaccgccgtg gtgatcttca gcgacaggcc 8100 cggcctggag cagccccagc tggtggagga gatccagagg tactacctga acaccctgag 8160 gatctacatc ctgaaccagc tgagcggcag cgccaggagc agcgtgatct acggcaagat 8220 cctgagcatc ctgagcgagc tgaggaccct gggaatgcag aacagcaata tgtgtatcag 8280 cctgaagctg aagaacagga agctgccccc cttcctggag gagatttggg acgtggccga 8340 catgagccac acccagcccc cccccatcct ggagagcccc accaacctgt gaatcgatta 8400 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 8460 tgcttaattt gtgaaatttg tgatgctatt gcttaatttg taaccattat aagctgcaat 8520 aaacaagtta ataaaacatt tgcattcatt ttatgtttca ggttcagggg gagatgtggg 8580 aggtttttta aagcaagtaa aacctctaca aatgtggtat ctagagctct tccaaataga 8640 tctggaaggt gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg 8700 gtaaacatat taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgat c 8760 acttggtgct ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagattgag 8820 gtactgaaat gtgtgggcgt ggcttaaggg tgggaaagaa tatataaggt gggggtctta 8880 tgtagttttg tatctgtttt gcagcagccg ccgccgccat gagcaccaac tcgtttgatg 8940 gaagcattgt gagctcatat ttgacaacgc gcatgccccc atgggccggg gtgcgtcaga 9000 atgtgatggg ctccagcatt gatggtcgcc ccgtcctgcc cgcaaactct actaccttga 9060 cctacgagac cgtgtctgga acgccgttgg agactgcagc ctccgccgcc gcttcagccg 9120 ctgcagccac cgcccgcggg attgtgactg actttgcttt cctgagcccg cttgcaagca 9180 gtgcagcttc ccgttcatcc gcccgcgatg acaagttgac ggctcttttg gcacaattgg 9240 attctttgac ccgggaactt aatgtcgttt ctcagcagct gttggatctg cgccagcagg 9300 tttctgccct gaaggcttcc tcccctccca atgcggttta aaacataaat aaaaaaccag 9360 actctgtttg gatttggatc aagcaagtgt cttgctgtct ttatttaggg gttttgcgcg 9420 cgcggtaggc ccgggaccag cggtctcggt cgttgagggt cctgtgtatt ttttccagga 9480 cgtggtaaag gtgactctgg atgttcagat acatgggcat aagcccgtct ctggggtgga 9540 ggtagcacca ctgcagagct tcatgctgcg gggtggtgtt gtagatgatc c agtcgtagc 9600 aggagcgctg ggcgtggtgc ctaaaaatgt ctttcagtag caagctgatt gccaggggca 9660 ggcccttggt gtaagtgttt acaaagcggt taagctggga tgggtgcata cgtggggata 9720 tgagatgcat cttggactgt atttttaggt tggctatgtt cccagccata tccctccggg 9780 gattcatgtt gtgcagaacc accagcacag tgtatccggt gcacttggga aatttgtcat 9840 gtagcttaga aggaaatgcg tggaagaact tggagacgcc cttgtgacct ccaagatttt 9900 ccatgcattc gtccataatg atggcaatgg gcccacgggc ggcggcctgg gcgaagatat 9960 ttctgggatc actaacgtca tagttgtgtt ccaggatgag atcgtcatag gccattttta 10020 caaagcgcgg gcggagggtg ccagactgcg gtataatggt tccatccggc ccaggggcgt 10080 agttaccctc acagatttgc atttcccacg ctttgagttc agatgggggg atcatgtcta 10140 cctgcggggc gatgaagaaa acggtttccg gggtagggga gatcagctgg gaagaaagca 10200 201207108 ggttcctgag cagctgcgac ttaccgcagc 10320 cggtgggccc gtaaatcaca cctattaccg 10260 ggtgcaactg gtagttaaga gagctgcagc tgccgtcatc cctgagcagg ggggccactt aggcgctcgc 10380 cgcccagcga cgttaagcat gtccctgact cgcatgtttt ccctgaccaa atccgccaga tagcagttct tgcaaggaag caaagttttt caacggtttg agaccgtccg 10440 ccgtaggcat gcttttgagc gtttgaccaa gcagttccag gcggtcccac agctcggtca 10500 cctgctctac ggcatctcga tccagcatat ctcctcgttt cgcgggttgg ggcggctttc 10560 gctgtacggc agtagtcggt gctcgtccag acgggccagg gtcatgtctt tccacgggcg 10620 cagggtcctc gtcagcgtag tctgggtcac ggtgaagggg tgcgctccgg gctgcgcgct 10680 ggccagggtg cgcttgaggc tggtcctgct ggtgctgaag cgctgccggt cttcgccctg 10740 cgcgtcggcc aggtagcatt tgaccatggt gtcatagtcc agcccctccg cggcgtggcc 10800 cttggcgcgc agcttgccct tggaggaggc gccgcacgag gggcagtgca gacttttgag 10860 ggcgtagagc ttgggcgcga gaaataccga ttccggggag taggcatccg cgccgcaggc 10920 cccgcagacg gtctcgcatt ccacgagcca ggtgagctct ggccgttcgg ggtcaaaaac 10980 caggtttccc ccatgctttt tgatgcgttt cttacctctg gtttccatga gccggtgtcc 11040 acgctcggtg acgaaaaggc tgtccgtgtc cccgtataca gacttgagag gcctgtcctc 11100 gaccgatgcc cttgagagcc t tcaacccag tcagctcctt ccggtgggcg cggggcatga 11160 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg 11220 cagcgctct g ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc 11280 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg 11340 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg 11400 gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc 11460 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg 11520 acgaccatca gggacagctt caaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 31580 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 11640 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 11700 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 11760 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 11820 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 11880 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 11940 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 12000 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctct gctga 12060 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 12120 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 12180 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 12240 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 12300 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 12360 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 12420

201207108 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac taattctctt caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaa &lt; 210 &gt; 2 &lt; 211 &gt; 13 333 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220> &lt; 223 &gt; Synthesis of hIL_12 and hIL-21 &lt; 400 &gt; 2 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc atcatcaata atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggagatccgg taccgatatc ctagacaacg atgctgagct aactataacg gtcctaaggt agcgaccgcg gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata tctttatttt cattacatct Gtgtgttggt tttttgtgtg aatccatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtccaggtg ccagaacatt tctctatcca taatgcaggg gtaccgggtg atgacggtga aaacctccaa ttgcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agagtccccg gggacctaga gggtatataa tgggtgcctt agctggtgtg tgacctcatc ttcctgtacg cccctgcagg ggcgcgccac gcgtcgaaga aggtgagtaa 12480 12540 12600 12660 12720 12780 12840 12900 12960 13020 13080 13140 13200 13260 13294 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 3080 201207108 tcttaacatg ctcttttttt ttttttttgc taatcccttt tgtgtgctga tgttaggatg 1140 acatttacaa caaatgtttg ttcctgacag gaaaaacctt gctgggtacc ttcgttgccg 1200 gacacttctt gtcctctact ttggaaaaaa ggaattgaga gccgctagcc caccatgggt 1260 caccagcagt tggtcatctc ttggttttcc ctggtttttc tggcatctcc cctcgtggcc 1320 atatgggaac tgaagaaaga tgtttatgtc gtagaattgg attggtatcc ggatgcccct 1380 ggagaaatgg tggtcctcac ctgtgacacc cctgaagaag Atggtatcac ctggaccttg 1440 gaccagagca gtgaggtctt aggctctggc aaaaccctga ccatccaagt caaagagttt 1500 ggagatgctg gccagtacac ctgtcacaaa ggaggcgagg ttctaagcca ttcgctcctg 1560 ctgcttcaca aaaaggaaga tggaatttgg tccactgata ttttaaagga ccagaaagaa 1620 cccaaaaata agacctttct aagatgcgag gccaagaatt attctggacg tttcacctgc 1680 tggtggctga cgacaatcag tactgatttg acattcagtg tcaaaagcag cagaggctct 1740 tctgaccccc aaggggtgac gtgcggagct gctacactct ctgcagagag agtcagaggg 1800 gacaacaagg agtatgagta ctcagtggag tgccaggagg acagtgcctg cccagctgct 1860 gaggagagtc tgcccattga ggtcatggtg gatgccgttc acaagctcaa gtatgaaaac 1920 tacaccagca gcttcttcat cagggacatc atcaaacctg acccacccaa gaacttgcag 1980 ctgaagccat taaagaattc tcggcaggtg gaggtcagct gggagtaccc tgacacctgg 2040 agtactccac attcciactt ctccctgaca ttctgcgttc aggtccaggg caagagcaag 2100 agagaaaaga aagatagagt cttcacggac aagacctcag ccacggtcat ctgccgcaaa 2160 aatgccagca ttagcgtgcg ggcccaggac cgctactata gctcatcttg gagcgaatgg 2220 gcatctgtgc cctgcagtta gatcgattgc gcaaagctcc ccctctccct cccccccccc 2280 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt 2340 ttccaccata ttgccgtctt ttggcaatgt gagggc ccgg aaacctggcc ctgtcttctt 2400 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt 2460 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct 2520 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt 2580 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt 2640 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa 2700 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta 2760 gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa 2820 acacgatctc ttaagtctag cgccaccatg ggtccagcgc gcagcctcct ccttgtggct 2880 accctggtcc tcctggacca cctcagtttg gccagaaacc tccccgtggc cactccagac 2940 ccaggaatgt tcccatgcct tcaccactcc caaaacctgc tgagggccgt cagcaacatg 3000 ctccagaagg ccagacaaac tctagaattt tacccttgca 3060 cttctgaaga gattgatcat gcctccagaa agacctcttt tatgatggcc gaagatatca caaaagataa aaccagcaca gtggaggcct gtttaccatt ggaattaacc 3120 aagaatgaga gttgcctaaa ttccagagag acctctttca taactaatgg gagttgcctg 3380 ctgtgcctta g Tagtattta tgaagacttg 3240 aagatgtacc aggtggagtt caagaccatg aatgcaaagc ttctgatgga tcctaagagg 3300 201207108

cagatctttc tagatcaaaa catgctggca gttattgatg agctgatgca ggccctgaat 3360 ttcaacagtg agactgtgcc acaaaaatcc tcccttgaag aaccggattt ttataaaact 3420 aaaatcaagc tctgcatact tcttcatgct ttcagaattc gggcagtgac tattgataga 3480 gtgatgagct atctgaatgc ttcctaaatc gaatgcgcac tcgagtggta ttacgctcaa 3540 cttcagaatc tcactaaaag aatagatagt cttcctttaa ctgaaaattt ttccttacaa 3600 acagatatgg acgtcactag caccaccatg agaagcagcc ccggcaacat ggagagaatc 3660 gtgatctgcc tgatggtgat cttcctgggc accctggtgc ataagagcag cagccagggc 3720 caggacagac acatgatccg catgagacag ctgatcgaca tcgtggacca gctgaagaac 3780 tacgtgaacg acctggtgcc cgagttcctg cccgcccccg aggacgtgga gaccaactgc 3840 gagtggagcg ccttcagctg cttccagaag gcccagctga agtccgccaa caccggcaac 3900 aacgagagaa tcatcaacgt gagcatcaag aagctgaagc ggaagccccc cagcaccaac 3960 gccggaagaa gacagaagca cagactgacc tgtcccagct gcgacagcta cgagaagaag 4020 ccccccaagg agttcctgga gagattcaag agcctgctgc aaaagatgat ccaccagcac 4080 ctgagcagca gaacccacgg cagcgaggac agctgaatcg cctgcgcagc atgctcgcg a 4140 cctaagtcgg ccgctaaagt ttacgtagcg gccgcgtcga cgatagcttg atgggtggca 4200 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac 4260 cagccttgtc ctaataaaat taagt tgcat cattttgtct gactaggtgt ccttctataa 4320 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag 4380 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc 4440 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt 4500 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac 4560 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc 4620 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa 4680 ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc 4740 ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta 4800 gatgcctgtt gaattctgat ttaaatcggt ccgcgtacgg cgtggtaggt ccgaacgaat 4860 ccatggatta ccctgttatc cctatccgga gttaacctcg aggacttcgg aacttctaga 4920 accagaccgt tcagtttaaa cgctcttctc cccctcgagg gcctccgcgc cgggttttgg 4980 cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca 5040 gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact 5100 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta 5160 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg 5220 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg 5280 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc 5340 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg ggtacgtgcg ctcggggttg 5400 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata 5460 tgtaattttc agtgttagac tagtaaat tg tccgctaaat tctggccgtt tttggctttt 5520 ttgttagacg gcatgcgggg gggggggggg gcaattggcc accatgggcc ccaagaagaa 5580 201207108 aaggaaggtg gcccccccca ccgacgtgag cctgggcgac gagctgcacc tggacggcga 5640 ggacgtggcc atggcccacg ccgacgccct ggacgacttc gacctggaca tgctgggcga 5700 cggcgacagc cccggccccg gcttcacccc ccacgacagc gccccctacg gcgccctgga 5760 catggccgac ttcgagttcg agcagatgtt cacc gacgcc ctgggcatcg acgagtacgg 5820 cggccatatg gagatgcccg tggacaggat tctggaggcc tggagcagaa gaactcgccg 6180 cgccatcttc gacagggtgc 5880 aagcgaccag ggcgtggagg gccccggcgg aaccggcggc agcggcagca gccccaacga 5940 ccccgtgacc aacatctgcc aggccgccga caagcagctg ttcaccctgg tggagtgggc 6000 caagaggatt ccccacttca gcagcctgcc cctggacgac caggtgatcc tgctgagggc 6060 cggatggaac gagctgctga tcgccagctt cagccacagg agcatcgacg tgagggacgg 6120 catcctgctg gccaccggcc tgcacgtcca taggaacagc gcccacagcg ccggagtggg tgaccgagct ggtgagcaag atgagggaca tgaggatgga 6240 caagaccgag ctgggctgcc tgagggccat catcctgttc aaccccgagg tgaggggcct 6300 gaaaagcgcc caggaggtgg agctgctgag ggagaaggtg tacgccgccc tggaggagta 6360 caccaggacc acccaccccg acgagcccgg cagat tcgcc aagctgctgc tgaggctgcc 6420 cagcctgagg agcatcggcc tgaagtgcct ggagcacctg ttcttcttca ggctgatcgg 6480 cgacgtgccc atcgacacct tcctgatgga gatgctggag agccccagcg acagctgagc 6540 cggcaactcg ctgtagtaat tccagcgaga ggcagaggga gcgagcgggc ggcgggctag 6600 ggtggaggag cccggcgagc agagct gcgc tgcgggcgtc ctgggaaggg agatccggag 6660 cgaatagggg gcttcgcctc tggcccagcc ctcccgctga tcccccagcc agcggtgcgc 6720 aaccctagcc gcatccacga aactttgccc atagcagcgg gcgggcactt tgcactggaa 6780 cttacaacac ccgagcaagg acgcgactct cccgacgcgg ggaggctatt ctgcccattt 6840 ggggacactt ccccgccgct gccaggaccc gcttctctga aaggctctcc ttgcagctgc 6900 ttagacgctg gatttttttc gggtagtgga aaaccagcag cctcccgcga ccagatctgc 6960 caccatgaag ctgctgagca gcatcgagca ggcttgcgac atctgcaggc tgaagaagct 7020 gaagtgcagc aaggagaagc ccaagtgcgc caagtgcctg aagaacaact gggagtgcag 7080 atacagcccc aagaccaaga ggagccccct gaccagggcc cacctgaccg aggtggagag 7140 caggctggag aggctggagc agctgttcct gctgatcttc cccagggagg acctggacat 7200 gatcctgaag atggacagcc tgcaagacat caaggccctg ctgaccggcc tgttcgtgca 7260 ggacaacgtg aacaaggacg ccgtgaccga caggctggcc agcgtggaga ccgacatgcc 7320 cctgaccctg aggcagcaca ggatcagcgc caccagcagc agcgaggaga gcagcaacaa 7380 gggccagagg cagctgaccg tgagccccga gtttcccggg atcaggcccg agtgcgtggt 7440 gcccgagacc cagtgcgcc a tgaaaaggaa ggagaagaag gcccagaagg agaaggacaa 7500 gctgcccgtg agcaccacca ccgtcgatga ccacatgccc cccatcatgc agtgcgagcc 7560 cccccccccc gaggccgcca ggattcacga ggtcgtgccc aggttcctga gcgacaagct 7620 gctggtgacc aacaggcaga agaacatccc ccagctgacc gccaaccagc agttcctgat 7680 cgccaggctg atctggtatc aggacggcta cgagcagccc agcgacgagg acctgaaaag 7740 gatcacccag acctggcagc aggccgacga cgagaacgag gagagcgaca cccccttcag 7800 10 -

C

201207108 gcagatcacc gagatgacca tcctgaccgt gcagctgatc gtggagttcg ccaagggcct gcccggattc gccaagatca gccagcccga ccagatcacc ctgctgaagg cttgcagcag cgaggtgatg atgctgaggg tggccaggag gtacgacgcc gccagcgaca gcatcctgtt cgccaacaac caggcttaca ccagggacaa ctacaggaag gctggcatgg ccgaggtgat cgaggacctc ctgcacttct gcagatgtat gtacagcatg gccctggaca acatccacta cgccctgctg accgccgtgg tgatcttcag cgacaggccc ggcctggagc agccccagct ggtggaggag atccagaggt actacctgaa caccctgagg atctacatcc tgaaccagct gagcggcagc gccaggagca gcgtgatcta cggcaagatc ctgagcatcc tgagcgagct gaggaccctg ggaatgcaga acagcaatat gtgtatcagc ctgaagctga agaacaggaa gctgcccccc ttcctggagg agatttggga cgtggccgac atgagccaca cccagccccc ccccatcctg gagagcccca ccaacctgtg aatcgattag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gcttaatttg tgaaatttgt gatgctattg cttaatttgt aaccattata agctgcaata aacaagttaa taaaacattt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatc tagagctctt ccaaatagat ciggaaggtg ctgaggtacg atgagacccg caccaggtgc agaccctgcg agtgtggcgg taaacatatt aggaaccagc ctgtgatgct ggatgtgacc gaggagctga ggcccgatca cttggtgctg gcctgcaccc gcgctgagtt tggctctagc gatgaagata cagattgagg tactgaaatg tgtgggcgtg gcttaagggt gggaaagaat atataaggtg ggggtcttat gtagttttgt atctgttttg cagcagccgc cgccgccatg agcaccaact cgtttgatgg aagcattgtg agctcatatt tgacaacgcg catgccccca tgggccgggg tgcgtcagaa tgtgatgggc tccagcattg atggtcgccc cgtcctgccc gcaaactcta ctaccttgac ctacgagacc gtgtctggaa cgccgttgga gactgcagcc tccgccgccg cttcagccgc tgcagccacc gcccgcggga ttgtgactga ctttgctttc ctgagcccgc ttgcaagcag tgcagcttcc cgttcatccg cccgcgatga caagttgacg gctcttttgg cacaattgga ttctttgacc cgggaactta atgtcgtttc tcagcagctg ttggatctgc gccagcaggt ttctgccctg aaggcttcct cccctcccaa tgcggtttaa aacataaata aaaaaccaga ctctgtttgg atttggatca agcaagtgtc ttgctgtctt tatttagggg ttttgcgcgc gcggtaggcc cgggaccagc ggtctcggtc gttgagggtc ctgtgtattt tttccaggac gtggtaaagg tgactctgga tgttcagata catgggcata agcccgtctc tggggtggag gtagcaccac tgcagagctt catgctgcgg ggtggtgttg tagatgatcc agtcgtagca ggagcgctgg gcgtggtgcc taaaaatgtc tttcagtagc aagctgattg ccaggggcag gcccttggtg taagtgttta caaagcggtt aagctgggat gggtgcatac gtggggatat gagatgcatc ttggactgta tttttaggtt ggctatgttc ccagccatat ccctccgggg attcatgttg tgcagaacca ccagcacagt gtatccggtg cacttgggaa atttgtcatg tagcttagaa ggaaatgcgt ggaagaactt ggagacgccc ttgtgacctc caagattttc catgcattcg tccataatga tggcaatggg cccacgggcg gcggcctggg cgaagatatt tctgggatca ctaacgtcat agttgtgttc caggatgaga tcgtcatagg ccatttttac aaagcgcggg cggagggtgc -11 - 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 9720 9780 9840 9900 9960 10020 10080 201207108 cagactgcgg tataatggtt ccatccggcc caggggcgta gttaccctca cagatttgca 10140 tttcccacgc tttgagttca gatgggggga tcatgtctac Ctgcgggggg atgaagaaaa 10200 cggtttccgg ggtaggggag atcagctggg aagaaagcag gttcctgagc agctgcgact 10260 taccgcagcc ggtgggcccg taaatcacac ctattaccgg gtgcaactgg tagttaagag 10320 agctgcagct gccgtcatcc ctgagcaggg gggccacttc gttaagcatg tccctgactc 10380 gcatgttttc cctgaccaaa tccgccagaa ggcgctcgcc gcccagcgat agcagttctt 10440 gcaaggaagc aaagtttttc aacggtttga gaccgtccgc cgtaggcatg cttttgagcg 10500 tttgaccaag cagttccagg cggtcccaca gctcggtcac ctgctctacg gcatctcgat 10560 ccagcatatc tcctcgtttc gcgggttggg gcggctttcg ctgtacggca gtagtcggtg 10620 ctcgtccaga cgggccaggg tcatgtcttt ccacgggcgc agggtcctcg tcagcgtagt 10680 ctgggtcacg gtgaaggggt gcgctccggg ctgcgcgctg gccagggtgc gcttgaggct 10740 ggtcctgctg gtgctgaagc gctgccggtc ttcgccctgc gcgtcggcca ggtagcattt 10800 gaccatggtg tcatagtcca gcccctccgc ggcgtggccc ttggcgcgca gcttgccctt 10860 ggaggaggcg ccgcacgagg ggcagtgcag acttttgagg gcgtagagct tgggcgcgag 10920 aaataccgat tccggggagt aggcatccgc gccgcaggcc ccgcagacgg tctcgcattc 10980 cacgagccag gtgagctctg gccgttcggg gtcaaaaacc aggtttcccc catgcttttt 11040 gatgcgtttc ttacctctgg tttccatgag ccggtgtcca cgctcggtga cgaaaaggct 11100 gtccgtgtcc ccgtatacag acttgagagg cctgtcctcg accgatgccc ttgagagcct 11160 tcaacccagt cagctcct tc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga 11220 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg 11280 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa 11340 tct tgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga 11400 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg 11460 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga 11520 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc 11580 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 11640 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 11700 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 11760 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 11820 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 11880 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 11940 agtccaacc c ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 12000 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 12060 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 12120 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttmtgttt 12380 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 12240 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 12300

201207108 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa Atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaa &lt;210&gt; 3 &lt;211&gt; 11553 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; Synthetic mIL-21 and mIL -15 &lt; 400 &gt; 3 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca atcatcaata attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc aagtgacaat tttcgcgcgg ttttaggcgg atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg atgttgtagt aaatttgggc Gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggagatccgg taccgatatc ctagacaacg atgctgagct aactataacg gtcctaaggt agcgaccgcg gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taatgcaggt cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga -13 - 12360 12420 12480 12540 12600 12660 12720 12780 12840 12900 12960 13020 13080 13140 13200 13260 13320 13333 60 120 ISO 240 300 360 420 480 540 600 660 720 780 840 900 201207108 gtactgtcct ccgagcggag tactgtcctc cgagcggaga ctcttcgaag gaagaggggc 960 ggggtcgatc gaccccgccc ctcttcct tc gaaggaagag gggcggggtc gaagacctag 1020 agggtatata atgggtgcct tagctggtgt gtgagctcat cttcctgtag atcacgcgtc 1080 gaagaaggtg agtaatctta acatgctctt tttttttttt tttgctaatc ccttttgtgt 1140 gctgatgtta ggatgacatt tacaacaaat gtttgttcct gacaggaaaa accttgctgg 1200 gtaccttcgt tgccggacac Ttcttgtcct ctactttgga aaaaaggaat tgagagccgc 1260 tagcgccacc atggagagga ccctggtgtg cctggtggtg atcttcctgg gcaccgtggc 1320 ccacaagagc agcccccagg gacccgacag gctgc tgatc cggctgagac acctgatcga 1380 catcgtggag cagctgaaga tttacgagaa cgacctggac cccgagctgc tgtccgcccc 1440 ccaggacgtg aagggccact gcgagcacgc cgccttcgcc tgcttccaga aggccaagct 1500 gaagcccagc aaccccggca acaacaagac cttcatcatc gacctggtgg cccagctgag 1560 aaggaggctg cccgccagga ggggcggcaa gaagcagaag cacatcgcca agtgccccag 1620 ctgcgacagc tacgagaagc ggacccccaa ggagttcctg gagaggctga agtggctgct 1680 gcaaaagatg atccaccagc acctgagctg agttgggcga gctcgaattc attgatcccc 1740 cgggctgcag gaattcgata tcaagctcgg gatccgaati ccgccccccc cccccccccc 1800 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt 1860 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct 1920 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga 1980 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga 2040 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac 2300 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag 2160 ttgtggaaag agtcaaatgg ctctcctc aa gcgtattcaa caaggggctg aaggatgccc 2220 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg 2280 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt 2340 gaaaaacacg atgataatat ggccacaacc atgaagatcc tgaagcccta catgaggaac 2400 accagcatca gctgttacct gtgcttcctg ctgaacagcc acttcctgac cgaggccgga 2460 atccacgtct tcatcctggg ctgcgtgagc gtgggcctgc ccaagaccga ggccaactgg 2520 atcgacgtga ggtacgacct ggagaagatc gagagcctga tccagagcat ccacatcgac 2580 accaccctgt acaccgacag cgacttccac cccagctgca aggtgaccgc catgaactgc 2640 ttcctgctgg agctgcaagt gatcctgcac gagtacagca acatgaccct gaacgagacc 2700 gtgaggaacg tgctgtacct ggctaacagc accctgagca gcaacaagaa cgtggccgag 2760 agcggctgca aggagtgtga ggagctggag gagaagacct tcaccgagtt cctccagagc 2820 ttcatcagga tcgtgcagat gttcatcaac accagctgaa tcgattgcgc aaagctt teg 2880 cgataggcga gaccaatggg tgtgtacgta gcggccgctc gagaacttgt ttattgeage 2940 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 3000 actgcattct agttgtggtt Tgtccaaact catcaatgta tcttatcatg tetegtaegg 3060 cgtggtaggt ccgaacgaat ccatggatta ccctgttatc cctatccgga gttaacctcg 3120 -14 -

C 201207108 aggacttcgg aacttctaga accagaccgt tcagtttaaa cgctcttctc cccctcgagg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg ggtacgtgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagacg gcatgcgggg gggggggggg gcaattggcc accatgggcc ccaagaagaa aaggaaggtg gcccccccca ccgacgtgag cctgggcgac gagctgcacc tggacggcga ggacgtggcc atggcccacg ccgacgccct ggacgacttc gacctggaca tgctgggcga cggcgacagc cccggccccg gcttcacccc ccacgacagc gccccctacg gcgccctgga catggccgac ttcgagttcg agcagatgtt caccgacgc c ctgggcatcg acgagtacgg cggccatatg gagatgcccg tggacaggat tctggaggcc gaactcgccg tggagcagaa aagcgaccag ggcgtggagg gccccggcgg aaccggcggc agcggcagca gccccaacga ccccgtgacc aacatctgcc aggccgccga caagcagctg ttcaccctgg tggagtgggc caagaggatt ccccacttca gcagcctgcc cctggacgac caggtgatcc tgctgagggc cggatggaac gagctgctga tcgccagctt cagccacagg agcatcgacg tgagggacgg catcctgctg gccaccggcc tgcacgtcca taggaacagc gcccacagcg ccggagtggg cgccatcttc gacagggtgc igaccgagct ggtgagcaag atgagggaca tgaggatgga caagaccgag ctgggctgcc tgagggccat catcctgttc aaccccgagg tgaggggcct gaaaagcgcc caggaggtgg agctgctgag ggagaaggtg tacgccgccc tggaggagta caccaggacc acccaccccg acgagcccgg cagattcgcc aagctgctgc tgaggctgcc cagcctgagg agcatcggcc tgaagtgcct ggagcacctg ttcttcttca ggctgatcgg cgacgtgccc atcgacacct tcctgatgga gatgctggag agccccagcg acagctgagc cggcaactcg ctgtagtaat tccagcgaga ggcagaggga gcgagcgggc ggcgggctag ggtggaggag cccggcgagc agagctgcgc tgcgggcgtc ctgggaaggg agatccggag cgaatagggg gcttcgcctc tggcccagcc ctcccgctga tcccccag cc agcggtgcgc aaccctagcc gcatccacga aactttgccc atagcagcgg gcgggcactt tgcactggaa cttacaacac ccgagcaagg acgcgactct cccgacgcgg ggaggctatt ctgcccattt ggggacactt ccccgccgct gccaggaccc gcttctctga aaggctctcc ttgcagctgc ttagacgctg gatttttttc gggtagtgga aaaccagcag cctcccgcga ccagatctgc caccatgaag ctgctgagca gcatcgagca ggcttgcgac atctgcaggc tgaagaagct gaagtgcagc aaggagaagc ccaagtgcgc caagtgcctg aagaacaact gggagtgcag atacagcccc aagaccaaga ggagccccct gaccagggcc cacctgaccg aggtggagag caggctggag aggctggagc agctgttcct gctgatcttc -15 - 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 201207108 cccagggagg acctggacat gatcctgaag atggacagcc tgcaagacat caaggccctg 5460 ctgaccggcc tgttcgtgca ggacaacgtg Aacaaggacg ccgtgaccga caggctggcc 5520 agcgtggaga ccgacatgcc cctgaccctg aggcagcaca ggatcagcgc caccagcagc 5580 agcgaggaga gcagcaacaa gggccagagg cagctgaccg tgagccccga gtt tcccggg 5640 atcaggcccg agtgcgtggt gcccgagacc cagtgcgcca tgaaaaggaa ggagaagaag 5700 gcccagaagg agaaggacaa gctgcccgtg agcaccacca ccgtcgatga ccacatgccc 5760 cccatcatgc agtgcgagcc cccccccccc gaggccgcca ggattcacga ggtcgtgccc 5820 aggttcctga gcgacaagct gciggtgacc aacaggcaga agaacatccc ccagctgacc 5880 gccaaccagc agttcctgat cgccaggctg atctggtatc aggacggcta cgagcagccc 5940 agcgacgagg acctgaaaag gatcacccag acctggcagc aggccgacga cgagaacgag 6000 gagagcgaca cccccttcag gcagatcacc gagatgacca tcctgaccgt gcagctgatc 6060 gtggagttcg ccaagggcct gcccggattc gccaagatca gccagcccga ccagatcacc 6120 ctgctgaagg cttgcagcag cgaggtgatg atgctgaggg tggccaggag gtacgacgcc 6180 gccagcgaca gcatcctgtt cgccaacaac caggcttaca ccagggacaa ctacaggaag 6240 gctggcatgg ccgaggtgat cgaggacctc ctgcacttct gcagatgtat gtacagcatg 6300 gccctggaca acatccacta cgccctgctg accgccgtgg tgatcttcag cgacaggccc 6360 ggcctggagc agccccagct ggtggaggag atccagaggt actacctgaa caccctgagg 6420 atctacatcc tgaaccagct gagcggcagc gccaggagca gcgtga tcta cggcaagatc 6480 ctgagcatcc tgagcgagct acagcaatat gaggaccctg ggaatgcaga ggttttttaa agcaagtaaa acctctacaa gtgtatcagc 6540 ctgaagctga agaacaggaa gctgcccccc ttcctggagg agatttggga cgtggccgac 6600 atgagccaca cccagccccc ccccatcctg gagagcccca ccaacctgtg aatcgattag 6660 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat 6720 gcttaatttg tgaaatttgt gatgctattg cttaatttgt aaccattata agctgcaata 6780 aacaagttaa taaaacattt gcattcattt tatgtttcag gttcaggggg agatgtggga 6840 atgtggtatc tagagctctt ccaaatagat 6900 ctggaaggtg ctgaggtacg atgagacccg caccaggtgc agaccctgcg agtgtggcgg 6960 taaacatatt aggaaccagc ctgtgatgct ggatgtgacc gaggagctga ggcccgatca 7020 cttggtgctg gcctgcaccc gcgctgagtt tggctctagc gatgaagata cagattgagg 7080 tactgaaatg tgtgggcgtg gcttaagggt gggaaagaat atataaggtg ggggtcttat 7140 gtagttttgt atctgttttg cagcagccgc cgccgccatg agcaccaact cgtttgatgg 7200 aagcattgtg agctcatatt tgacaacgcg catgccccca tgggccgggg tgcgtcagaa 7260 tgtgatgggc tccagcattg atggtcgccc cgtcctgcc c gcaaactcta ctaccttgac 7320 ctacgagacc gtgtctggaa cgccgttgga gactgcagcc tccgccgccg cttcagccgc 7380 tgcagccacc gcccgcggga ttgtgactga ctttgctttc ctgagcccgc ttgcaagcag 7440 tgcagcttcc cgttcatccg cccgcgatga caagttgacg gctcttttgg cacaattgga 7500 ttctttgacc cgggaactta atgtcgtttc tcagcagctg ttggatctgc gccagcaggt 7560 ttctgccctg aaggcttcct cccctcccaa tgcggtttaa aacataaata aaaaaccaga 7620 -16 - f

201207108 ctctgtttgg atttggatca agcaagtgtc ttgctgtctt tatttagggg ttttgcgcgc gcggtaggcc cgggaccagc ggtctcggtc gttgagggtc ctgtgtattt tttccaggac gtggtaaagg tgactctgga tgttcagata catgggcata agcccgtctc tggggtggag gtagcaccac tgcagagctt catgctgcgg ggtggtgttg tagatgatcc agtcgtagca ggagcgctgg gcgtggtgcc taaaaatgtc tttcagtagc aagctgattg ccaggggcag gcccttggtg taagtgttta caaagcggtt aagctgggat gggtgcatac gtggggatat gagatgcatc ttggactgta tttttaggtt ggctatgttc ccagccatat ccctccgggg attcatgttg tgcagaacca ccagcacagt gtatccggtg cacttgggaa atttgtcatg tagcttagaa ggaaatgcgt ggaagaactt ggagacgccc ttgtgacctc caagattttc catgcattcg tccataatga tggcaatggg cccacgggcg gcggcctggg cgaagatatt tctgggatca ctaacgtcat agttgtgttc caggatgaga tcgtcatagg ccatttttac aaagcgcggg cggagggtgc cagactgcgg tataatggtt ccatccggcc caggggcgta gttaccctca cagatttgca tttcccacgc tttgagttca gatgggggga tcatgtctac ctgcggggcg atgaagaaaa cggtttccgg ggtaggggag atcagctggg aagaaagcag gttcctgagc agctgcgact taccgcagcc ggtgggcccg taaatcacac ctattaccgg gtgcaactgg tagttaagag agctgcagct gccgtcatcc ctgagcaggg gggccacttc gttaagcatg tccctgactc gcatgttttc cctgaccaaa tccgccagaa ggcgctcgcc gcccagcgat agcagttctt gcaaggaagc aaagtttttc aacggtttga gaccgtccgc cgtaggcatg cttttgagcg tttgaccaag cagttccagg cggtcccaca gctcggtcac ctgctctacg gcatctcgat ccagcatatc tcctcgtttc gcgggttggg gcggctttcg ctgtacggca gtagtcggtg ctcgtccaga cgggccaggg tcatgtcttt ccacgggcgc agggtcctcg tcagcgtagt ctgggtcacg gtgaaggggt gcgctccggg ctgcgcgctg gccagggtgc gcttgaggct ggtcctgctg gtgctgaagc gctgccggtc ttcgccctgc gcgtcggcca ggtagcattt gaccatggtg tcatagtcca gcccctccgc ggcgtggccc ttggcgcgca gcttgccctt ggaggaggcg ccgcacgagg ggcagtgcag acttttgagg gcgtagagct tgggcgcgag aaataccgat tccggggagt aggcatccgc gccgcaggcc ccgcagacgg tctcgcattc cacgagccag gtgagctctg gccgttcggg gtcaaaaacc aggtttcccc catgcttttt gatgcgtttc ttacctctgg tttccatgag ccggtgtcca cgctcggtga cgaaaaggct gtccgtgtcc ccgtatacag acttgagagg cctgtcctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag -17 ctgtcttctt - 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 9720 9780 9840 9900 201207108 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 9960 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga Tacctgtccg cctttctccc 10020 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 10080 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcg cctt 10140 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 10200 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 10260 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 10320 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 10380 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 10440 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 10500 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 10560 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 10620 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 10680 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 10740 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 10800 aagggccgag cgcagaagtg gtcctgcaac 11tatccgcc tccatccagt ctattaattg 10860 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 10920 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattc a gctccggttc 10980 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 11040 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 11100 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 11160 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 11220 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 11280 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 11340 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 31400 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 11460 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 11520 gagcggatac atatttgaat gtatttagaa aaa 11553 &lt; 210 &gt; 4 &lt; 211 &gt; 12279 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; Synthetic &lt;400&gt; 4 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 60 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaatcgca ccggtatcta tgtcgggtg c ggagaaagag gtaatgaaat ggcagctagc 180 atcatcaata atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt 240

201207108 tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggagatccgg taccgatatc ctagacaacg atgctgagct aactataacg gtcctaaggt agcgaccgcg gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taatgcaggt cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggaga ctcttcgaag gaagaggggc ggggtcgatc gaccccgccc ctcttccttc gaaggaagag gggcggggtc gaagacctag agggtatata atgggtgcct tagctggtgt gtgagctcat cttcctgtag atcacgcgtc gaagaaggtg agtaatctta acatgctctt tttttttttt tttgctaatc ccttttgtgt gctgatgtta ggatgacatt tacaacaaat gtttgttcct gacaggaaaa accttgctgg gtaccttcgt tgccggacac ccgagatcct gaagaacttc aagaacaaga ttcttgtcct ctactttgga aaaaaggaat tgagagccgc tagcgccacc atgtgccccc agaagctgac catcagctgg ttcgccatcg tgctgctggt gagccccctg atggccatgt gggagctgga gaaggacgtg tacgtggtgg aggtggactg gacccccgac gcccccggcg agaccgtgaa cctgacttgc gacacccccg aggaggacga catcacctgg accagcgacc agagacacgg cgtcatcggc agcggcaaga ccctgaccat caccgtgaag gagttcctgg acgccggaca gtacacctgt cacaagggcg gcgagaccct gagccacagc cacctgttgc tgcacaagaa ggagaacggc atctggagca ccttcctgaa gtgcgaggcc cccaactaca gcggcagatt cacctgtagc tggctggtgc agagaaacat ggacctgaag ttcaacatca agagcagcag cagcagcccc gacagcagag ccgtgacatg cggcatggcc agcctgagcg ccgagaaggt gaccctggac cagagagact acgagaagta cagcgtgagc tgccaggagg acgtgacctg tcccaccgcc gaggagaccc tgcccatcga gcttgccctg gaagccagac agcagaacaa gtacgagaac tacagcacca gcttcttcat cagagacatc atcaagcccg acccccccaa gaacctccag atgaagcccc tgaagaacag ccaggtggag gtgtcctggg agtaccccga cagctggagc accccccaca gctacttcag cctgaagttc ttcgtgagaa tccagagaaa gaaggagaag atgaaggaga ccgaggaggg ctgcaaccag aagggcgctt tcctggtgga gaaaaccagc accgaggtgc agtgcaaggg cggcaacgtg tgtgtgcagg cccaggacag atactacaac agcagctgct ccaagtgggc ctgcgtgccc tgccgcgtga gaagctgagt tgggcgagct cgaattcatt gatcccccgg gctgcaggaa ttcgatatca agctcgggat ccgaattccg cccccccccc cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc -19 - 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 I860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 201207108 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 2580 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca Gcggaacccc ccacctggcg 2640 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 2700 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 2760 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 2820 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc 2880 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg 2940 tgccagagca gatacctgtt gttcctggct accctggccc tgctgaacca cctgagcctg 3000 gcccgcgtga tccccgtgag cggccccgcc agatgcctga gccagagcag aaacctgttg 3060 aaaacaaccg acgacatggt gaaaaccgcc agagagaagc tgaagcacta cagctgcacc 3120 gccgaggaca tcgaccacga ggacatcacc agagaccaga ccagcaccct gaaaacctgt 3180 ctgcccctgg agctgcacaa gaacgagagc tgcctggcta ccagagagac cagcagcacc 3240 accagaggca gctgcctgcc cccccagaaa accagcctga tgatgaccct gtgcctgggc 3300 agcatctacg aggacctgaa gatgtaccag accgagttcc aggccatcaa cgccgccctg 3360 caaaaccaca accaccagca gatcatcctg gacaagggca tgttggtggc catcgacgag 3420 ctgatgcaga gcctgaacca caacggcgag accctgagac agaagccccc cgtgggcgag 3480 gccgacccct acagagtgaa gatgaagctg tgcatcctgc tgcacgcctt cagcaccaga 3540 gtggtgacca tcaacagagt gatgggctac ctgagcagcg cctgaatcga ttgcgcaaa g 3600 ctttcgcgat aggcgagacc aatgggtgtg tacgtagcgg ccgctcgaga acttgtttat 3660 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 3720 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctc 3780 gtacggcgtg gtaggtccga acgaatccat ggattaccct gttatcccta tccggagtta 3840 acctcgagga cttcggaact tctagaacca gaccgttcag tttaaacgct ct tctccccc 3900 tcgagggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag 3960 cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag 4020 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat 4080 tttaggacgg gacttgggtg actctagggc actggttttc 11 tccagaga gcggaacagg 4140 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg 4200 ccgatgatta 4260 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt ttggtgagta gcgggctgct 4320 gggctgggta cgtgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt 4380 ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt aaattgtccg 4440 ctaaattctg gccgtttttg gcttttttgt tagacggcat gcgggggggg gggggggcaa 4500 ttggccacca tgggccccaa gaagaaaagg aaggtggccc cccccaccga cgtgagcctg 4560 ggcgacgagc tgcacctgga cggcgaggac gtggccatgg cccacgccga cgccctggac 4620 gacttcgacc tggacatgct gggcgacggc gacagccccg gccccggctt caccccccac 4680 gacagcgccc cctacggcgc cctggacatg gccgacttcg agt tcgagca gatgttcacc 4740 -20 -

C

201207108 gacgccctgg gcatcgacga gtacggcggc catatggaga tgcccgtgga caggattctg gaggccgaac tcgccgtgga gcagaaaagc gaccagggcg tggagggccc cggcggaacc ggcggcagcg gcagcagccc caacgacccc gtgaccaaca tctgccaggc cgccgacaag cagctgttca ccctggtgga gtgggccaag aggattcccc acttcagcag cctgcccctg gacgaccagg tgatcctgct gagggccgga tggaacgagc tgctgatcgc cagcttcagc cacaggagca tcgacgtgag ggacggcatc ctgctggcca ccggcctgca cgtccatagg aacagcgccc acagcgccgg agtgggcgcc atcttcgaca gggtgctgac cgagctggtg agcaagatga gggacatgag gatggacaag accgagctgg gctgcctgag ggccatcatc ctgttcaacc ccgaggtgag gggcctgaaa agcgcccagg aggtggagct gctgagggag aaggtgtacg ccgccctgga ggagtacacc aggaccaccc accccgacga gcccggcaga ttcgccaagc tgctgctgag gctgcccagc ctgaggagca tcggcctgaa gtgcctggag cacctgttct tcttcaggct gatcggcgac gtgcccatcg acaccttcct gatggagatg ctggagagcc ccagcgacag ctgagccggc aactcgctgt agtaattcca gcgagaggca gagggagcga gcgggcggcg ggctagggtg gaggagcccg gcgagcagag ctgcgctgcg ggcgtcctgg gaagggagat ccggagcgaa tagggggctt cgcctctggc ccagccctcc cgctgatccc ccagccagcg gtgcgcaacc ctagccgcat ccacgaaact ttgcccatag cagcgggcgg gcactttgca ctggaactta caacacccga gcaaggacgc gactctcccg acgcggggag gctattctgc ccatttgggg acacttcccc gccgctgcca ggacccgctt ctctgaaagg ctctccttgc agctgcttag acgctggatt tttttcgggt agtggaaaac cagcagcctc ccgcgaccag atctgccacc atgaagctgc tgagcagcat cgagcaggct tgcgacatct gcaggctgaa gaagctgaag tgcagcaagg agaagcccaa gtgcgccaag tgcctgaaga acaactggga gtgcagatac agccccaaga ccaagaggag ccccctgacc agggcccacc tgaccgaggt ggagagcagg ctggagaggc tggagcagct gttcctgctg atcttcccca gggaggacct ggacatgatc ctgaagatgg acagcctgca agacatcaag gccctgctga ccggcctgtt cgtgcaggac aacgtgaaca aggacgccgt gaccgacagg ctggccagcg tggagaccga catgcccctg accctgaggc agcacaggat cagcgccacc agcagcagcg aggagagcag caacaagggc cagaggcagc tgaccgtgag ccccgagttt cccgggatca ggcccgagtg cgtggtgccc gagacccagt gcgccatgaa aaggaaggag aagaaggccc agaaggagaa ggacaagctg cccgtgagca ccaccaccgt cgatgaccac atgcccccca tcatgcagtg cgagcccccc ccccccgagg ccgccaggat tcacgaggtc gtgcccaggt tcctgagcga caagctgctg gtgaccaaca ggcagaagaa catcccccag ctgaccgcca accagcagtt cctgatcgcc aggctgatct ggtatcagga cggctacgag cagcccagcg acgaggacct gaaaaggatc acccagacct ggcagcaggc cgacgacgag aacgaggaga gcgacacccc cttcaggcag atcaccgaga tgaccatcct gaccgtgcag ctgatcgtgg agttcgccaa gggcctgccc ggattcgcca agatcagcca gcccgaccag atcaccctgc tgaaggcttg cagcagcgag gtgatgatgc tgagggtggc caggaggtac gacgccgcca gcgacagcat cctgttcgcc aacaaccagg cttacaccag ggacaactac aggaaggctg gcatggccga ggtgatcgag gacctcctgc acttctgcag atgtatgtac -21 - 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 201207108 agcatggccc tggacaacat ccactacgcc ctgctgaccg ccgtggtgat cttcagcgac 7080 aggcccggcc tggagcagcc ccagctggtg gaggagatcc Agaggtacta cctgaacacc 7140 ctgaggatct acatcctgaa ccagctgagc ggcagcgcca ggagcagcgt gatctacggc 7200 aagatcctga gcatcctgag cgagctgagg accctgggaa tgcagaacag caata tgtgt 7260 atcagcctga agctgaagaa caggaagctg ccccccttcc tggaggagat ttgggacgtg 7320 gccgacatga gccacaccca gccccccccc atcctggaga gccccaccaa cctgtgaatc 7380 gattagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa 7440 aaaaatgctt aatitgtgaa atttgtgatg ctattgctta atttgtaacc attataagct 7500 gcaataaaca agttaataaa acatttgcat tcattttatg tttcaggttc agggggagat 7560 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatctaga gctcttccaa 7620 atagatctgg aaggtgctga ggtacgatga gacccgcacc aggtgcagac cctgcgagtg 7680 tggcggtaaa catattagga accagcctgt gatgctggat gtgaccgagg agctgaggcc 7740 cgatcacttg gtgctggcct gcacccgcgc tgagtttggc tctagcgatg aagatacaga 7800 ttgaggtact gaaatgtgtg ggcgtggctt aagggtggga aagaatatat aaggtggggg 7860 tcttatgtag ttttgtatct gttttgcagc agccgccgcc gccatgagca ccaactcgtt 7920 tgatggaagc attgtgagct catat ttgac aacgcgcatg cccccatggg ccggggtgcg 7980 tcagaatgtg atgggctcca gcattgatgg tcgccccgtc ctgcccgcaa actctactac 8040 cttgacctac gagaccgtgt ctggaacgcc gttggagact gcagcct ccg ccgccgcttc 8100 agccgctgca gccaccgccc gcgggattgt gactgacttt gctttcctga gcccgcttgc 8160 aagcagtgca gcttcccgtt catccgcccg cgatgacaag ttgacggctc ttttggcaca 8220 attggattct ttgacccggg aacttaatgt cgtttctcag cagctgttgg atctgcgcca 8280 gcaggtttct gccctgaagg cttcctcccc tcccaatgcg gtttaaaaca taaataaaaa 8340 accagactct gtttggattt ggatcaagca agtgtcttgc tgtctttatt taggggtttt 8400 gcgcgcgcgg taggcccggg accagcggtc tcggtcgttg agggtcctgt gtattttttc 8460 caggacgtgg taaaggtgac tctggatgtt cagatacatg ggcataagcc cgtctctggg 8520 gtggaggtag caccactgca gagcttcatg ctgcggggtg gtgttgtaga tgatccagtc 8580 gtagcaggag cgctgggcgt ggtgcctaaa aatgtctttc agtagcaagc tgattgccag 8640 gggcaggccc ttggtgtaag tgtttacaaa gcggttaagc tgggatgggt gcatacgtgg 8700 ggatatgaga tgcatcttgg actgtatttt taggttggct atgttcccag ccatatccct 8760 ccggggattc atgttgtgca gaaccaccag cacagtgtat ccggtgcact tgggaaattt 8820 gtcatgtagc ttagaaggaa atgcgtggaa gaacttggag acgcccttgt gacctccaag 8880 attttccatg cattcgtcca taatgatggc aatgggccca cgggcggcgg cctgggcgaa 8940 gatatttctg ggatcactaa cgtcatagtt gtgttccagg atgagatcgt cataggccat 9000 ttt tacaaag cgcgggcgga gggtgccaga ctgcggtata atggttccat ccggcccagg 9060 ggcgtagtta ccctcacaga tttgcatttc ccacgctttg agttcagatg gggggatcat 9120 gtctacctgc ggggcgatga agaaaacggt ttccggggta ggggagatca gctgggaaga 9180 aagcaggttc ctgagcagct gcgacttacc gcagccggtg ggcccgtaaa tcacacctat 9240 -22 -

201207108 taccgggtgc aactggtagt taagagagct gcagctgccg tcatccctga gcaggggggc cacttcgtta agcatgtccc tgactcgcat gttttccctg accaaatccg ccagaaggcg ctcgccgccc agcgatagca gttcttgcaa ggaagcaaag tttttcaacg gtttgagacc gtccgccgta ggcatgcttt tgagcgtttg accaagcagt tccaggcggt cccacagctc ggtcacctgc tctacggcat ctcgatccag catatctcct cgtttcgcgg gttggggcgg ctttcgctgt acggcagtag tcggtgctcg tccagacggg ccagggtcat gtctttccac gggcgcaggg tcctcgtcag cgtagtctgg gtcacggtga aggggtgcgc tccgggctgc gcgctggcca gggtgcgctt gaggctggtc ctgctggtgc tgaagcgctg ccggtcttcg ccctgcgcgt cggccaggta gcatttgacc atggtgtcat agtccagccc ctccgcggcg tggcccttgg cgcgcagctt gcccttggag gaggcgccgc acgaggggca gtgcagactt ttgagggcgt agagcttggg cgcgagaaat accgattccg gggagtaggc atccgcgccg caggccccgc agacggtctc gcattccacg agccaggtga gctctggccg ttcggggtca aaaaccaggt ttcccccatg ctttttgatg cgtttcttac ctctggtttc catgagccgg tgtccacgct cggtgacgaa aaggctgtcc gtgtccccgt atacagactt gagaggcctg tcctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc -23 - 9300 9360 9420 9480 9540 9600 9660 9720 9780 9840 9900 9960 10020 10080 10140 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 11160 11220 11280 11340 11400 11460 11520 201207108 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 11580 taattgttgc cgggaagcta gagtaagtag ttcgccagtt Aatagtttgc gcaacgttgt 11640 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 11700 cggttcccaa cgatcaaggc gagttacat g atcccccatg ttgtgcaaaa aagcggttag 11760 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 11820 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 11880 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 11940 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 12000 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 12060 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 12120 tgggtgagca aaaacaggaa ggcaaaatgc Cgcaaaaaag ggaataaggg cgacacggaa 12180 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 12240 tctcatgagc ggatacatat ttgaatgtat ttagaaaaa 12279 &lt;210&gt; 5 &lt;211&gt; 11601 &lt;212&gt; DNA &lt;2]3&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic hIL -21 and hIL-15 &lt;400&gt; 5 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 60 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc 180 atcatcaata atatacct ta ttttggattg aagccaatat gataatgagg gggtggagtt 240 tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg 300 atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg 360 tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt 420 aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa 480 gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggagatccgg 540 taccgatatc ctagacaacg atgctgagct aactataacg gtcctaaggt agcgaccgcg 600 gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata 660 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac 720 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca 780 agtgcaggtg ccagaacatt tctctatcga taatgcaggt cggagtactg tcctccgagc 840 ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga 900 gtactgtcct ccgagcggag tactgtcctc cgagcggaga ctcttcgaag gaagaggggc 960 ggggtcgatc gaccccgccc ctcttccttc gaag Gaagag gggcggggtc gaagacctag 1020 agggtatata atgggtgcct tagctggtgt gtgagctcat cttcctgtag atcacgcgtc 1080 gaagaaggtg agtaatctta acatgctctt tttttttttt tttgctaatc ccttttgtgt 1140 -24 - 201207108

gctgatgtta ggatgacatt tacaacaaat gtttgttcct gacaggaaaa accttgctgg 1200 gtaccttcgt tgccggacac ttcttgtcct ctactttgga aaaaaggaat tgagagccgc 1260 tagcgccacc atgagaagca gccccggcaa catggagaga atcgtgatct gcctgatggt 1320 gatcttcctg ggcaccctgg tgcataagag cagcagccag ggccaggaca gacacatgat 1380 ccgcatgaga cagctgatcg acatcgtgga ccagctgaag aactacgtga acgacctggt 1440 gcccgagttc ctgcccgccc ccgaggacgt ggagaccaac tgcgagtgga gcgccttcag 1500 ctgcttccag aaggcccagc tgaagtccgc caacaccggc aacaacgaga gaatcatcaa 1560 cgtgagcatc aagaagctga agcggaagcc ccccagcacc aacgccggaa gaagacagaa 1620 gcacagactg acctgtccca gctgcgacag ctacgagaag aagcccccca aggagttcct 1680 ggagagattc aagagcctgc tgcaaaagat gatccaccag cacctgagca gcagaaccca 1740 cggcagcgag gacagctgag ttgggcgagc tcgaattcat tgatcccccg ggctgcagga 1800 attcgatatc aagctcggga tccgaattcc gccccccccc cccccccccc cctaacgtta 1860 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca 1920 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagc a 1980 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg 2040 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc 2100 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata 2160 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag 2220 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc 2280 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt 2340 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat 2400 gataatatgg ccacaaccat gagaatcagc aagccccacc tgagaagcat cagcatccag 2460 tgttacctgt gcctgctgct gaacagccac ttcctgaccg aggccggtat ccacgtcttc 2520 atcctgggct gcttcagcgc cggactgccc aagaccgagg ccaactgggt gaacgtgatc 2580 tctgacctga agaagatcga ggacctgatc cagtccatgc acatcgacgc caccctgtac 2640 accgagagcg acgttcatcc cagctgcaag gtgaccgcca tgaagtgctt cctgctggag 2700 ctgcaagtga tctccctgga gagcggcgac gccagcatcc acgacaccgt ggagaacctg 2760 attatcctgg ctaacaacag cctgagcagc aacggcaacg tgaccgagag c ggctgcaag 2820 gagtgtgagg agctggagga gaagaacatc aaggagttcc tccagagctt cgtgcatatc 2880 gtccagatgt tcatcaacac cagctgaatc gattgcgcaa agctttcgcg ataggcgaga 2940 ccaatgggtg tgtacgtagc ggccgctcga gaacttgttt attgcagctt ataatggtta 3000 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 3060 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tcgtacggcg tggtaggtcc 3120 gaacgaatcc atggattacc ctgttatccc tatccggagt taacctcgag gacttcggaa 3180 cttctagaac cagaccgttc agtttaaacg ctcttctccc cctcgagggc ctccgcgccg 3240 ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca cgtcagacga 3300 agggcgcagc gagcgtcctg atccttccgc ccggacgcic aggacagcgg cccgctgctc 3360 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac -25 - gggacttggg 3420 201207108 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct 3480 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga 3540 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt 3600 tgtggatcgc tgtgatcgtc acttggtgagtagcgggctg ctgggctggg tacgtgcgct 3660 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg 3720 ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc tggccgtttt 3780 tggctttttt gttagacggc atgcgggggg gggggggggc aattggccac catgggcccc 3840 aagaagaaaa ggaaggtggc cccccccacc gacgtgagcc tgggcgacga gctgcacctg 3900 gacggcgagg acgtggccat ggcccacgcc gacgccctgg acgacttcga cctggacatg 3960 ctgggcgacg gcgacagccc cggccccggc ttcacccccc acgacagcgc cccctacggc 4020 gccctggaca tggccgactt cgagttcgag cagatgttca ccgacgccct gggcatcgac 4080 gagtacggcg gccatatgga gatgcccgtg gacaggattc tggaggccga actcgccgtg 4140 gagcagaaaa gcgaccaggg cgtggagggc cccggcggaa ccggcggcag cggcagcagc 4200 cccaacgacc ccgtgaccaa catctgccag gccgccgaca agcagctgtt caccctggtg 4260 gagtgggcca agaggattcc ccacttcagc agcctgcccc tggacgacca ggtgatcctg 4320 ctgagggccg gatggaacga gctgctgatc gccagcttca gccacaggag catcgacgtg 4380 agggacggca tcctgctggc caccggcctg cacgtccata ggaacagcgc ccacagcgcc 4440 ggagtgggcg ccatcttcga cag ggtgctg accgagctgg tgagcaagat gagggacatg 4500 aggatggaca agaccgagct gggctgcctg agggccatca tcctgttcaa ccccgaggtg 4560 aggggcctga aaagcgccca ggaggtggag ctgctgaggg agaaggtgta cgccgccctg 4620 gaggagtaca ccaggaccac ccaccccgac gagcccggca gattcgccaa gctgctgctg 4680 aggctgccca gcctgaggag catcggcctg aagtgcctgg agcacctgtt cttcttcagg 4740 ctgatcggcg acgtgcccat cgacaccttc ctgatggaga tgctggagag ccccagcgac 4800 agctgagccg gcaactcgct gtagtaattc cagcgagagg cagagggagc gagcgggcgg 4860 cgggctaggg tggaggagcc cggcgagcag agctgcgctg cgggcgtcct gggaagggag 4920 atccggagcg aatagggggc ttcgcctctg gcccagccct cccgctgatc ccccagccag 4980 cggtgcgcaa ccctagccgc atccacgaaa ctttgcccat agcagcgggc gggcactttg 5040 cactggaact tacaacaccc gagcaaggac gcgactctcc cgacgcgggg aggctattct 5100 gcccatttgg ggacacttcc ccgccgctgc caggacccgc ttctctgaaa 5160 ggctctcctt cttgcgacat ctgcaggctg 5280 gcagctgctt agacgctgga tttttttcgg gtagtggaaa accagcagcc tcccgcgacc 5220 agatctgcca ccatgaagct gctgagcagc atcgagcagg aagaagctga agtgca gcaa ggagaagccc aagtgcgcca agtgcctgaa gaacaactgg 5340 gagtgcagat acagccccaa gaccaagagg agccccctga ccagggccca cctgaccgag 5400 gtggagagca ggctggagag gctggagcag ctgttcctgc tgatcttccc cagggaggac 5460 ctggacatga tcctgaagat ggacagcctg caagacatca aggccctgct gaccggcctg 5520 ttcgtgcagg acaacgtgaa caaggacgcc gtgaccgaca ggctggccag cgtggagacc 5580 gacatgcccc tgaccctgag gcagcacagg atcagcgcca ccagcagcag cgaggagagc 5640 26 - Ο 201207108 agcaacaagg gccagaggca gctgaccgtg agccccgagt ttcccgggat caggcccgag tgcgtggtgc ccgagaccca gtgcgccatg aaaaggaagg agaagaaggc ccagaaggag aaggacaagc tgcccgtgag caccaccacc gtcgatgacc acatgccccc catcatgcag tgcgagcccc ccccccccga ggccgccagg attcacgagg tcgtgcccag gttcctgagc gacaagctgc tggtgaccaa caggcagaag aacatccccc agctgaccgc caaccagcag ttcctgatcg ccaggctgat ctggtatcag gacggctacg agcagcccag cgacgaggac ctgaaaagga tcacccagac ctggcagcag gccgacgacg agaacgagga gagcgacacc cccttcaggc agatcaccga gatgaccatc ctgaccgtgc agctgatcgt ggagttcgcc aagggcctgc ccggattcgc caagatcagc cagcccg acc agatcaccct gctgaaggct tgcagcagcg aggtgatgat gctgagggtg gccaggaggt acgacgccgc cagcgacagc atcctgttcg ccaacaacca ggcttacacc agggacaact acaggaaggc tggcatggcc gaggtgatcg aggacctcct gcacttctgc agatgtatgt acagcatggc cctggacaac atccactacg ccctgctgac cgccgtggtg atcttcagcg acaggcccgg cctggagcag ccccagctgg tggaggagat ccagaggtac tacctgaaca ccctgaggat ctacatcctg aaccagctga gcggcagcgc caggagcagc gtgatctacg gcaagatcct gagcatcctg agcgagctga ggaccctggg aatgcagaac agcaatatgt gtatcagcct gaagctgaag aacaggaagc tgcccccctt cctggaggag atttgggacg tggccgacat gagccacacc cagccccccc ccatcctgga gagccccacc aacctgtgaa tcgattagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc ttaatttgtg aaatttgtga tgctattgct taatttgtaa ccattataag ctgcaataaa caagttaata aaacatttgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatcta gagctcttcc aaatagatct ggaaggtgct gaggtacgat gagacccgca ccaggtgcag accctgcgag tgtggcggta aacatattag gaaccagcct gtgatgctgg atgtgaccga ggagctgagg cccgat cact tggtgctggc ctgcacccgc gctgagtttg gctctagcga tgaagataca gattgaggta ctgaaatgtg tgggcgtggc ttaagggtgg gaaagaatat ataaggtggg ggtcttatgt agttttgtat ctgttttgca gcagccgccg ccgccatgag caccaactcg tttgatggaa gcattgtgag ctcatatttg acaacgcgca tgcccccatg ggccggggtg cgtcagaatg tgatgggctc cagcattgat ggtcgccccg tcctgcccgc aaactctact accttgacct acgagaccgt gtctggaacg ccgttggaga ctgcagcctc cgccgccgct tcagccgctg cagccaccgc ccgcgggatt gtgactgact ttgctttcct gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc cgcgatgaca agttgacggc tcttttggca caattggatt ctttgacccg ggaacttaat gtcgtttctc agcagctgtt ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc cctcccaatg cggtttaaaa cataaataaa aaaccagact ctgtttggat ttggatcaag caagtgtctt gctgtcttta tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg tctcggtcgt tgagggtcct gtgtattttt tccaggacgt ggtaaaggtg actctggatg ttcagataca tgggcataag cccgtctctg gggtggaggt agcaccactg cagagcttca tgctgcgggg tggtgttgta gatgatccag tcgtagcagg agcgctgggc -27 - 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 201207108 10140

gtggtgccta aaaatgtctt tcagtagcaa gctgattgcc aggggcaggc ccttggtgta 7980 agtgtttaca aagcggttaa gctgggatgg gtgcatacgt ggggatatga gatgcatctt 8040 ggactgtatt tttaggttgg ctatgttccc agccatatcc ctccggggat tcatgttgtg 8100 cagaaccacc agcacagtgt atccggtgca cttgggaaat ttgtcatgta gcttagaagg 8160 aaatgcgtgg aagaacttgg agacgccctt gtgacctcca agattttcca tgcattcgtc 8220 cataatgatg gcaatgggcc cacgggcggc ggcctgggcg aagatatttc tgggatcact 8280 aacgtcatag ttgtgttcca ggatgagatc gtcataggcc atttttacaa agcgcgggcg 8340 gagggtgcca gactgcggta taatggttcc atccggccca ggggcgtagt taccctcaca 8400 gatttgcatt tcccacgctt tgagttcaga tggggggatc atgtctacct gcggggcgat 8460 gaagaaaacg gtttccgggg taggggagat cagctgggaa gaaagcaggt tcctgagcag 8520 ctgcgactta ccgcagccgg tgggcccgta aatcacacct attaccgggt gcaactggta 8580 gttaagagag ctgcagctgc cgtcatccct gagcaggggg gccacttcgt taagcatgtc 8640 cctgactcgc atgttttccc tgaccaaatc cgccagaagg cgctcgccgc ccagcgatag 8700 cagttcttgc aaggaagcaa agtttttcaa cggtttgaga ccgtccgccg taggcatgct 8760 tttgag cgtt tgaccaagca gttccaggcg gtcccacagc tcggtcacct gctctacggc 8820 atctcgatcc agcatatctc ctcgtttcgc gggttggggc ggctttcgct gtacggcagt 8880 agtcggtgct cgtccagacg ggccagggtc aigtctttcc acgggcgcag ggtcctcgtc 8940 agcgtagtct gggtcacggt gaaggggtgc gctccgggct gcgcgctggc cagggtgcgc 9000 ttgaggctgg tcctgctggt gctgaagcgc tgccggtctt cgccctgcgc gtcggccagg 9060 tagcatttga ccatggtgtc atagtccagc ccctccgcgg cgtggccctt ggcgcgcagc 9120 ttgcccttgg aggaggcgcc gcacgagggg cagtgcagac ttttgagggc gtagagcttg 9180 ggcgcgagaa ataccgattc cggggagtag gcatccgcgc cgcaggcccc gcagacggtc 9240 tcgcattcca cgagccaggt gagctctggc cgttcggggt caaaaaccag gtttccccca 9300 tgctttttga tgcgtttctt acctctggtt tccatgagcc ggtgtccacg ctcggtgacg 9360 aaaaggctgt ccgtgtcccc gtatacagac ttgagaggcc tgtcctcgac cgatgccctt 9420 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc 9480 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt 9540 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt 9600 attcggaatc t tgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt 9660 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct 9720 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg 9780 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg 9840 acagcttcaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 9900 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 9960 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 10020 tcctgttccg accctgccgc Ttccggata cctgtccgcc tttctccctt cgggaagcgt 10080 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 28 -

201207108 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgittgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt Gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa a &lt;210&gt; 6 &lt;211&gt; 12270 &lt;2Π&gt; DNA &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; Synthetic hIL-12 &lt;400&gt; 6 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg Ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc atcatcaata atatacctta ttttggattg aagccaatat gataatgagg ggg tggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa -29 - 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 11160 11220 11280 11340 11400 11460 11520 11580 11601 60 120 180 240 300 360 420 480 201207108 gtgaaatctg aataattttg tgttactcat agegegtaat atttgtctag ggagatccgg 540 taccgatatc ctagacaacg atgetgaget aactataacg gtcctaaggt agcgaccgcg 600 gagactaggt gtatttatct aagegatege ttaattaagg ccggccgccg caataaaata 660 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac 720 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca 780 agtgcaggtg ccagaacatt tetetatega Taatgcaggt cggagtactg tcctccgagc 840 ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc teegagegga 900 gtactgtcct ccgagcggag tactgtcctc e gageggaga ctcttcgaag gaagaggggc 960 ggggtcgatc gaccccgccc ctcttccttc gaaggaagag gggcggggtc gaagacctag 1020 agggtatata atgggtgcct tagctggtgt gtgagctcat cttcctgtag atcacgcgtc 1080 gaagaaggtg agtaatetta acatgctctt tttttttttt tttgetaate ccttttgtgt 1140 gctgatgtta ggatgacatt tacaacaaat gtttgttcct gacaggaaaa accttgctgg 1200 gtaccttcgt tgccggacac ttcttgtcct ctactttgga aaaaaggaat tgagageege 1260 tagcgccacc atgggtcacc agcagttggt catctcttgg ttttccctgg tttttctggc 1320 atctcccctc gtggccatat gggaactgaa gaaagatgtt tatgtegtag aattggattg 1380 gtatccggat gcccctggag aaatggtggt cctcacctgt gacacccctg aagaagatgg 1440 tatcacctgg accttggacc agagcagtga ggtettagge tctggcaaaa ccctgaccat 1500 ccaagtcaaa gagtttggag atgctggcca gtacacctgt cacaaaggag gcgaggttct 1560 aagccat teg ctcctgctgc ttcacaaaaa ggaagatgga atttggtcca ctgatatttt 1620 aaaggaccag aaagaaccca aaaataagac ett tetaaga tgcgaggcca agaattattc 1680 tggacgtttc acctgctggt ggctgacgac aatcagtact gatttgacat tcagtgtcaa 1740 aageageaga ggctcttctg acc cccaagg ggtgacgtgc ggagctgcta cactctctgc 1800 agagagagtc agaggggaca acaaggagta tgagtactca gtggagtgcc aggaggacag 1860 tgcctgccca gctgctgagg agagtetgee cattgaggtc atggtggatg ccgttcacaa 1920 gctcaagtat gaaaactaca ccagcagctt cttcatcagg gacatcatca aacctgaccc 1980 acccaagaac ttgcagctga agccattaaa gaattetegg caggtggagg tcagctggga 2040 gtaccctgac acctggagta ctccacattc ctacttctcc ctgacattct gcgttcaggt 2100 ccagggcaag agcaagagag aaaagaaaga tagagtette acggacaaga cctcagccac 2160 ggtcatctgc cgcaaaaatg ccagcattag cgtgcgggcc caggaccgct actatagctc 2220 atettggage gaatgggcat ctgtgccctg cagttaggtt gggcgagctc gaattcattg 2280 atcccccggg ctgcaggaat tcgatatcaa getegggate cgaattccgc cccccccccc 2340 cccccccccc taacgt tact ggccgaagcc gettggaata aggccggtgt gcgtttgtct 2400 atatgt tat tt tccaccata t tgeegtet i ttggcaatgt gagggcccgg aaacctggcc 2460 ctgtcttctt gaegageatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc 2520 tgttgaatgt cgtgaaggaa gcagt tcctc tggaagette ttgaagacaa Acaacgtctg 2580 tagcgaccct Ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa 2640 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt 2700 -30 -

201207108 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg gtccagcgcg cagcctcctc cttgtggcta ccctggtcct cctggaccac ctcagtttgg ccagaaacct ccccgtggcc actccagacc caggaatgtt cccatgcctt caccactccc aaaacctgct gagggccgtc agcaacatgc tccagaaggc cagacaaact ctagaatttt acccttgcac ttctgaagag attgatcatg aagatatcac aaaagataaa accagcacag tggaggcctg tttaccattg gaattaacca agaatgagag ttgcctaaat tccagagaga cctctttcat aactaatggg agttgcctgg cctccagaaa gacctctttt atgatggccc tgtgccttag tagtatttat gaagacttga agatgtacca ggtggagttc aagaccatga atgcaaagct tctgatggat cctaagaggc agatctttct agatcaaaac atgctggcag ttattgatga gctgatgcag gccctgaatt tcaacagtga gactgtgcca caaaaatcct cccttgaaga accggatttt tataaaacta aaatcaagct ctgcatactt cttcatgctt tcagaattcg ggcagtgact attgatagag tgatgagcta tctgaatgct tcctaaatcg attgcgcaaa gctttcgcga taggcgagac caatg £ gtgt gtacgtagcg gccgctcgag aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct cgtacggcgt ggtaggtccg aacgaatcca tggattaccc tgttatccct atccggagtt aacctcgagg acttcggaac ttctagaacc agaccgttca gtttaaacgc tcttctcccc ctcgagggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctgggt acgtgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacggca tgcggggggg ggggggggca attggccacc atgggcccc a agaagaaaag gaaggtggcc ccccccaccg acgtgagcct gggcgacgag ctgcacctgg acggcgagga cgtggccatg gcccacgccg acgccctgga cgacttcgac ctggacatgc tgggcgacgg cgacagcccc ggccccggct tcacccccca cgacagcgcc ccctacggcg ccctggacat ggccgacttc gagttcgagc agatgttcac cgacgccctg ggcatcgacg agtacggcgg ccatatggag atgcccgtgg acaggattct ggaggccgaa ctcgccgtgg agcagaaaag cgaccagggc gtggagggcc ccggcggaac cggcggcagc ggcagcagcc ccaacgaccc cgtgaccaac atctgccagg ccgccgacaa gcagctgttc accctggtgg agtgggccaa gaggattccc cacttcagca gcctgcccct ggacgaccag -31 - 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 201207108 gtgatcctgc tgagggccgg atggaacgag ctgctgatcg ccagcttcag ccacaggagc 5040 atcgacgtga gggacggcat cctgctggcc Accggcctgc acgtccatag gaacagcgcc 5100 cacagcgccg gagtgggcgc catcttcgac agggtgctga ccgagctggt gagcaagatg 5160 agggacatga ggatggacaa gaccgagctg ggctgcctga gggccatcat cctgttc aac 5220 cccgaggtga ggggcctgaa aagcgcccag gaggtggagc tgctgaggga gaaggtgtac 5280 gccgccctgg aggagtacac caggaccacc caccccgacg agcccggcag attcgccaag 5340 ctgctgctga ggctgcccag cctgaggagc atcggcctga agtgcctgga gcacctgttc 5400 ttcttcaggc tgatcggcga cgtgcccatc gacaccttcc tgatggagat gctggagagc 5460 cccagcgaca gctgagccgg caactcgctg tagtaattcc agcgagaggc agagggagcg 5520 agcgggcggc gggctagggt ggaggagccc ggcgagcaga gctgcgctgc gggcgtcctg 5580 ggaagggaga tccggagcga atagggggct tcgcctctgg cccagccctc ccgctgatcc 5640 cccagccagc ggtgcgcaac cctagccgca tccacgaaac tttgcccata gcagcgggcg 5700 ggcactttgc actggaactt acaacacccg agcaaggacg cgactctccc gacgcgggga 5760 ggctattctg cccatttggg gacacttccc cgccgctgcc aggacccgct tctctgaaag 5820 gctctccttg cagctgctta gacgctggat ttttttcggg tagtggaaaa ccagcagcct 5880 cccgcgacca gatctgccac catgaagctg ctgagcagca tcgagcaggc ttgcgacatc 5940 tgcaggctga agaagctgaa gtgcagcaag gagaagccca agtgcgccaa gtgcctgaag 6000 aacaactggg agtgcagata cagccccaag accaagagga gccccctgac cagggcccac 60 60 ctgaccgagg tggagagcag gctggagagg ctggagcagc tgttcctgct 6120 gatcttcccc gcccgtgagc accaccaccg tcgatgacca agggaggacc tggacatgat cctgaagatg gacagcctgc aagacatcaa ggccctgctg 6180 accggcctgt tcgtgcagga caacgtgaac aaggacgccg tgaccgacag gctggccagc 6240 gtggagaccg acatgcccct gaccctgagg cagcacagga tcagcgccac cagcagcagc 6300 gaggagagca gcaacaaggg ccagaggcag ctgaccgtga gccccgagtt tcccgggatc 6360 aggcccgagt gcgtggtgcc cgagacccag tgcgccatga aaaggaagga gaagaaggcc 6420 cagaaggaga aggacaagct catgcccccc 6480 atcatgcagt gcgagccccc cccccccgag gccgccagga ttcacgaggt cgtgcccagg 6540 ttcctgagcg acaagctgct ggtgaccaac aggcagaaga acatccccca gctgaccgcc 6600 aaccagcagt tcctgatcgc caggctgatc tggtatcagg acggctacga gcagcccagc 6660 gacgaggacc tgaaaaggat cacccagacc tggcagcagg ccgacgacga gaacgaggag 6720 agcgacaccc ccttcaggca gatcaccgag atgaccatcc tgaccgtgca gctgatcgtg 6780 gagttcgcca agggcctgcc cggattcgcc aagatcagcc agcccgacca gatcaccctg 6840 ctgaaggctt gcagcagcga ggtgatgatg cig ^ gggigg ccaggaggta cgacgccgcc 6900 agc gacagca tcctgttcgc caacaaccag gcttacacca gggacaacta caggaaggct 6960 ggcatggccg aggtgatcga ggacctcctg cacttctgca gatgtatgta cagcatggcc 7020 ctggacaaca tccactacgc cctgctgacc gccgtggtga tcttcagcga caggcccggc 7080 ctggagcagc cccagctggt ggaggagatc cagaggtact acctgaacac cctgaggatc 7140 tacatcctga accagctgag cggcagcgcc aggagcagcg tgatctacgg caagatcctg 7200 -32 -

201207108 agcatcctga gcgagctgag gaccctggga atgcagaaca gcaatatgtg tatcagcctg aagctgaaga acaggaagct gccccccttc ctggaggaga tttgggacgt ggccgacatg agccacaccc agcccccccc catcctggag agccccacca acctgtgaat cgattagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct taatttgtga aatttgtgat gctattgctt aatttgtaac cattataagc tgcaataaac aagttaataa aacatttgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatctag agctcttcca aatagatctg gaaggtgctg aggtacgatg agacccgcac caggtgcaga ccctgcgagt gtggcggtaa acatattagg aaccagcctg tgatgctgga tgtgaccgag gagctgaggc ccgatcactt ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat gaagatacag attgaggtac tgaaatgtgt gggcgtggct taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc cgcgggattg tgactgactt tgctttcctg agcccgcttg caagcagtgc agcttcccgt tcatccgccc gcgatgacaa gttgacggct cttttggcac aattggattc tttgacccgg gaacttaatg tcgtttctca gcagctgttg gatctgcgcc agcaggtttc tgccctgaag gcttcctccc ctcccaatgc ggtttaaaac ataaataaaa aaccagactc tgtttggatt tggatcaagc aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg gtaggcccgg gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg gtaaaggtga ctctggatgt tcagatacat gggcataagc ccgtctctgg ggtggaggta gcaccactgc agagcttcat gctgcggggt ggtgttgtag atgatccagt cgtagcagga gcgctgggcg tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc cttggtgtaa gtgtttacaa agcggttaag ctgggatggg tgcatacgtg gggatatgag atgcatcttg gactgtattt ttaggttggc tatgttccca gccatatccc tccggggatt catgttgtgc agaaccacca gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag cttagaagga aatgcgtgga agaacttgga gacgcccttg tgacctccaa gattttccat gcattcgtcc ataatgatgg caatgggccc acgggcggcg gcctgggcga agatatttct gggatcacta acgtcatagt tgtgttccag gatgagatcg tcataggcca tttttacaaa gcgcgggcgg agggtgccag actgcggtat aatggttcca tccggcccag gggcgtagtt accctcacag atttgcattt cccacgcttt gagttcagat ggggggatca tgtctacctg cggggcgatg aagaaaacgg tttccggggt aggggagatc agctgggaag aaagcaggtt cctgagcagc tgcgacttac cgcagccggt gggcccgtaa atcacaccta ttaccgggtg caactggtag ttaagagagc tgcagctgcc gtcatccctg agcagggggg ccacttcgtt aagcatgtcc ctgactcgca tgttttccct gaccaaatcc gccagaaggc gctcgccgcc cagcgatagc agttcttgca aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt aggcatgctt ttgagcgttt gaccaagcag ttccaggcgg tcccacagct cggtcacctg -33 - 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 201207108 ctctacggca tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg 9540 tacggcagta gtcggtgctc gtccagacgg gccagggtca Tgtctttcca cgggcgcagg 9600 gtcctcgtca gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc 9660 agggtgcgct tgaggctggt cctgctggtg ctgaagcgct gccggtcttc gccct gcgcg 9720 tcggccaggt agcatttgac catggtgtca tagtccagcc cctccgcggc gtggcccttg 9780 gcgcgcagct tgcccttgga ggaggcgccg cacgaggggc agtgcagact tttgagggcg 9840 tagagcttgg gcgcgagaaa taccgattcc ggggagtagg catccgcgcc gcaggccccg 9900 cagacggtct cgcattccac gagccaggtg agctctggcc gttcggggtc aaaaaccagg 9960 tttcccccat gctttttgat gcgtttctta cctctggttt ccatgagccg gtgtccacgc 10020 tcggtgacga aaaggctgtc cgtgtccccg tatacagact tgagaggcct gtcctcgacc 10080 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat 10140 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc 10200 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc 10260 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac 10320 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta 10380 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc 10440 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga 10500 ccatcaggga cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 10560 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 10620 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 10680 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 10740 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 10800 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 10860 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 10920 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 10980 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 11040 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 11100 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 11160 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 11220 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 11280 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 11340 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 11400 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 11460 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 11520 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 11580 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 13640 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 11700 -34 -

201207108 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aaiaccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc Ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa &lt;210> 7 &lt;211&gt; 10404 &lt;212&gt; DNA &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; Synthetic mIL-21 &lt;400>7 taaacaaata ggggttccgc gcacatttcc Ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc atcatcaata atatacctta ttttggattg aagccaat at gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag ggagatccgg taccgatatc ctagacaacg atgctgagct aactataacg gtcctaaggt agcgaccgcg gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taatgcaggt cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggaga ctcttcgaag gaagaggggc ggggtcgatc gaccccgccc ctcttccttc gaaggaagag gggcggggtc gaagacctag agggtatata atgggtgcct tagctggtgt gtgagctcat cttcctgtag atcacgcgic gaagaaggtg agtaatctta acatgctctt tttttttttt tttgcta atc ccttttgtgt gctgatgtta ggatgacatt tacaacaaat gtttgttcct gacaggaaaa accttgctgg gtaccttcgt tgccggacac ttcttgtcct ctactttgga aaaaaggaat tgagagccgc tagcccacca tggagaggac cctggtgtgc ctggtggtga tcttcctggg caccgtggcc cacaagagca gcccccaggg acccgacagg ctgctgatcc ggctgagaca cctgatcgac -35 - 11760 11820 11880 11940 12000 12060 12120 12180 12240 12270 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 201207108 atcgtggagc agctgaagat ttacgagaac gacctggacc ccgagctgct gtccgccccc 1440 caggacgtga agggccactg cgagcacgcc gccttcgcct gcttccagaa ggccaagctg 1500 aagcccagca accccggcaa caacaagacc ttcatcatcg acctggtggc ccagctgaga 1560 aggaggctgc ccgccaggag gggcggcaag aagcagaagc acatcgccaa gtgccccagc 1620 tgcgacagct acgagaagcg gacccccaag gagttcctgg agaggctgaa gtggctgctg 1680 caaaagatga tccaccagca Cctgagctga atcgattgcg caaagct t tc gcgataggcg 1740 agaccaatgg gtgtgtacgt agcggccgct cgagaacttg tttattgcag cttataatgg 1800 ttacaaataa agcaatagca tcacaaattt cacaaata aa gcattttttt cactgcattc 1860 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctcgtacg gcgtggtagg 1920 tccgaacgaa tccatggatt accctgttat ccctatccgg agttaacctc gaggacttcg 1980 gaacttctag aaccagaccg ttcagtttaa acgctcttct ccccctcgag ggcctccgcg 2040 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga 2100 cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg 2160 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt 2220 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc 2280 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa 2340 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt 2400 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct gggtacgtgc 2460 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat 2520 ttgggtcaat atgtaatt tt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt 2580 ttttggcttt tttgttagac ggcatgcggg gggggggggg ggcaattggc caccatgggc 2640 cccaagaaga aaaggaaggt ggcccccccc accgacgtga gcctgggcga cgagctgcac 2700 ctggacggcg aggacgtggc catggcccac gccgacgccc tggacgactt cgacctggac 2760 atgctgggcg acggcgacag ccccggcccc ggcttcaccc cccacgacag cgccccctac 2820 ggcgccctgg acatggccga cttcgagttc gagcagatgt tcaccgacgc cctgggcatc 2880 gacgagtacg gcggccatat ggagatgccc gtggacagga ttctggaggc cgaactcgcc 2940 gtggagcaga aaagcgacca gggcgtggag ggccccggcg gaaccggcgg cagcggcagc 3000 agccccaacg accccgtgac caacatctgc caggccgccg acaagcagct gttcaccctg 3060 gtggagtggg ccaagaggat tccccacttc agcagcctgc ccctggacga ccaggtgatc 3120 ctgctgaggg ccggatggaa cgagctgctg atcgccagct tcagccacag gagcatcgac 3180 gtgagggacg gcatcctgct ggccaccggc ctgcacgtcc ataggaacag cgcccacagc 3240 gccggagtgg gcgccatctt cgacagggtg ctgaccgagc tggtgagcaa gatgagggac 3300 atgaggatgg acaagaccga gctgggctgc ctgagggcca tcatcctgtt caaccccgag 3360 gtgaggggcc tgaaaagcgc ccaggaggtg gagctgctga gggagaaggt gtacgccgcc 3420 ctggaggagt acaccaggac cacccacccc gacgagcccg gcagattcgc caagctgctg 3480 ctgaggctgc ccagcctgag ga Gcatcggc ctgaagtgcc tggagcacct gttcttcttc 3540 aggctgatcg gcgacgtgcc catcgacacc ttcctgatgg agatgctgga gagccccagc 3600 -36 - 201207108

gacagctgag cggcgggcta gagatccgga cagcggtgcg ttgcactgga tctgcccatt cttgcagctg accagatctg ctgaagaagc tgggagtgca gaggtggaga gacctggaca ctgttcgtgc accgacatgc agcagcaaca gagtgcgtgg gagaaggaca cagtgcgagc agcgacaagc cagttcctga gacctgaaaa acccccttca gccaagggcc gcttgcagca agcatcctgt gccgaggtga aacatccact cagccccagc ctgaaccagc ctgagcgagc aagaacagga acccagcccc gatacattga gtgaaatttg ataaaacatt aagcaagtaa gctgaggtac taggaaccag ccggcaactc gggtggagga gcgaataggg caaccctagc acttacaaca tggggacact cttagacgct ccaccatgaa tgaagtgcag gatacagccc gcaggctgga tgatcctgaa aggacaacgt ccctgaccct agggccagag tgcccgagac agctgcccgt cccccccccc tgctggtgac tcgccaggct ggatcaccca ggcagatcac tgcccggatt gcgaggtgat tcgccaacaa tcgaggacct acgccctgct tggtggagga tgagcggcag tgaggaccct agctgccccc cccccatcct tgagtttgga tgatgctatt tgcattcatt aacctctaca gatgagaccc cctgtgatgc gctgtagtaa gcccggcgag ggcttcgcct cgcatccacg cccgagcaag tccccgccgc ggattttttt gctgctgagc caaggagaag caagaccaag gaggctggag gatggacagc gaacaaggac gaggcagcac gcagctgacc ccagtgcgcc gagcaccacc cgaggccgcc caacaggcag gatctggtat gacctggcag cgagatgacc cgccaagatc gatgctgagg ccaggcttac cctgcacttc gaccgccgtg gatccagagg cgccaggagc gggaatgcag cttcctggag ggagagcccc caaaccacaa gcttaatttg ttatgtttca aatgtggtat gcaccaggtg tggatgtgac ttccagcgag cagagctgcg ctggcccagc aaactttgcc gacgcgactc tgccaggacc cgggtagtgg agcatcgagc cccaagtgcg aggagccccc cagctgttcc ctgcaagaca gccgtgaccg aggatcagcg gtgagccccg atgaaaagga accgtcgatg aggattcacg aagaacatcc caggacggct caggccgacg atcctgaccg agccagcccg gtggccagga accagggaca tgcagatgta gtgatcttca tactacctga agcgtgatct aacagcaata gagatttggg accaacctgt ctagaatgca taaccattat ggttcagggg ctagagctct cagaccctgc cgaggagctg aggcagaggg ctgcgggcgt cctcccgctg catagcagcg tcccgacgcg cgcttctctg aaaaccagca aggcttgcga ccaagtgcct tgaccagggc tgctgatctt tcaaggccct acaggctggc ccaccagcag agtttcccgg aggagaagaa accacatgcc aggtcgtgcc cccagctgac acgagcagcc acgagaacga tgcagctgat accagatcac ggtacgacgc actacaggaa tgtacagcat gcgacaggcc acaccctgag acggcaagat tgtgtatca g acgtggccga gaatcgatta gtgaaaaaaa aagctgcaat gagatgtggg tccaaataga gagtgtggcg aggcccgatc -37 - agcgagcggg cctgggaagg atcccccagc ggcgggcact gggaggctat aaaggctctc gcctcccgcg catctgcagg gaagaacaac ccacctgacc ccccagggag gctgaccggc cagcgtggag cagcgaggag gatcaggccc ggcccagaag ccccatcatg caggttcctg cgccaaccag cagcgacgag ggagagcgac cgtggagttc cctgctgaag cgccagcgac ggctggcatg ggccctggac cggcctggag gatctacatc cctgagcatc cctgaagctg catgagccac gacatgataa tgcttaattt aaacaagtta aggtttttta tctggaaggt gtaaacatat acttggtgct 3660 3720 3780 3840 3900 3960 4020 4080 4240 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 201207108 ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagattgag gtactgaaat 5940 gtgtgggcgt ggcttaaggg tgggaaagaa tatataaggt Gggggtctta tgtagttttg 6000 tatctgtttt gcagcagccg ccgccgccat gagcaccaac tcgtttgatg gaagcattgt 6060 gagctcatat ttgacaacgc gcatgccccc atgggccggg gtgcgtcaga atgtgatggg 6120 ct ccagcatt gatggtcgcc ccgtcctgcc cgcaaactct actaccttga cctacgagac 6180 cgtgtctgga acgccgttgg agactgcagc ctccgccgcc gcttcagccg ctgcagccac 6240 cgcccgcggg attgtgactg actttgcttt cctgagcccg cttgcaagca gtgcagcttc 6300 ccgttcatcc gcccgcgatg acaagttgac ggctcttttg gcacaattgg attctttgac 6360 ccgggaactt aatgtcgttt ctcagcagct gttggatctg cgccagcagg tttctgccct 6420 gaaggcttcc tcccctccca atgcggttta aaacataaat aaaaaaccag actctgtttg 6480 gatttggatc aagcaagtgt cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc 6540 ccgggaccag cggtctcggt cgttgagggt cctgtgtatt ttttccagga cgtggtaaag 6600 gtgactctgg atgttcagat acatgggcat aagcccgtct ctggggtgga ggtagcacca 6660 ctgcagagct tcatgctgcg gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg 6720 ggcgtggtgc ctaaaaatgt ctttcagtag caagctgatt gccaggggca ggcccttggt 6780 gtaagtgttt acaaagcggt taagctggga tgggtgcata cgtggggata tgagatgcat 6840 cttggactgt atttttaggt tggctatgtt cccagccata tccctccggg gattcatgtt 6900 gtgcagaacc accagcacag tgtatccggt gcacttggga aatttgtcat gtagcttaga 6960 aggaaatg cg tggaagaact tggagacgcc cttgtgacct ccaagatttt ccatgcattc 7020 gtccataatg atggcaatgg gcccacgggc ggcggcctgg gcgaagatat ttctgggatc 7080 actaacgtca tagttgtgtt ccaggatgag atcgtcatag gccattttta caaagcgcgg 7140 gcggagggtg ccagactgcg gtataatggt tccatccggc ccaggggcgt agttaccctc 7200 acagatttgc atttcccacg ctttgagttc agatgggggg atcatgtcta cctgcggggc 7260 gatgaagaaa acggtttccg gggtagggga gatcagctgg gaagaaagca ggttcctgag 7320 cagctgcgac ttaccgcagc cggtgggccc gtaaatcaca cctattaccg ggtgcaactg 7380 gtagttaaga gagctgcagc tgccgtcatc cctgagcagg ggggccactt cgttaagcat 7440 gtccctgact cgcatgtttt ccctgaccaa atccgccaga aggcgctcgc cgcccagcga 7500 tagcagttct tgcaaggaag caaagttttt caacggtttg agaccgtccg ccgtaggcat 7560 gcttttgagc gtttgaccaa gcagttccag gcggtcccac agctcggtca cctgctctac 7620 ggcatctcga tccagcatat ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc 7680 agtagtcggt gctcgtccag acgggccagg gtcatgtctt tccacgggcg cagggtcctc 7740 gtcagcgtag tctgggtcac ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg 7800 cgcttgaggc tgg tcctgct ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc 7860 aggtagcatt tgaccatggt gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc 7920 agcttgccct tggaggaggc gccgcacgag gggcagtgca gacttttgag ggcgtagagc 7980 ttgggcgcga gaaataccga ttccggggag taggcatccg cgccgcaggc cccgcagacg 8040 gtctcgcatt ccacgagcca ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc 8100 -38 - 201207108

ccatgctttt tgatgcgttt cttacctctg gtttccatga gccggtgtcc acgctcggtg 8160 acgaaaaggc tgtccgtgtc cccgtataca gacttgagag gcctgtcctc gaccgatgcc 8220 cttgagagcc ttcaacccag tcagctccit ccggtgggcg cggggcatga ctatcgtcgc 8280 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg 8340 ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc 8400 ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg 8460 tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt 8520 gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg 8580 cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca 8640 gggacagctt caaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 8700 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 8760 gcgaaacccg acaggactat aaagaiacca ggcgtttccc cctggaagct ccctcgtgcg 8820 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 8880 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgct c 8940 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 9000 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 9060 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 9120 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 9180 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 9240 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 9300 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 9360 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 9420 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 9480 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 9540 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 9600 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 9660 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 9720 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca t tgctgcagg 9780 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 9840 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 9900 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 9960 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 10020 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg 10080 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 10140 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tcggtcctcc 10200 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 10260 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 10320 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca -39 - tgagcggata 10380 201207108 catatttgaa tgtatttaga aaaa &lt; 210> 8 &lt; 211 &gt; 10452 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt;220&gt;&lt;223&gt; Synthetic hIL-21 &lt;400> 8 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac catiattatc atgacattaa cc tataaaaa taggcgtatc acgaggccct ttcgtcttca atcatcaata attaatcgca ccggtatcta tgtcgggtgc ggagaaagag gtaatgaaat ggcagctagc ggagatccgg taccgatatc ctagacaacg atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat atttgtctag atgctgagct aactataacg gtcctaaggt agcgaccgcg gagactaggt gtatttatct aagcgatcgc ttaattaagg ccggccgccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taatgcaggt cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggaga ctcttcgaag gaagaggggc ggggtcgatc gaccccgccc ctcttccttc g aaggaagag gggcggggtc gaagacctag agggtatata atgggtgcct tagctggtgt gtgagctcat cttcctgtag atcacgcgtc gaagaaggtg agtaatctta acatgctctt tttttttttt tttgctaatc ccttttgtgt gctgatgtta ggatgacatt tacaacaaat gtttgttcct gacaggaaaa accttgctgg gtaccttcgt tgccggacac ttcttgtcct ctactttgga aaaaaggaat tgagagccgc tagcccacca tgagaagcag ccccggcaac atggagagaa tcgtgatctg cctgatggtg atcttcctgg gcaccctggt gcataagagc agcagccagg gccaggacag acacatgatc cgcatgagac agctgatcga catcgtggac cagctgaaga actacgtgaa cgacctggtg cccgagttcc tgcccgcccc cgaggacgtg gagaccaact gcgagtggag cgccttcagc tgcttccaga aggcccagct gaagtccgcc aacaccggca acaacgagag aatcatcaac gtgagcatca agaagctgaa gcggaagccc cccagcacca acgccggaag aagacagaag cacagactga cctgtcccag ctgcgacagc tacgagaaga agccccccaa ggagttcctg gagagattca agagcctgct gcaaaagatg atccaccagc acctgagcag cagaacccac ggcagcgagg acagctgaat cgattgcgca aagctttcgc gataggcgag accaatgggt gtgtacgtag cggccgctcg agaacttgtt tattgcagct tataatggtt acaaataaag 10404 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 -40 - 201207108

caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 1920 gtccaaactc atcaatgtat cttatcatgt ctcgtacggc gtggtaggtc cgaacgaatc 1980 catggattac cctgttatcc ctatccggag ttaacctcga ggacttcgga acttctagaa 2040 ccagaccgtt cagtttaaac gctcttctcc ccctcgaggg cctccgcgcc gggttttggc 2100 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag 2160 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc 2220 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag 2280 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga 2340 ttctgcggag ggatctccgt ggsgcggtga acgccgatga ttatataagg acgcgccggg 2400 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg 2460 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg gtacgtgcgc tcggggttgg 2520 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat 2580 gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt 2640 tgttagacgg catgcggggg gggggggggg caattggcca ccatgggccc caagaagaa a 2700 aggaaggtgg ccccccccac cgacgtgagc ctgggcgacg agctgcacct ggacggcgag 2760 gacgtggcca tggcccacgc cgacgccctg gacgacttcg acctggacat gctgggcgac 2820 ggcgacagcc ccggccccgg cttcaccccc cacgacagcg ccccctacgg cgccctggac 2880 atggccgact tcgagttcga gcagatgttc accgacgccc tgggcatcga cgagtacggc 2940 ggccatatgg agatgcccgt ggacaggatt ctggaggccg aactcgccgt ggagcagaaa 3000 agcgaccagg gcgtggaggg ccccggcgga accggcggca gcggcagcag ccccaacgac 3060 cccgtgacca acatctgcca ggccgccgac aagcagctgt tcaccctggt ggagtgggcc 3120 aagaggattc cccacttcag cagcctgccc ctggacgacc aggtgatcct gctgagggcc 3180 ggatggaacg agctgctgat cgccagcttc agccacagga gcatcgacgt gagggacggc 3240 atcctgctgg ccaccggcct gcacgtccat aggaacagcg cccacagcgc cggagtgggc 3300 gccatcttcg acagggtgct gaccgagctg gtgagcaaga tgagggacat gaggatggac 3360 aagaccgagc tgggctgcct gagggccatc atcctgttca accccgaggt gaggggcctg 3420 aaaagcgccc aggaggtgga gctgctgagg gagaaggtgt acgccgccct ggaggagtac 3480 accaggacca cccaccccga cgagcccggc agattcgcca agctgctgct g aggctgccc 3540 agcctgagga gcatcggcct gaagtgcctg gagcacctgt tcttcttcag gctgatcggc 3600 gacgtgccca tcgacacctt cctgatggag atgctggaga gccccagcga cagctgagcc 3660 ggcaactcgc tgtagtaatt ccagcgagag gcagagggag cgagcgggcg gcgggctagg 3720 gtggaggagc ccggcgagca gagctgcgct gcgggcgtcc tgggaaggga gatccggagc 3780 gaataggggg cttcgcctct ggcccagccc tcccgctgat cccccagcca gcggtgcgca 3840 accctagccg catccacgaa actttgccca tagcagcggg cgggcacttt gcactggaac 3900 ttacaacacc cgagcaagga cgcgactctc ccgacgcggg gaggctattc tgcccatttg 3960 gggacacttc cccgccgctg ccaggacccg cttctctgaa aggctctcct tgcagctgct 4020 tagacgctgg atttttttcg ggtagtggaa aaccagcagc ctcccgcgac cagatctgcc 4080 accatgaagc tgctgagcag catcgagcag gcttgcgaca tctgcaggct -41 - gaagaagctg 4140 201207108 aagtgcagca aggagaagcc caagtgcgcc aagtgcctga agaacaactg ggagtgcaga 4200 tacagcccca agaccaagag gagccccctg accagggccc acctgaccga ggtggagagc 4260 aggctggaga ggctggagca gctgttcctg ctgatcttcc ccagggagga cctggacatg 4320 atcctgaaga tggacagcct gcaagacatcaaggccctgc tgaccggcct gttcgtgcag 4380 gacaacgtga acaaggacgc cgtgaccgac aggctggcca gcgtggagac cgacatgccc 4440 ctgaccctga ggcagcacag gatcagcgcc accagcagca gcgaggagag cagcaacaag 4500 ggccagaggc agctgaccgt gagccccgag tttcccggga tcaggcccga gtgcgtggtg 4560 cccgagaccc agtgcgccat gaaaaggaag gagaagaagg cccagaagga gaaggacaag 4620 ctgcccgtga gcaccaccac cgtcgatgac cacatgcccc ccatcatgca gtgcgagccc 4680 cccccccccg aggccgccag gattcacgag gtcgtgccca ggttcctgag cgacaagctg 4740 ctggtgacca acaggcagaa gaacatcccc cagctgaccg ccaaccagca gttcctgatc 4800 gccaggctga tctggtatca ggacggctac gagcagccca gcgacgagga cctgaaaagg 4860 atcacccaga cctggcagca ggccgacgac gagaacgagg agagcgacac ccccttcagg 4920 cagatcaccg agatgaccat cctgaccgtg cagctgatcg tggagttcgc caagggcctg 4980 cccggattcg ccaagatcag ccagcccgac cagatcaccc tgctgaaggc ttgcagcagc 5040 gaggtgatga tgctgagggt ggccaggagg tacgacgccg ccagcgacag catcctgttc 5100 gccaacaacc aggcttacac cagggacaac tacaggaagg ctggcatggc cgaggtgatc 5160 gaggacctcc tgcacttctg cag atgtatg tacagcatgg ccctggacaa catccactac 5220 gccctgctga ccgccgtggt gatcttcagc gacaggcccg gcctggagca gccccagctg 5280 gtggaggaga tccagaggta ctacctgaac accctgagga tctacatcct gaaccagctg 5340 agcggcagcg ccaggagcag cgtgatctac ggcaagatcc tgagcatcct gagcgagctg 5400 aggaccctgg gaatgcagaa cagcaatatg tgtatcagcc tgaagctgaa gaacaggaag 5460 ctgcccccct tcctggagga gatttgggac gtggccgaca tgagccacac ccagcccccc 5520 cccatcctgg agagccccac caacctgtga atcgattaga catgataaga tacattgatg 5580 agtttggaca aaccacaact agaatgcagt gaaaaaaatg cttaatttgt gaaatttgtg 5640 atgctattgc ttaatttgta accattataa gctgcaataa acaagttaat aaaacatttg 5700 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa 5760 cctctacaaa tgtggtatct agagctcttc caaatagatc tggaaggtgc tgaggtacga 5820 tgagacccgc accaggtgca gaccctgcga gtgtggcggt aaacatatta ggaaccagcc 5880 tgtgatgctg gatgtgaccg aggagctgag gcccgatcac ttggtgctgg cctgcacccg 5940 cgctgagttt ggctctagcg atgaagatac agattgaggt actgaaatgt gtgggcgtgg 6000 cttaagggtg ggaaag aata tataaggtgg gggtcttatg tagttttgta tctgttttgc 6060 agcagccgcc gccgccatga gcaccaactc gtttgatgga agcattgtga gctcatattt 6120 gacaacgcgc atgcccccat gggccggggt gcgtcagaat gtgatgggct ccagcattga 6180 tggtcgcccc gtcctgcccg caaactctac taccttgacc tacgagaccg tgtctggaac 6240 gccgttggag actgcagcct ccgccgccgc ttcagccgct gcagccaccg cccgcgggat 6300 tgtgactgac tttgctttcc tgagcccgct tgcaagcagt gcagct tccc gt tcatccgc 6360 • 42 - 201207108

ccgcgatgac tgtcgtttct ccctcccaat gcaagtgtct gtctcggtcg gttcagatac atgctgcggg aaaaatgtct aaagcggtta ttttaggttg cagcacagtg gaagaacttg ggcaatgggc gttgtgttcc agactgcggt ttcccacgct ggtttccggg accgcagccg gctgcagctg catgttttcc caaggaagca ttgaccaagc cagcatatct tcgtccagac tgggtcacgg gtcctgctgg accatggtgt gaggaggcgc aataccgatt acgagccagg atgcgtttct tccgtgtccc caacccagtc tgtcttcttt cgaggaccgc cttgcacgcc gcaggccatt gacgcgaggc aagttgacgg cagcagctgt gcggtttaaa tgctgtcttt ttgagggtcc atgggcataa gtggtgttgt ttcagtagca agctgggatg gctatgttcc tatccggtgc gagacgccct ccacgggcgg aggatgagat ataatggttc ttgagttcag gtaggggaga gtgggcccgt ccgtcatccc ctgaccaaat aagtttttca agttccaggc cctcgtttcg gggccagggt tgaaggggtg tgctgaagcg catagtccag cgcacgaggg ccggggagta tgagctctgg tacctctggt cgtatacaga agctccttcc atcatgcaac tttcgctgga ctcgctcaag atcgccggca tggatggcct ctcttttggc tggatctgcg acataaataa atttaggggt tgtgtatttt gcccgtctct agatgatcca agctgattgc ggtgcatacg cagccatatc acttgggaaa tgtgacctcc cggcctgggc cgtcataggc catccggccc atggggggat tcagctggga aaatcacacc tgagcagggg ccgccagaag acggtttgag ggtcccacag cgggttgggg catgtctttc cgctccgggc ctgccggtct cccctccgcg gcagtgcaga ggcatccgcg ccgttcgggg ttccatgagc cttgagaggc ggtgggcgcg tcgtaggaca gcgcgacgat ccttcgtcac tggcggccga tccccattat acaattggat ccagcaggtt aaaaccagac tttgcgcgcg ttccaggacg ggggtggagg gtcgtagcag caggggcagg tggggatatg cctccgggga tttgtcatgt aagattttcc gaagatattt catttttaca aggggcgtag catgtctacc agaaagcagg tattaccggg ggccacttcg gcgctcgccg accgtccgcc ctcggtcacc cggctttcgc cacgggcgca tgcgcgctgg tcgccctgcg gcgtggccct cttttgaggg ccgcaggccc tcaaaaacca cggtgtccac ctgtcctcga gggcatgact ggtgccggca gatcggcctg tggtcccgcc cgcgctgggc gattcttctc tctttgaccc tctgccctga tctgtttgga cggtaggccc tggtaaaggt tagcaccact gagcgctggg cccttggtgt agatgcatct ttcatgttgt agcttagaag atgcattcgt ctgggatcac aagcgcgggc ttaccctcac tgcggggcga ttcctgagca tgcaactggt ttaagcatgt cccagcgata gtaggcatgc tgctctacgg tgtacggcag gggtcctcgt ccagggtgcg cgtcggccag tggcgcgcag cgtagagctt cgcagacggt ggtttcccc c gctcggtgac ccgatgccct atcgtcgccg gcgctctggg tcgcttgcgg accaaacgtt tacgtcttgc gcttccggcg -43 - gggaacttaa aggcttcctc tttggatcaa gggaccagcg gactctggat gcagagcttc cgtggtgcct aagtgtttac tggactgtat gcagaaccac gaaatgcgtg ccataatgat taacgtcata ggagggtgcc agat ttgcat tgaagaaaac gctgcgactt agttaagaga ccctgactcg gcagttcttg ttttgagcgt catctcgatc tagtcggtgc cagcgtagtc cttgaggctg gtagcatttg cttgcccttg g £ gcgcgaga ctcgcattcc atgctttttg gaaaaggctg tgagagcctt cacttatgac tcattttcgg tattcggaat Tcggcgagaa tggcgttcgc gcatcgggat 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 201207108 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca 8700 aggccagcaa Aaggccagga accgiaaaaa ggccgcgttg ctggcgttti tccataggct 8760 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 8820 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 8880 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 8940 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 9000 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 9060 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 9120 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 9180 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 9240 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 9300 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 9360 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 9420 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 9480 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 9540 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 9600 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 9660 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 9720 tcctgc aact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 9780 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc 9840 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 9900 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 9960 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 10020 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 10080 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc 10140 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 10200 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 10260 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 10320 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 10380 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 10440 tatttagaaa aa 10452 &lt; 210 &gt; 9 &lt; 211 &gt; 489 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Synthetic mIL-21 &lt;220&gt;

&lt;221&gt; CDS -44 - 48 48

201207108 &lt;222&gt; (1)..(489) &lt;400&gt; 9 atg aag ate ctg aag ccc tac atg agg accac ate age tgt tac

Met Lys lie Leu Lys Pro Tyr Met Arg Asn Thr Ser lie Ser Cys Tyr 15 10 15 ctg tgc ttc ctg ctg aac age cac ttc ctg acc gag gee gga ate cac

Leu Cys Phe Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly lie His 20 25 30 gtc ttc ate ctg ggc tgc gtg age gtg ggc ctg ccc aag acc gag gee

Val Phe lie Leu Gly Cys Val Ser Val Gly Leu Pro Lys Thr Glu Ala 35 40 45 aac tgg ate gac gtg agg tac gac ctg gag aag ate gag age ctg ate

Asn Trp lie Asp Val Arg Tyr Asp Leu Glu Lys lie Glu Ser Leu lie 50 55 60 cag age ate cac ate gac acc acc ctg tac acc gac age gac ttc cac Gin Ser lie His lie Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His 65 70 75 80 ccc age tgc aag gtg acc gee atg aac tgc ttc ctg ctg gag ctg caa

Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gin 85 90 95 gtg ate ctg cac gag tac age aac atg acc ctg aac gag acc gtg agg

Val lie Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg 100 105 110 aac gtg ctg tac ctg get aac age acc ctg age age aac aag aac gtg

Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val 115 120 125 gee gag age ggc tgc aag gag tgt gag gag ctg gag gag aag acc ttc

Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe 130 135 140 acc gag ttc etc cag age ttc ate agg ate gtg cag atg ttc ate aac

Thr Glu Phe Leu Gin Ser Phe lie Arg lie Val Gin Met Phe lie Asn 145 150 155 160 acc age tga Thr Ser &lt;210&gt; 10 &lt;211&gt; 162 &lt;212&gt; PRT &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; Synthetic Construct &lt;400&gt; 10

Met Lys lie Leu Lys Pro Tyr Met Arg Asn Thr Ser lie Ser Cys Tyr 15 10 15

Leu Cys Phe Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly He His 20 25 30

Val Phe He Leu Gly Cys Val Ser Val Gly Leu Pro Lys Thr Glu Ala 35 40 45

Asn Trp lie Asp Val Arg Tyr Asp Leu Glu Lys lie Glu Ser Leu lie 50 55 60

Gin Ser He His lie Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His -45 - 96 144 192 240 288 336 384 432 480 489 201207108 80 65 70 75

Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gin 85 90 95

Val lie Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg 100 105 110

Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val 115 120 125

Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe 130 135 140

Thr Glu Phe Leu Gin Ser Phe lie Arg lie Val Gin Met Phe lie Asn 145 150 155 160

Thr Ser &lt;210&gt; 11 &lt;211&gt; 489 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic mIL-15 &lt;220&gt;221&gt; CDS &lt;222&gt; 1)..(489) &lt;400&gt; 11 atg aag ate ctg aag ccc tac atg agg acc age ate age tgt tac 48

Met Lys lie Leu Lys Pro Tyr Met Arg Asn Thr Ser lie Ser Cys Tyr 15 10 15 ctg tgc ttc ctg ctg aac age cac ttc ctg acc gag gcc gga ate cac 96

Leu Cys Phe Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly lie His 20 25 30 gtc ttc ate ctg ggc tgc gtg age gtg ggc ctg ccc aag acc gag gcc 144

Val Phe lie Leu Gly Cys Val Ser Val Gly Leu Pro Lys Thr Glu Ala 35 40 45 aac tgg ate gac gtg agg tac gac ctg gag aag ate gag age ctg ate 192

Asn Trp lie Asp Val Arg Tyr Asp Leu Glu Lys lie Glu Ser Leu lie 50 55 60 cag age ate cac ate gac acc acc ctg tac acc gac age gac ttc cac 240

Gin Ser lie His lie Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His 65 70 75 80 ccc age tgc aag gtg acc gcc atg aac tgc ttc ctg ctg gag ctg caa 288

Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gin 85 90 95 gtg ate ctg cac gag tac age aac atg acc ctg aac gag acc gtg agg 336

Val lie Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg 100 105 110 aac gtg ctg tac ctg get aac age acc ctg age age aac aag aac gtg 384

Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val 115 320 125 gcc gag age ggc tgc aag gag tgt gag gag ctg gag gag aag acc ttc 432

Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe -46 - 480 201207108 130 135 140 acc gag ttc etc cag age ttc ate agg ate gtg cag atg ttc ate aac

Thr Glu Phe Leu Gin Ser Phe lie Arg lie Val Gin Met Phe lie Asn 145 150 155 160 489 acc age tga Thr Ser &lt;210&gt; 12 &lt;211&gt; 162 &lt;212&gt; PRT &lt;213&gt;Artificial sequence&lt;220&gt;;&lt;223&gt; Synthetic Construct &lt;400&gt; 12

Met Lys lie Leu Lys Pro Tyr Met Arg Asn Thr Ser lie Ser Cys Tyr 15 10 15 f

Leu Cys Phe Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly lie His 20 25 30

Val Phe lie Leu Gly Cys Val Ser Val Gly Leu Pro Lys Thr Glu Ala 35 40 45

Asn Trp lie Asp Val Arg Tyr Asp Leu Glu Lys lie Glu Ser Leu lie 50 55 60

Gin Ser lie His lie Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His 65 70 75 80

Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gin 85 90 95

Val lie Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg 100 105 110

Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val 〇 115 120 125

Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe 130 135 140

Thr Glu Phe Leu Gin Ser Phe lie Arg lie Val Gin Met Phe lie Asn 145 150 155 160

Thr Ser &lt;210&gt; 13 &lt;211&gt; 1008 &lt;212&gt; DNA &lt;2]3&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic mIL-12, mp40 &lt;220&gt; -47 - 201207108 &lt;221&gt; CDS &lt;222&gt; (1)..(1008) &lt;400&gt; 13 atg tgc ccc cag aag ctg acc ate age tgg ttc gee ate gtg ctg ctg 48

Met Cys Pro Gin Lys Leu Thr lie Ser Trp Phe Ala lie Val Leu Leu 15 10 15 gtg age ccc ctg atg gee atg tgg gag ctg gag aag gac gtg tac gtg 96

Val Ser Pro Leu Met Ala Met Trp Glu Leu Glu Lys Asp Val Tyr Val 20 25 30 gtg gag gtg gac tgg acc ccc gac gee ccc ggc gag acc gtg aac ctg 144

Val Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu 35 40 45 act tgc gac acc ccc gag gag gac gate acc tgg acc age gac cag 192

Thr Cys Asp Thr Pro Glu Glu Asp Asp lie Thr Trp Thr Ser Asp Gin 50 55 60 aga cac ggc gtc ate ggc age ggc aag acc ctg acc ate acc gtg aag 240

Arg His Gly Val lie Gly Ser Gly Lys Thr Leu Thr lie Thr Val Lys 65 70 75 80 gag ttc ctg gac gee gga cag tac acc tgt cac aag ggc ggc gag acc 288

Glu Phe Leu Asp Ala Gly Gin Tyr Thr Cys His Lys Gly Gly Glu Thr 85 90 95 ctg age cac age cac ctg ttg ctg cac aag aag gag aac ggc ate tgg 336

Leu Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly lie Trp 100 105 110 age acc gag ate ctg aag aac ttc aag aac aag acc ttc ctg aag tgc 384

Ser Thr Glu lie Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys 115 120 125 gag gee ccc aac tac age ggc aga ttc acc tgt age tgg ctg gtg cag 432

Glu Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gin 130 135 140 aga aac atg gac ctg aag ttc aac ate aag age age age age ccc 480

Arg Asn Met Asp Leu Lys Phe Asn lie Lys Ser Ser Ser Ser Ser 145 150 155 160 gac age aga gee gtg aca tgc ggc atg gee age ctg age gee gag aag 528

Asp Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys 165 170 175 gtg acc ctg gac cag aga gac tac gag aag tac age gtg age tgc cag 576

Val Thr Leu Asp Gin Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gin 180 185 190 gag gac gtg acc tgt ccc acc gee gag gag acc ctg ccc ate gag ett 624

Glu Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro lie Glu Leu 195 200 205 gee ctg gaa gee aga cag cag aac aag tac gag aac tac age acc age 672

Ala Leu Glu Ala Arg Gin Gin Asn Lys Tyr Glu Asn Tyr Ser Thr Ser 210 215 220 ttc ttc ate aga gac ate ate aag ccc gac ccc ccc aag aac etc cag 720

Phe Phe lie Arg Asp lie He Lys Pro Asp Pro Pro Lys Asn Leu Gin 225 230 235 240 atg aag ccc ctg aag aac age cag gtg gag gtg tee tgg gag tac ccc 768

Met Lys Pro Leu Lys Asn Ser Gin Val Glu Val Ser Trp Glu Tyr Pro 245 250 255 gac age tgg age acc ccc cac age tac ttc age ctg aag ttc ttc gtg 816

Asp Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val 260 265 270 aga ate cag aga aag aag gag aag atg aag gag acc gag gag ggc tgc 864

Arg lie Gin Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys 275 280 285 -48 - 912 201207108 aac cag aag ggc get ttc ctg gtg gag aaa acc age acc gag gtg cag

Asn Gin Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gin 290 295 300 tgc aag ggc ggc aac gtg tgt gtg cag gee cag gac aga tac tac aac

Cys Lys Gly Gly Asn Val Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn 305 310 315 320 age age tgc tee aag tgg gee tgc gtg ccc tgc ege gtg aga age tga Ser Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser 325 330 335 &lt;210> 14 &lt;211&gt; 335 &lt;212&gt; PRT &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic construct &lt;400&gt; 14 c

960 1008

Met Cys Pro Gin Lys Leu Thr lie Ser Trp Phe Ala lie Val Leu Leu 15 10 15

Val Ser Pro Leu Met Ala Met Trp Glu Leu Glu Lys Asp Val Tyr Val 20 25 30

Val Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu 35 40 45

Thr Cys Asp Thr Pro Glu Glu Asp Asp lie Thr Trp Thr Ser Asp Gin 50 55 60

Arg His Gly Val lie Gly Ser Gly Lys Thr Leu Thr lie Thr Val Lys 65 70 75 80

Glu Phe Leu Asp Ala Gly Gin Tyr Thr Cys His Lys Gly Gly Glu Thr 85 90 95

Leu Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly lie Trp 100 105 110

Ser Thr Glu lie Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys 115 120 125

Glu Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gin 130 335 140

Arg Asn Met Asp Leu Lys Phe Asn lie Lys Ser Ser Ser Ser Ser 145 150 155 160

Asp Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys 165 170 175

Val Thr Leu Asp Gin Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gin 180 185 190

Glu Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro lie Glu Leu 195 200 205 .49 - 201207108

Ala Leu Glu Ala Arg Gin Gin Asn Lys Tyr Glu Asn Tyr Ser Thr Ser 210 215 220

Phe Phe lie Arg Asp lie lie Lys Pro Asp Pro Pro Lys Asn Leu Gin 225 230 235 240

Met Lys Pro Leu Lys Asn Ser Gin Val Glu Val Ser Trp Glu Tyr Pro 245 250 255

Asp Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val 260 265 270

Arg lie Gin Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys 275 280 285

Asn Gin Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gin 290 295 300

Cys Lys Gly Gly Asn Val Cys Val Gin Ala Gin Asp Arg Tyr Tyr Asn 305 310 315 320

Ser Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser 325 330 335 &lt;210&gt; 15 &lt;211&gt; 648 &lt;212&gt; DNA &lt;213&gt;Artificial Sequence&lt;220&gt;&lt;223&gt; Synthetic mIL -12, mp35 &lt;220〉 &lt;221&gt; CDS &lt;222&gt; (1)..(648) &lt;400&gt; 15 atg tgc cag age aga tac ctg ttg ttc ctg get acc ctg gcc ctg ctg 48

Met Cys Gin Ser Arg Tyr Leu Leu Phe Leu Ala Thr Leu Ala Leu Leu 15 10 15 aac cac ctg age ctg gcc ege gtg ate ccc gtg age ggc ccc gcc aga 96

Asn His Leu Ser Leu Ala Arg Val lie Pro Val Ser Gly Pro Ala Arg 20 25 30 tgc ctg age cag age aga aac ctg ttg aaa aca acc gac gac atg gtg 144

Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val 35 40 45 aaa acc gcc aga gag aag ctg aag cac tac age tgc acc gcc gag gac 192

Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp 50 55 60 ate gac cac gag gac ate acc aga gac cag acc age acc ctg aaa acc 240 lie Asp His Glu Asp lie Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr 65 70 75 80 tgt ctg ccc ctg gag ctg cac aag aac gag age tgc ctg get acc aga 288

Cvs Leu Pr〇 Leu Glu Len His hn G】u Ser Cvs A]a Thr Arg J 85 90 95 gag acc age age acc acc aga ggc age tgc ctg ccc ccc cag aaa acc 336

Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr 100 105 110 -50 - 384 201207108 age ctg atg atg acc ctg tgc ctg ggc age ate tac gag gac ctg aag

Ser Leu Met Met Thr Leu Cys Leu Gly Ser lie Tyr Glu Asp Leu Lys 115 120 125 atg tac cag acc gag ttc cag gcc ate aac gcc gcc ctg caa aac cac

Met Tyr Gin Thr Glu Phe Gin Ala lie Asn Ala Ala Leu Gin Asn His 130 135 140 aac cac cag cag ate ate ctg gac aag ggc atg ttg gtg gcc ate gac

Asn His Gin Gin lie lie Leu Asp Lys Gly Met Leu Val Ala lie Asp 145 150 155 160 gag ctg atg cag age ctg aac cac aac ggc gag acc ctg aga cag aag

Glu Leu Met G]n Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gin Lys 165 170 175 ccc ccc gtg ggc gag gcc gac ccc tac aga gtg aag atg aag ctg tgc

Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys 180 185 190 ate ctg ctg cac gcc ttc age acc aga gtg lie Leu Leu His Ala Phe Ser Thr Arg Val Val Thr lie Asn Arg Val 195 200 205

432 480 528 576 624 648 atg ggc tac ctg age age gcc tga Met Gly Tyr Leu Ser Ser Ala 210 215 &lt;210&gt; 16 &lt;211&gt; 215 &lt;212&gt; PRT &lt;213&gt;Artificial sequence &lt;220&gt;223&gt; Synthetic Construct &lt;400&gt; 16

Met Cys Gin Ser Arg Tyr Leu Leu Phe Leu Ala Thr Leu Ala Leu Leu 15 10 15

Asn His Leu Ser Leu Ala Arg Val lie Pro Val Ser Gly Pro Ala Arg 20 25 30

Cys Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val 35 40 45

Lys Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp 50 55 60 lie Asp His Glu Asp lie Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr 65 70 75 80

Cys Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg 85 90 95

Glu Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr 100 105 110

Ser Leu Met Met Thr Leu Cys Leu Gly Ser lie Tyr Glu Asp Leu Lys 115 120 125

Met Tyr Gin Thr Glu Phe Gin Ala lie Asn Ala Ala Leu Gin Asn His 130 135 140 -51 - 201207108

Asn His Gin Gin lie lie Leu Asp Lys Gly Met Leu Val Ala lie Asp 145 150 155 160

Glu Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gin Lys 165 170 175

Pro Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys 180 185 190 lie Leu Leu His Ala Phe Ser Thr Arg Val Val Thr lie Asn Arg Val 195 200 205

Met Gly Tyr Leu Ser Ser Ala 210 215 &lt;210&gt; 17 &lt;211&gt; 489 &lt;212&gt; DNA &lt;213&gt;Artificial Sequence&lt;220&gt;&lt;223&gt; Synthetic hIL-21 &lt;220&gt;&lt;223&gt; CDS &lt;222&gt; (1)..(489) &lt;400&gt; 17 atg aga age age ccc ggc aac atg gag aga ate gtg ate tgc ctg atg 48

Met Arg Ser Ser Pro Gly Asn Met Glu Arg lie Val lie Cys Leu Met 15 10 15 gtg ate ttc ctg ggc acc ctg gtg cat aag age age age cag ggc cag 96

Val lie Phe Leu Gly Thr Leu Val His Lys Ser Ser Ser Gin Gly Gin 20 25 30 gac aga cac atg ate ege atg aga cag ctg ate gac ate gtg gac cag 144

Asp Arg His Met lie Arg Met Arg Gin Leu lie Asp lie Val Asp Gin 35 40 45 ctg aag aac tac gtg aac gac ctg gtg ccc gag ttc ctg ccc gcc ccc 192

Leu Lys Asn Tyr Val Asn Asp Leu Val Pro Glu Phe Leu Pro Ala Pro 50 55 60 gag gac gtg gag acc aac tgc gag tgg age gcc ttc age tgc ttc cag 240

Glu Asp Val Glu Thr Asn Cys Glu Trp Ser Ala Phe Ser Cys Phe Gin 65 70 75 80 aag gcc cag ctg aag tcc gcc aac acc ggc aac aac gag aga ate ate 288

Lys Ala Gin Leu Lys Ser Ala Asn Thr Gly Asn Asn Glu Arg lie lie 85 90 95 aac gtg age ate aag aag ctg aag egg aag ccc ccc age acc aac gcc 336

Asn Val Ser lie Lys Lys Leu Lys Arg Lys Pro Pro Ser Thr Asn Ala 100 105 110 gga aga aga cag aag cac aga ctg acc tgt ccc age tgc gac age tac 384

Gly Arg Arg Gin Lys His Arg Leu Thr Cys Pro Ser Cys Asp Ser Tyr 115 120 125 gag aag aag ccc ccc aag gag ttc ctg gag aga ttc aag age ctg ctg 432

Glu Lys Lys Pro Pro Lys Glu Phe Leu Glu Arg Phe Lys Ser Leu Leu 130 135 140 caa aag at ate cac cag cac ctg age age aga acc cac ggc age gag 480

Gin Lys Met lie His Gin His Leu Ser Ser Arg Thr His Gly Ser Glu 145 150 155 160 gac age tga 489 -52 - 201207108

Asp Ser &lt;210> 18 &lt;211&gt; 162 &lt;212&gt; PRT &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic construct &lt;400&gt;

Met Arg Ser Ser Pro Gly Asn Met Glu Arg lie Val lie Cys Leu Met 1 5 10 15

Val lie Phe Leu Gly Thr Leu Val His Lys Ser Ser Ser Gin Gly Gin 20 25 30

Asp Arg His Met lie Arg Met Arg Gin Leu lie Asp lie Val Asp Gin 35 40 45 f

Leu Lys Asn Tyr Val Asn Asp Leu Val Pro Glu Phe Leu Pro Ala Pro 50 55 60

Glu Asp Val Glu Thr Asn Cys Glu Trp Ser Ala Phe Ser Cys Phe Gin 65 70 75 80

Lys Ala Gin Leu Lys Ser Ala Asn Thr Gly Asn Asn Glu Arg lie lie 85 90 95

Asn Val Ser lie Lys Lys Leu Lys Arg Lys Pro Pro Ser Thr Asn Ala 100 105 110

Gly Arg Arg Gin Lys His Arg Leu Thr Cys Pro Ser Cys Asp Ser Tyr 115 120 125

Glu Lys Lys Pro Pro Lys Glu Phe Leu Glu Arg Phe Lys Ser Leu Leu 130 135 140

Gin Lys Met lie His Gin His Leu Ser Ser Arg Thr His Gly Ser Glu 145 150 155 160

Asp Ser &lt;210&gt; 19 &lt;211&gt; 489 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic hIL-15 &lt;220&gt;&lt;221&gt; CDS &lt;222&gt; 1)..(489) &lt;400&gt; 19 atg aga ate age aag ccc cac ctg aga age ate age ate cag tgt tac 48

Met Arg lie Ser Lys Pro His Leu Arg Ser lie Ser lie Gin Cys Tyr 15 10 15 -53 - 201207108 ctg tgc ctg ctg ctg aac age cac ttc ctg acc gag gee ggt ate cac 96

Leu Cys Leu Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly lie His 20 25 30 gtc ttc ate ctg ggc tgc ttc age gee gga ctg ccc aag acc gag gee 144

Val Phe He Leu Gly Cys Phe Ser Ala Gly Leu Pro Lys Thr Glu Ala 35 40 45 aac tgg gtg aac gtg ate tet gac ctg aag aag ate gag gac ctg ate 192

Asn Trp Val Asn Val lie Ser Asp Leu Lys Lys lie Glu Asp Leu lie 50 55 60 cag tee atg cac ate gac gee acc ctg tac acc gag age gac gtt cat 240

Gin Ser Met His lie Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His 65 70 75 80 ccc age tgc aag gtg acc gee atg aag tgc ttc ctg ctg gag ctg caa 288

Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gin 85 90 95 gtg ate tee ctg gag age ggc gac gee age ate cac gac acc gtg gag 336

Val lie Ser Leu Glu Ser Gly Asp Ala Ser lie His Asp Thr Val Glu 100 105 110 aac ctg att ate ctg get aac aac age ctg age age aac ggc aac gtg 384

Asn Leu lie lie Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val 115 120 125 acc gag age ggc tgc aag gag tgt gag gag ctg gag gag aag aac ate 432

Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn lie 130 135 140 aag gag ttc etc cag age ttc gtg cat ate gtc cag atg ttc ate aac 480

Lys Glu Phe Leu Gin Ser Phe Val His lie Val Gin Met Phe lie Asn 145 150 155 160 acc age tga 489

Thr Ser &lt;210&gt; 20 &lt;211> 162 &lt;212&gt; PRT &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Synthetic Construct &lt;400&gt; 20

Met Arg lie Ser Lys Pro His Leu Arg Ser lie Ser lie Gin Cys Tyr 15 10 15

Leu Cys Leu Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly lie His 20 25 30

Val Phe lie Leu Gly Cys Phe Ser Ala Gly Leu Pro Lys Thr Glu Ala 35 40 45

Asn Trp Val Asn Val lie Ser Asp Leu Lys Lys lie Glu Asp Leu lie 50 55 60

Gin Ser Met His lie Aso Ala Thr Leu Tvr Thr Glu Ser Asd Val His 65 70' 75 ^ 80

Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gin 85 90 95 -54 -

c

201207108

Val lie Ser Leu Glu Ser Gly Asp Ala Ser lie His Asp Thr Val Glu 100 105 110

Asn Leu lie lie Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val 115 120 125

Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn lie 130 135 140

Lys Glu Phe Leu Gin Ser Phe Val His lie Val Gin Met Phe lie Asn 145 150 155 160

Thr Ser &lt;210> 21 &lt;211&gt; 987 &lt;212&gt; DNA &lt;213&gt;Artificial Sequence&lt;220&gt;&lt;223&gt; Synthetic hIL-12, p40 &lt;220&gt;&lt;221&gt; CDS &lt;222&gt;; (1)..(987) &lt;400&gt; 21 atg ggt cac cag cag ttg gtc ate tet tgg ttt tcc ctg gtt ttt ctg

Met Gly His Gin Gin Leu Val lie Ser Trp Phe Ser Leu Val Phe Leu 15 10 15 gca tet ccc etc gtg gcc ata tgg gaa ctg aag aaa gat gtt tat gtc

Ala Ser Pro Leu Val Ala lie Trp Glu Leu Lys Lys Asp Val Tyr Val 20 25 30 gta gaa ttg gat tgg tat ccg gat gcc cct gga gaa atg gtg gtc etc

Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met Val Val Leu 35 40 45 acc tgt gac acc cct gaa gaa gat ggt ate acc tgg acc ttg gac cag

Thr Cys Asp Thr Pro Glu Glu Asp Gly lie Thr Trp Thr Leu Asp Gin 50 55 60 age agt gag gtc tta ggc tet ggc aaa acc ctg acc ate caa gtc aaa

Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr lie Gin Val Lys 65 70 75 80 gag ttt gga gat get ggc cag tac acc tgt cac aaa gga ggc gag gtt Glu Phe Gly Asp Ala Gly Gin Tyr Thr Cys His Lys Gly Gly Glu Val 85 90 95 eta age cat teg etc ctg ctg ett cac aaa aag gaa gat gga att tgg Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp Gly lie Trp 100 105 110 tcc act gat att tta aag gac cag aaa gaa Ccc aaa aat aag acc ttt Ser Thr Asp lie Leu Lys Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe 115 120 125 eta aga tgc gag gcc aag aat tat tet gga cgt ttc acc tgc tgg tgg

Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp 130 135 140 ctg aeg aca ate agt act gat ttg aca ttc agt gtc aaa age age aga

Leu Thr Thr lie Ser Thr Asp Leu Thr Phe Ser Val Lys Ser Ser Arg 145 150 155 160 48 96 144 192 240 288 336 384 432 -55 - 480 528 201207108 ggc tct tct gac ccc caa ggg gtg acg tgc gga get get aca etc Tet

Gly Ser Ser Asp Pro G】n G】y Va】Thr Cys Gly A]a Ala Thr Leu Ser 165 170 175 gca gag aga gtc aga ggg gac aac aag gag tat gag tac tea gtg gag

Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu 180 185 190 tgc cag gag gac agt gee tgc cca get get gag gag agt ctg ccc att

Cys Gin Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser Leu Pro lie 195 200 205 gag gtc atg gtg gat gee gtt cac aag etc aag tat gaa aac tac acc

Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu Asn Tyr Thr 210 215 220 age age ttc ttc ate agg gac ate ate aaa cct gac cca ccc aag aac

Ser Ser Phe lie Arg Asp lie lie Lys Pro Asp Pro Pro Lys Asn 225 230 235 240 ttg cag ctg aag cca tta aag aat tct egg cag gtg gag gtc age tgg

Leu Gin Leu Lys Pro Leu Lys Asn Ser Arg Gin Val Glu Val Ser Trp 245 250 255 gag tac cct gac acc tgg agt act cca cat tee tac ttc tee ctg aca

Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Thr 260 265 270 ttc tgc gtt cag gtc cag ggc aag age aag aga gaa aag aaa gat aga

Phe Cys Val Gin Val Gin Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg 275 280 285 gtc ttc acg gac aag acc tea gee acg gtc ate tgc ege aaa aat gee

Val Phe Thr Asp Lys Thr Ser Ala Thr Val He Cys Arg Lys Asn Ala 290 295 300 age att gtg egg gee cag gac ege tac tat age tea tct tgg age

Ser lie Ser Val Arg Ala Gin Asp Arg Tyr Tyr Ser Ser Ser Trp Ser 305 310 315 320 gaa tgg gca tct gtg ccc tgc agt tag

Glu Trp Ala Ser Val Pro Cys Ser 325 &lt;210> 22 &lt;211&gt; 328 &lt;212&gt; PRT &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic construct &lt;400&gt;

Met Gly His Gin Gin Leu Val lie Ser Trp Phe Ser Leu Val Phe Leu 15 10 15

Ala Ser Pro Leu Val Ala lie Trp Glu Leu Lys Lys Asp Val Tyr Val 20 25 30

Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met Val Val Leu 35 40 45

Thr Cys Asp Thr Pro Glu Glu Asp Gly He Thr Trp Thr Leu Asp Gin 50 55 60

Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr lie Gin Val Lys 65 70 75 80 576 624 672 720 768 816 864 912 960 987 -56 - 201207108

Glu Phe Gly Asp Ala Gly Gin Tyr Thr Cys His Lys Gly Gly Glu Val 85 90 95

Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp Gly lie Trp 100 105 110

Ser Thr Asp lie Leu Lys Asp Gin Lys Glu Pro Lys Asn Lys Thr Phe 115 120 125

Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp 130 135 140

Leu Thr Thr lie Ser Thr Asp Leu Thr Phe Ser Val Lys Ser Ser Arg 145 150 155 160

Gly Ser Ser Asp Pro Gin Gly Val Thr Cys Gly Ala Ala Thr Leu Ser 165 170 175

Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu 180 185 190

Cys Gin Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser Leu Pro lie 195 200 205

Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu Asn Tyr Thr 210 215 220

Ser Ser Phe lie Arg Asp lie lie Lys Pro Asp Pro Pro Lys Asn 225 230 235 240

Leu Gin Leu Lys Pro Leu Lys Asn Ser Arg Gin Val Glu Val Ser Trp 245 250 255

Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Thr 260 265 270

Phe Cys Val Gin Val Gin Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg 275 280 285

Val Phe Thr Asp Lys Thr Ser Ala Thr Val lie Cys Arg Lys Asn Ala 290 295 300

Ser lie Ser Val Arg Ala Gin Asp Arg Tyr Tyr Ser Ser Ser Trp Ser 305 310 315 320

Glu Trp Ala Ser Val Pro Cys Ser 325 &lt;210&gt; 23 &lt;211&gt; 660 &lt;212&gt; DNA &lt;213&gt;Artificial Sequence &lt;220&gt;&lt;223&gt; Synthetic hIL-12, ρ35

&lt;220&gt;&lt;221&gt; CDS • 57 - 201207108 &lt;222&gt; (1)..(660) &lt;400&gt; 23 atg ggt cca gcg cgc age etc etc ett gtg get acc ctg gtc etc ctg 48

Met Gly Pro Ala Arg Ser Leu Leu Leu Val Ala Thr Leu Val Leu Leu 15 10 15 gac cac etc agt ttg gee aga aac etc ccc gtg gee act cca gac cca 96

Asp His Leu Ser Leu Ala Arg Asn Leu Pro Val Ala Thr Pro Asp Pro 20 25 30 gga atg ttc cca tgc ett cac cac tee caa aac ctg ctg agg gee gtc 144

Gly Met Phe Pro Cys Leu His His Ser Gin Asn Leu Leu Arg Ala Val 35 40 45 age aac atg etc cag aag gee aga caa act eta gaa ttt tac cct tgc 192

Ser Asn Met Leu Gin Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys 50 55 60 act tet gaa gag att gat cat gaa gat ate aca aaa gat aaa acc age 240

Thr Ser Glu Glu lie Asp His Glu Asp lie Thr Lys Asp Lys Thr Ser 65 70 75 80 aca gtg gag gee tgt tta cca ttg gaa tta acc aag aat gag agt tgc 288

VAL aat tee aga gag acc tet ttc ata act aat ggg agt tgc ctg gee 336

Leu Asn Ser Arg Glu Thr Ser Phe lie Thr Asn Gly Ser Cys Leu Ala 100 105 110 tee aga aag acc tet ttt atg atg gee ctg tgc ett agt agt att tat 384

Ser Arg Lys Thr Ser Phe Met Met Ala Leu Cys Leu Ser Ser lie Tyr 115 120 125 gaa gac ttg aag atg tac cag gtg gag ttc aag acc atg aat gca aag 432

Glu Asp Leu Lys Met Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys 130 135 140 ett ctg atg gat cct aag agg cag ate ttt eta gat caa aac atg ctg 480

Leu Leu Met Asp Pro Lys Arg Gin lie Phe Leu Asp Gin Asn Met Leu 145 150 155 160 gca gtt att gat gag ctg atg cag gee ctg aat ttc aac agt gag act 528

Ala Val lie Asp Glu Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr 165 170 175 gtg cca caa aaa tee tee ett gaa gaa ccg gat ttt tat aaa act aaa 576

Val Pro Gin Lys Ser Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys 180 185 190 ate aag etc tgc ata ett ett cat get ttc aga att egg gca gtg act 624 lie Lys Leu Cys lie Leu Leu His Ala Phe Arg lie Arg Ala Val Thr 195 200 205 att gat aga gtg atg age tat ctg aat get tee taa 660 lie Asp Arg Val Met Ser Tyr Leu Asn Ala Ser 210 215 &lt;210&gt; 24 &lt;211&gt; 219 &lt;212&gt; PRT &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Synthetic Construct &lt;400&gt; 24

Met Gly Pro Ala Arg Ser Leu Leu Leu Val Ala Thr Leu Val Leu Leu 15 10 15

Asp His Leu Ser Leu Ala Arg Asn Leu Pro Val Ala Thr Pro Asp Pro -58 - 201207108 20 25 30

Gly Met Phe Pro Cys Leu His His Ser Gin Asn Leu Leu Arg Ala Val 35 40 45

Ser Asn Met Leu Gin Lys Ala Arg Gin Thr Leu Glu Phe Tyr Pro Cys 50 55 60

Thr Ser Glu Glu lie Asp His Glu Asp lie Thr Lys Asp Lys Thr Ser 65 70 75 80

Thr Val Glu Ala Cys Leu Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys 85 90 95

Leu Asn Ser Arg Glu Thr Ser Phe lie Thr Asn Gly Ser Cys Leu Ala 100 105 110

Ser Arg Lys Thr Ser Phe Met Met Ala Leu Cys Leu Ser Ser lie Tyr 115 120 125

Glu Asp Leu Lys Met Tyr Gin Val Glu Phe Lys Thr Met Asn Ala Lys 130 135 140

Leu Leu Met Asp Pro Lys Arg Gin lie Phe Leu Asp Gin Asn Met Leu 145 150 155 160

Ala Val lie Asp Glu Leu Met Gin Ala Leu Asn Phe Asn Ser Glu Thr 165 170 175

Val Pro Gin Lys Ser Ser Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys 180 185 190 lie Lys Leu Cys lie Leu Leu His Ala Phe Arg lie Arg Ala Val Thr 195 200 205 lie Asp Arg Val Met Ser Tyr Leu Asn Ala Ser 210 215

&lt;210&gt; 25 &lt;211&gt; 17 &lt;212&gt; PRT &lt;213&gt; Drosophila melanogaster &lt;400&gt; 25

Arg Arg Gly Gly Thr Thr Cys Ala Asn Thr Gly Ala Cys Ala Cys Tyr 15 10 15

Tyr

&lt;210&gt; 26 &lt;211&gt; 13 &lt;212&gt; DNA &lt;213&gt; Drosophila melanogaster &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (7)..(7) -59 - 201207108 &lt;223&gt; n is a, c, g, or t &lt;400&gt; 26 aggtcanagg tea &lt;210&gt; 27 &lt;211&gt; 15 &lt;212&gt; DNA &lt;213&gt; Drosophila melanogaster &lt;400&gt; 27 gggttgaatg aattt 15 &lt;210&gt; 28 &lt;211&gt; 18 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic homing nucleic acid? (HE) (I-Scel) &lt;400&gt; 28 tagggataac agggtaat 18 &lt;210> 29 &lt;211&gt; 37323 &lt;212&gt; DNA &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; Synthetic Ad-RTS -hIL-12 (SPl-RheoIL-12) &lt; 400 &gt; 29 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa gegaeggatg tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca tagegegtaa tatttgtcta gggagatccg gtaccggcgc gcgcgccgtt tggccgcctc gagtetagag atccggtgag tattaggege gcaccaggtg ccgcaataaa atatetttat tttcattaca tctgtgtgtt ggttttttgt gtgaategat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttetetat egataatgea ggtcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg aga ctcttcg aaggaagagg ggcggggtcg atcgaccccg cccctcttcc ttcgaaggaa gaggggcggg gtcgaagacc tagagggtat ataatgggtg ccttagctgg tgtgtgagct catcttcctg tagatcacgc gtgccaccat gggtcaccag cagttggtca tctcttggtt ttccctggtt tttctggcat ctcccctcgt ggccatatgg gaactgaaga aagatgttta tgtcgtagaa ttggattggt atccggatgc ccctggagaa atggtggtcc tcacctgtga cacccctgaa gaagatggta tcacctggac cttggaccag ageagtgagg tcttaggctc tggcaaaacc ctgaccatcc aagtcaaaga gtttggagat gctggccagt acacctgtca caaaggaggc gaggttctaa gccattcgct cctgctgctt cacaaaaagg aagatggaat -60 - 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 Π40 1200

201207108 ttggtccact gatattttaa aggaccagaa agaacccaaa aataagacct ttctaagatg cgaggccaag aattattctg gacgtttcac ctgctggtgg ctgacgacaa tcagtactga tttgacattc agtgtcaaaa gcagcagagg ctcttctgac ccccaagggg tgacgtgcgg agctgctaca ctctctgcag agagagtcag aggggacaac aaggagtatg agtactcagt ggagtgccag gaggacagtg cctgcccagc tgctgaggag agtctgccca ttgaggtcat ggtggatgcc gttcacaagc tcaagtatga aaactacacc agcagcttct tcatcaggga catcatcaaa cctgacccac ccaagaactt gcagctgaag ccattaaaga attctcggca ggtggaggtc agctgggagt accctgacac ctggagtact ccacattcct acttctccct gacattctgc gttcaggtcc agggcaagag caagagagaa aagaaagata gagtcttcac ggacaagacc tcagccacgg tcatctgccg caaaaatgcc agcattagcg tgcgggccca ggaccgctac tatagctcat cttggagcga atgggcatct gtgccctgca gttaggttgg gcgagctcga attcattgat cccccgggct gcaggaattc gatatcaagc tcgggatccg aattccgccc cccccccccc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgggt ccagcgcgca gcctcctcct tgtggctacc ctggtcctcc tggaccacct cagtttggcc agaaacctcc ccgtggccac tccagaccca ggaatgttcc catgccttca ccactcccaa aacctgctga gggccgtcag caacatgctc cagaaggcca gacaaactct agaattttac ccttgcactt ctgaagagat tgatcatgaa gatatcacaa aagataaaac cagcacagtg gaggcctgtt taccattgga attaaccaag aatgagagtt gcctaaattc cagagagacc tctttcataa ctaatgggag ttgcctggcc tccagaaaga cctcttttat gatggccctg tgccttagta gtatttatga agacttgaag atgtaccagg tggagttcaa gaccatgaat gcaaagcttc tgatggatcc taagaggcag atctttctag atcaaaacat gctggcagtt attgatgagc tgatgcaggc cctgaatttc aacagtgaga ctgtgccaca aaaatcctcc cttgaagaac cggattttta taaaactaaa atcaagctct gcatacttct tcatgctttc agaattcggg cagtgactat tgatagagtg atgagctatc tgaatgcttc ctaacgtacg tcgacatcga gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgggcgcgcc ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 -61 - 201207108 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag 3480 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg 3540 ttt tctttcc agagagcgga acaggcgagg Aaaagtagtc ccttctcggc gattctgcgg 3600 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac 3660 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgc tgtgatc 3720 gtcacttggt gagtagcggg ctgctgggct gggtacgtgc gctcggggtt ggcgagtgtg 3780 ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt 3840 cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac 3900 gagctagcgc cgccaccatg ggccctaaaa agaagcgtaa agtcgccccc ccgaccgatg 3960 tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg catgccgacg 4020 cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt ccgggattta 4080 ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag tttgagcaga 4140 tgtttaccga tgcccttgga attgacgagt acggtgggga attcgagatg cctgtggaca 4200 ggatcctgga ggcagagctt gctgtggaac agaagagtga ccagggcgtt gagggtcctg 4260 ggggaaccgg gggtagcggc agcagcccaa atgaccctgt gactaacatc tgtcaggcag 4320 ctgacaaaca gctattcacg cttgttgagt gggcgaagag gatcccacac ttttcctcct 4380 tgcctctgga tgatcaggtc atattgctgc gggcaggctg gaatgaactc ctcattgcct 4440 cct 11 tcaca ccgatccatt gatgttcgag atggcatcct ccttgccaca ggtcttcacg 4500 tgcaccgcaa ctcagcccat tcagcaggag taggagccat cttt gatcgg gtgctgacag 4560 agctagtgtc caaaatgcgt gacatgagga tggacaagac agagcttggc tgcctgaggg 4620 caatcattct gtttaatcca gaggtgaggg gtttgaaatc cgcccaggaa gttgaacttc 4680 tacgtgaaaa agtatatgcc gctttggaag aatatactag aacaacacat cccgatgaac 4740 caggaagatt tgcaaaactt ttgcttcgtc tgccttcttt acgttccata ggccttaagt 4800 gtttggagca tttgtttttc tttcgcctta ttggagatgt tccaattgat acgttcctga 4860 tggagatgct tgaatcacct tctgattcat aatctagcct agcccccctc tccctccccc 4920 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg 4980 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc 5040 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg 5100 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg 5160 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca 5220 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata 5280 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc 5340 cagaaggtac cccattgtat gggatctgat ctggggc ctc ggtgcacatg ctttacatgt 5400 gtttagtcga ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt 5460 gaaaaacacg atctctaggc gccaccatga agctactgtc ttctatcgaa caagcatgcg 5520 atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc 5580 tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg ctgactaggg 5640 cacatctgac agaagtggaa tcaaggctag aaagactgga acagctattt -62 - ctactgattt 5700 201207108

ttcctcgaga agaccttgac atgattttga aaatggattc tttacaggat ataaaagcat 5760 tgttaacagg attatttgta caagataatg tgaataaaga tgccgtcaca gatagattgg 5820 cttcagtgga gactgatatg cctctaacat tgagacagca tagaataagt gcgacatcat 5880 catcggaaga gagtagtaac aaaggtcaaa gacagttgac tgtatcgccg gaattcccgg 5940 ggatccggcc tgagtgcgta gtacccgaga ctcagtgcgc catgaagcgg aaagagaaga 6000 aagcacagaa ggagaaggac aaactgcctg tcagcacgac gacggtggac gaccacatgc 6060 cgcccattat gcagtgtgaa cctccacctc ctgaagcagc aaggattcac gaagtggtcc 6120 caaggtttct ctccgacaag ctgttggtga caaaccggca gaaaaacatc ccccagttga 6180 cagccaacca gcagttcctt atcgccaggc tcatctggta ccaggacggg tacgagcagc 6240 cttctgatga agatttgaag aggattacgc agacgtggca gcaagcggac gatgaaaacg 6300 aagagtcgga cactccct tc cgccagatca cagagatgac tatcctcacg gtccaactta 6360 tcgtggagtt cgcgaaggga ttgccagggt tcgccaagat ctcgcagcct gatcaaatta 6420 cgctgcttaa ggcttgctca agtgaggtaa tgatgctccg agtcgcgcga cgatacgatg 6480 cggcctcaga cagtattctg ttcgcgaaca accaagcgta cactcgcgac aactaccg ca 6540 aggctggcat ggccgaggic atcgaggatc tactgcactt ctgccggtgc atgtactcta 6600 tggcgttgga caacatccat tacgcgctgc tcacggctgt cgtcatcttt tctgaccggc 6660 cagggttgga gcagccgcaa ctggtggaag agatccagcg gtactacctg aatacgctcc 6720 gcatctatat cctgaaccag ctgagcgggt cggcgcgttc gtccgtcata tacggcaaga 6780 tcctctcaat cctctctgag ctacgcacgc tcggcatgca aaactccaac atgtgcatct 6840 ccctcaagct caagaacaga aagctgccgc ctttcctcga ggagatctgg gatgtggcgg 6900 acatgtcgca cacccaaccg ccgcctatcc tcgagtcccc cacgaatctc taggcggcct 6960 ctagagcggc cgccaccgcg gggagatcca gacatgataa gatacattga tgagtttgga 7020 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt 7080 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat 7140 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac 7200 aaatgtggta tggctgatta tgatccggct gcctcgcgcg tttcggtgat gacggtgaaa 7260 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 7320 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga 7380 ggtcgactct agtccccgcg gtggcagatc tggaaggtgc tgaggtacga tgagacccgc 7440 accaggtgca gaccctgcga gtgtggcggt aaacatatta ggaaccagcc tgtgatgctg 7500 gatgtgaccg aggagctgag gcccgatcac ttggtgctgg cctgcacccg cgctgagttt 7560 ggctctagcg atgaagatac agattgaggt actgaaatgt gtgggcgtgg cttaagggtg 7620 ggaaagaata tataaggtgg gggtcttatg tagttttgta tctgttttgc agcagccgcc 7680 gccgccatga gcaccaactc gtttgatgga agcattgtga gctcatattt gacaacgcgc 7740 atgcccccat gggccggggt gcgtcagaat gtgatgggct ccagcattga tggtcgcccc 7800 gtcctgcccg caaactctac taccttgacc tacgagaccg tgtctggaac gccgttggag 7860 actgcagcct ccgccgccgc ttcagccgct gcagccaccg cccgcgggat tgtgactgac 7920 -63 · 201207108 tttgctttcc tgagcccgct tgcaagcagt gcagcttccc gttcatccgc ccgcgatgac 7980 aagttgacgg ctcttttggc acaattggat tctttgaccc gggaacttaa tgtcgtttct 8040 cagcagctgt tggatctgcg ccagcaggtt tctgccctga aggcttcctc ccctcccaat 8100 gcggtttaaa acataaataa aaaaccagac tctgtttgga tttggatcaa gcaagtgtct 8160 tgctgtcttt atttaggggt tttgcgcgc g cggtaggccc gggaccagcg gtctcggtcg 8220 ttgagggtcc tgtgtatttt ttccaggacg tggtaaaggt gactctggat gttcagatac 8280 atgggcataa gcccgtctct ggggtggagg tagcaccact gcagagcttc atgctgcggg 8340 gtggtgttgt agatgatcca gtcgtagcag gagcgctggg cgtggtgcct aaaaatgtct 8400 ttcagtagca agctgattgc caggggcagg cccttggtgt aagtgtttac aaagcggtta 8460 agctgggatg ggtgcatacg tggggatatg agatgcatct tggactgtat ttttaggttg 8520 gctatgttcc cagccatatc cctccgggga ttcatgttgt gcagaaccac cagcacagtg 8580 tatccggtgc acttgggaaa tttgtcatgt agcttagaag gaaatgcgtg gaagaacttg 8640 gagacgccct tgtgacctcc aagattttcc atgcattcgt ccataatgat ggcaatgggc 8700 ccacgggcgg cggcctgggc gaagatattt ctgggatcac taacgtcata gttgtgttcc 8760 aggatgagat cgtcataggc cattt ttaca aagcgcgggc ggagggtgcc agactgcggt 8820 ataatggttc catccggccc aggggcgtag ttaccctcac agatttgcat ttcccacgct 8880 ttgagttcag atggggggat catgtctacc tgcggggcga tgaagaaaac ggtttccggg 8940 gtaggggaga tcagctggga agaaagcagg ttcctgagca gctgcgactt accgcagccg 9000 gtgggcccgt aaatcacacc tattaccggc tgcaactggt agttaagaga gctgcagctg 9060 ccgtcatccc tgagcagggg ggccacttcg ttaagcatgt ccctgactcg catgttttcc 9120 ctgaccaaat ccgccagaag gcgctcgccg cccagcgata gcagttcttg caaggaagca 9180 aagtttttca acggtttgag accgtccgcc gtaggcatgc ttttgagcgt ttgaccaagc 9240 agt tccaggc ggtcccacag ctcggtcacc tgctctacgg catctcgatc cagcatatct 9300 cctcgtttcg cgggttgggg cggctttcgc tgtacggcag tagtcggtgc tcgtccagac 9360 gggccagggt catgtctttc cacgggcgca gggtcctcgt cagcgtagtc tgggtcacgg 9420 tgaaggggtg cgctccgggc tgcgcgctgg ccagggtgcg cttgaggctg gtcctgctgg 9480 tgctgaagcg ctgccggtct tcgccctgcg cgtcggccag gtagcatttg accatggtgt 9540 catagtccag cccctccgcg gcgtggccct tggcgcgcag cttgcccttg gaggaggcgc 9600 cgcacgaggg gcagtgcaga cttttgaggg cgtagagctt gggcgcgaga aataccgatt 9660 ccggggagta ggcatccgcg ccgcaggccc cgcagacggt ctcgcattcc acgagccagg 9720 tgagctctgg ccgttcgggg tcaaaaacca ggtttccccc atgctttttg atgcgtttct 9780 tacctctggt ttccatgagc cggtgtccac gctcggtgac gaaaaggctg tccgtgtccc 9840 cgtatacaga ct tgagaggc ctgtcctcga gcggtgttcc gcggtcctcc tcgtatagaa 9900 actcggacca ctctgagaca aaggctcgcg tccaggccag cacgaaggag gctaagtggg 9960 aggggtagcg gtcgttgtcc actagggggt ccactcgctc cagggtgtga agacacatgt 10020 cgccctcttc ggcatcaagg aaggtgattg gtttgtaggt gtaggccacg tgaccgggtg 10080 ttcctgaagg ggggctataa aagggggtgg gggcgcgttc gtcctcactc tcttccgcat 10140 cgctgtctgc gagggccagc tgttggggtg agtactccct ctgaaaagcg ggcatgactt 10200 -64 -

201207108 ctgcgctaag attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt gagggtggcc gcatccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttggtggc aaacgacccg tagagggcgt tggacagcaa cttggcgatg gagcgcaggg tttggttttt gtcgcgatcg gcgcgctcct tggccgcgat gtttagctgc acgtattcgc gcgcaacgca ccgccattcg ggaaagacgg tggtgcgctc gtcgggcacc aggtgcacgc gccaaccgcg gttgtgcagg gtgacaaggt caacgctggt ggctacctct ccgcgtaggc gctcgttggt ccagcagagg cggccgccct tgcgcgagca gaatggcggt agggggtcta gctgcgtctc gtccgggggg tctgcgtcca cggtaaagac cccgggcagc aggcgcgcgt cgaagtagtc tatcttgcat ccttgcaagt ctagcgcctg ctgccatgcg cgggcggcaa gcgcgcgctc gtatgggttg agtgggggac cccatggcat ggggtgggtg agcgcggagg cgtacatgcc gcaaatgtcg taaacgtaga ggggctctct gagtattcca agatatgtag ggtagcatct tccaccgcgg atgctggcgc gcacgtaatc gtatagttcg tgcgagggag cgaggaggtc gggaccgagg ttgctacggg cgggctgctc tgctcggaag actatctgcc tgaagatggc atgtgagttg gatgatatgg ttggacgctg gaagacgttg aagctggcgt ctgtgagacc taccgcgtca cgcacgaagg aggcgtagga gtcgcgcagc ttgttgacca gctcggcggt gacctgcacg tctagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg tccctttttt ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtactc ttggatcgga aacccgtcgg cctccgaacg gtaagagcct agcatgtaga actggttgac ggcctggtag gcgcagcatc ccttttctac gggtagcgcg tatgcctgcg cggccttccg gagcgaggtg tgggtgagcg caaaggtgtc cctgaccatg actttgaggt actggtattt gaagtcagtg tcgtcgcatc cgccctgctc ccagagcaaa aagtccgtgc gctttttgga acgcggattt ggcagggcga aggtgacatc gttgaagagt atctttcccg cgcgaggcat aaagttgcgt gtgatgcgga agggtcccgg cacctcggaa cggttgttaa ttacctgggc ggcgagcacg atctcgtcaa agccgttgat gttgtggccc acaatgtaaa gttccaagaa gcgcgggatg cccttgatgg aaggcaattt tttaagttcc tcgtaggtga gctcttcagg ggagctgagc ccgtgctctg aaagggccca gtctgcaaga tgagggttgg aagcgacgaa tgagctccac aggtcacggg ccattagcat ttgcaggtgg tcgcgaaagg tcctaaactg gcgacctatg gccatttttt ctggggtgat gcagtagaag gtaagcgggt cttgttccca gcggtcccat ccaaggttcg cggctaggtc tcgcgcggca gtcactagag gctcatctcc gccgaacttc atgaccagca tgaagggcac gagctgcttc ccaaaggccc ccatccaagt ataggtctct acatcgtagg tgacaaagag acgctcggtg cgaggatgcg agccgatcgg gaagaactgg atctcccgcc accaattgga ggagtggcta ttgatgtggt gaaagtagaa gtccctgcga cgggccgaac actcgtgctg gcttttgtaa aaacgtgcgc agtactggca gcggtgcacg ggctgtacat cctgcacgag gttgacctga cgaccgcgca caaggaagca gagtgggaat ttgagcccct cgcctggcgg gtttggctgg tggtcttcta cttcggctgc ttgtccttga ccgtctggct gctcgagggg agttacggtg gatcggacca ccacgccgcg cgagcccaaa gtccagatgt ccgcgcgcgg cggtcggagc ttgatgacaa 10260 10320 10380 10440 10500 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 11160 11220 11280 11340 11400 11460 11520 11580 11640 11700 11760 11820 11880 11940 12000 12060 12120 12180 12240 12300 32360 12420 -65 - 201207108 catcgcgcag atgggagctg tccatggtct ggagctcccg cggcgtcagg tcaggcggga 12480 gctcctgcag gtttacctcg catagacggg tcagggcgcg ggctagatcc aggtgatacc 12540 taatttccag gggctggttg gtggcggcgt cgatggcttg Caagaggccg catccccgcg 12600 gcgcgactac ggtaccgcgc ggcgggcggt gggccgcggg ggtgtccttg gatgatgcat 12660 ctaaaagcgg tgacgcg ggc gagcccccgg aggtaggggg ggctccggac ccgccgggag 12720 agggggcagg ggcacgtcgg cgccgcgcgc gggcaggagc tggtgctgcg cgcgtaggtt 12780 gctggcgaac gcgacgacgc ggcggttgat ctcctgaatc tggcgcctct gcgtgaagac 12840 gacgggcccg gtgagcttga acctgaaaga gagttcgaca gaatcaattt cggtgtcgtt 12900 gacggcggcc tggcgcaaaa tctcctgcac gtctcctgag ttgtcttgat aggcgatctc 12960 ggccatgaac tgctcgatct cttcctcctg gagatctccg cgtccggctc gctccacggt 13020 ggcggcgagg tcgttggaaa tgcgggccat gagctgcgag aaggcgttga ggcctccctc 13080 gttccagacg cggctgtaga ccacgccccc ttcggcatcg cgggcgcgca tgaccacctg 13140 cgcgagattg agctccacgt gccgggcgaa gacggcgtag tttcgcaggc gctgaaagag 13200 gtagttgagg gtggtggcgg tgtgttctgc cacgaagaag tacataaccc agcgtcgcaa 13260 cgtggattcg ttgatatccc ccaaggcctc aaggcgctcc atggcctcgt agaagtccac 13320 ggcgaagttg aaaaactggg agttgcgcgc cgacacggtt aactcctcct ccagaagacg 13380 gatgagctcg gcgacagtgt cgcgcacctc gcgctcaaag gctacagggg cctcttcttc 13440 ttcttcaatc tcctcttcca taagggcctc cccttcttct tcttctggcg gcggtggggg 135 00 aggggggaca cggcggcgac gacggcgcac cgggaggcgg tcgacaaagc gctcgatcat 13560 ctccccgcgg cgacggcgca tggtctcggt gacggcgcgg ccgttctcgc gggggcgcag 13620 ttggaagacg ccgcccgtca tgtcccggtt atgggttggc ggggggctgc catgcggcag 13680 ggatacggcg ctaacgatgc atctcaacaa ttgttgtgta ggtactccgc cgccgaggga 13740 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc 13800 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggcggcggt cggggttgtt 13860 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt 13920 cgacagaagc accatgtcct tgggtccggc ctgctgaatg cgcaggcggt cggccatgcc 13980 ccaggcttcg ttttgacatc ggcgcaggtc tttgtagtag tcttgcatga gcctttctac 14040 cggcacttct tcttctcctt cctcttgtcc tgcatctctt gcatctatcg ctgcggcggc 14100 ggcggagttt ggccgtaggt ggcgccctct tcctcccatg cgtgtgaccc cgaagcccct 14160 catcggctga agcagggcta ggtcggcgac aacgcgctcg gctaatatgg cctgctgcac 14220 ctgcgtgagg gtagactgga agtcatccat gtccacaaag cggtggtatg cgcccgtgtt 14280 gatggtgtaa gtgcagttgg ccataacgga ccagttaacg gtc tggtgac ccggctgcga 14340 gagctcggtg tacctgagac gcgagtaagc cctcgagtca aatacgtagt cgttgcaagt 14400 ccgcaccagg tactggtatc ccaccaaaaa gtgcggcggc ggctggcggt agaggggcca 14460 gcgtagggtg gccggggctc cg £ gggcgag atcttccaac ataaggcgat gatatccgta 14520 gatgtacctg gacatccagg tgatgccggc ggcggtggtg gaggcgcgcg gaaagtcgcg 14580 gacgcggttc cagatgttgc gcagcggcaa aaagtgctcc atggtcggga cgctctggcc 14640 ggtcaggcgc gcgcaatcgt tgacgctcta gcgtgcaaaa ggagagcctg taagcgggca 14700 -66 - 201207108

ctcttccgtg gccccgtatc tgtgcgacgt ctgcgctagc aagcattaag gacccccggt catgcaagac cccagatgca agcagcggca ccgcggttga acctggactt acccaagggt ttcgcgaccg gcgagctgcg acgcgcgaac catacgagca gtacgcttgt gcgcgctgga agcacagcag gccgctggct tgagcctggc acgcccgcaa ggttctacat gcaacgagcg agctgatgca agtcctactt cagctggggc tggaggaata tgtttctgat ccagccgtcc gctgactgcg aattctggaa cgtaaacgcg cgcgctgctt gctggtgggg cctgggctcc gggacaggag gtctggtgga cggccgtccg cagacaacgg ttttttggcc tggctcgctc tcgagtctcg cccgcttgca tccggtgctg gacatgcagg cgcggcagca ggaggagggc gcagctgaag cgagggagag gcatggcctg cgggattagt gacggtgaac ggcgcgcgag gcaaaaccca ggacaacgag gctcgatttg tgacaaggtg gatataccat gcgcatggcg catccacaag cagcctgcaa tgacgcgggc cggacctggg tgacgaggac cagatgatgc ggccttaact cgcaatcctg gcggtggtcc ctggccgaaa cagcgcgtgg gatgtgcgcg atggttgcac gactacacca taaattcgca ccgtgatcca gggagtgctc actggccgcg cctgtagccg gaccggccgg aattcctccg cggcagatgc gcaccctccc gatggtgatt gagggcctgg cgtgatacgc gagcccgagg aatcgcgagc cccgcgcgcg caggagatta gaggtggcta aatagcaagc gcattcaggg ataaacatcc gccgccatca accccttacg ctgaaggtgc gccgtgagcg agggccctgg gctgacctgc ctggcggtgg gatgagtacg aagacgcaac ccacggacga acgcgttccg cggcgcgcgc acagggccat ctcgttacaa aggccgtggc taaacgcctt actttgtgag agggtatcat tgcggttacc cttttggctt cgcagcgtaa gagggttatt actgcggcga gaaacaggga gcccccctcc ctcctcctac acgaaccccc cgcggctagg gtgaggcgta agatgcggga ggttgctgcg cacacgtggc actttcaaaa taggactgat cgctcatggc atgcgctgct tgcagagcat actattccat ttcccataga ttaccttgag tgagccggcg ctggcacggg gctgggcccc cacccgcgcg agccagagga ggacccggcg ctggcgccag gcagcagccg aaaccccacg ccggcccgac cagcggcaac gcagcgtgag cctgagtaca cgcactgcgg ggcggacgac gcccgcgtgt ccttccaggc gcggttaggc ttccaagggt acgggggttt cgagcccctt tcagcagcgg cgcgtcagga gcggcgccgg agcgccctct cgtgccgcgg tcgaaagttc cgaggaggac ggccgccgac aagctttaac gcatctgtgg gcagctgttc aaacatagta agtggtgcag gcttagcctg caaggaggta cgacgacctg gcgcgagctc cagcggcgat aagccgacgc cgctggcaac cggcgagtac gtgcgggcgg gtcatggacc caggccaacc cacgagaagg gaggccggcc gtgcagacc a cgcgcgcagc cagcccgcca ctaatggtga cggggttcga cgaacccagg gcggcggctg tggaaagcga tgagtcgcgg gcctccccgt ttttgctttt caagagcaag ggggcgacat gcccggcact cctgagcggc cagaacctgt cacgcagggc tttgagcccg ctggtaaccg aaccacgtgc gactttgtaa cttatagtgc gagcccgagg gagcgcagct ggcaagtttt aagatcgagg ggcgtttatc agcgaccgcg agagaggccg gccctggagg gtcggcggcg taagcggtga cgctgcagag gcatcatgtc ggctctccgc tgctggcgat tggtctacga acctggaccg agcagggcaa acgtgccgcg ctgagacacc 14760 14820 14880 14940 15000 15060 15120 15180 15240 15300 15360 15420 15480 15540 15600 15660 15720 15780 15840 15900 15960 16020 16080 16140 16200 16260 16320 16380 16440 16500 16560 16620 16680 16740 16800 16860 16920 -67 - 201207108 gcaaagtgag gtgtaccagt ctgggccaga ctattttttc cagaccagta gacaaggcct 16980 gcagaccgta aacctgagcc aggctttcaa aaacttgcag gggctgtggg gggtgcgggc 17040 tcccacaggc gaccgcgcga ccgtgtctag cttgctgacg Cccaactcgc gcctgttgct 17100 gctgctaata gcgcccttca cggacagtgg cagcgtgtcc cgggacacat acctaggtca 17160 cttgctgaca ctgtaccgcg aggcca tagg tcaggcgcat gtggacgagc atactttcca 17220 ggagattaca agtgtcagcc gcgcgctggg gcaggaggac acgggcagcc tggaggcaac 17280 cctaaactac ctgctgacca accggcggca gaagatcccc tcgttgcaca gtttaaacag 17340 cgaggaggag cgcattttgc gctacgtgca gcagagcgtg agccttaacc tgatgcgcga 17400 cggggtaacg cccagcgtgg cgctggacat gaccgcgcgc aacatggaac cgggcatgta 17460 tgcctcaaac cggccgttta tcaaccgcct aatggactac ttgcatcgcg cggccgccgt 17520 gaaccccgag tatttcacca atgccatctt gaacccgcac tggctaccgc cccctggttt 17580 ctacaccggg ggattcgagg tgcccgaggg taacgatgga ttcctctggg acgacataga 17640 cgacagcgtg ttttccccgc aaccgcagac cctgctagag ttgcaacagc gcgagcaggc 17700 agaggcggcg ctgcgaaagg aaagcttccg caggccaagc agcttgtccg atctaggcgc 17760 tgcggccccg cggtcagatg ctagtagccc atttccaagc ttgatagggt ctcttaccag 17820 cactcgcacc acccgcccgc gcctgctggg cgaggaggag tacctaaaca actcgctgct 17880 gcagccgcag cgcgaaaaaa acctgcctcc ggcatttccc aacaacggga tagagagcct 17940 agtggacaag atgagtagat ggaagacgta cgcgcaggag cacagggacg tgccaggccc 18000 gcgccc gccc acccgtcgtc aaaggcacga ccgtcagcgg ggtctggtgt gggaggacga 18060 tgactcggca gacgacagca gcgtcctgga tttgggaggg agtggcaacc cgtttgcgca 18120 cct tcgcccc aggctgggga gaatgtttta aaaaaaaaaa aagcatgatg caaaataaaa 18180 aactcaccaa ggccatggca ccgagcgttg gttttcttgt attcccctta gtatgcggcg 18240 cgcggcgatg tatgaggaag gtcctcctcc ctcctacgag agtgtggtga gcgcggcgcc 18300 agtggcggcg gcgctgggtt ctcccttcga tgctcccctg gacccgccgt ttgtgcctcc 18360 gcggtacctg cggcctaccg gggggagaaa cagcatccgt tactctgagt tggcacccct 18420 attcgacacc acccgtgtgt acctggtgga caacaagtca acggatgtgg catccctgaa 18480 ctaccagaac gaccacagca actttctgac cacggtcatt caaaacaatg actacagccc 18540 gggggaggca agcacacaga ccatcaatct tgacgaccgg tcgcactggg gcggcgacct 18600 gaaaaccatc ctgcatacca acatgccaaa tgtgaacgag ttcatgttta ccaataagtt 18660 taaggcgcgg gtgatggtgt cgcgcttgcc tactaaggac aatcaggtgg agctgaaata 18720 cgagtgggtg gagttcacgc tgcccgaggg caactactcc gagaccatga ccatagacct 18780 tatgaacaac gcgatcgtgg agcactactt gaaagtgggc agacagaacg g ggttctgga 18840 aagcgacatc ggggtaaagt ttgacacccg caacttcaga ctggggtttg accccgtcac 18900 tggtcttgtc atgcctgggg tatatacaaa cgaagccttc catccagaca tcattttgct 18960 gccaggatgc ggggtggact tcacccacag ccgcctgagc aacttgttgg gcatccgcaa 19020 gcggcaaccc ttccaggagg gctttaggat cacctacgat gatctggagg gtggtaacat 19080 tcccgcactg ttggatgtgg acgcctacca ggcgagcttg aaagatgaca ccgaacaggg 19140 Cgggggtggc gcaggcggca gcaacagcag tggcagcggc gcggaagaga • 68 - actccaacgc 19200 201207108

ggcagccgcg gcaatgcagc cggtggagga catgaacgat catgccattc gcggcgacac 19260 ctttgccaca cgggctgagg agaagcgcgc tgaggccgaa gcagcggccg aagctgccgc 19320 ccccgctgcg caacccgagg tcgagaagcc tcagaagaaa ccggtgatca aacccctgac 19380 agaggacagc aagaaacgca gttacaacct aataagcaat gacagcacct tcacccagta 19440 ccgcagctgg taccttgcat acaactacgg cgaccctcag accggaatcc gctcatggac 19500 cctgctttgc actcctgacg taacctgcgg ctcggagcag gtctactggt cgttgccaga 19560 catgatgcaa gaccccgtga ccttccgctc cacgcgccag atcagcaact ttccggtggt 19620 gggcgccgag ctgttgcccg tgcactccaa gagcttctac aacgaccagg ccgtctactc 19680 ccaactcatc cgccagttta cctctctgac ccacgtgitc aatcgctttc ccgagaacca 19740 gattttggcg cgcccgccag cccccaccat caccaccgtc agtgaaaacg ttcctgctct 19800 cacagatcac gggacgctac cgctgcgcaa cagcatcgga ggagtccagc gagtgaccat 19860 tactgacgcc agacgccgca cctgccccta cgtttacaag gccctgggca tagtctcgcc 19920 gcgcgtccta tcgagccgca ctttttgagc aagcatgtcc atccttatat cgcccagcaa 19980 taacacaggc tggggcctgc gcttcccaag caagatgttt Sgcgggg cca agaagcgctc 20040 cgaccaacac ccagtgcgcg tgcgcgggca ctaccgcgcg ccctggggcg cgcacaaacg 20100 cggccgcact gggcgcacca ccgtcgatga cgccatcgac gcggtggtgg aggaggcgcg 20160 caactacacg cccacgccgc caccagtgtc cacagtggac gcggccattc agaccgtggt 20220 gcgcggagcc cggcgctatg ctaaaatgaa gagacggcgg aggcgcgtag cacgtcgcca 20280 ccgccgccga cccggcactg ccgcccaacg cgcggcggcg gccctgctta accgcgcacg 20340 tcgcaccggc cgacgggcgg ccatgcgggc cgctcgaagg ctggccgcgg gtattgtcac 20400 tgtgcccccc aggtccaggc gacgagcggc cgccgcagca gccgcggcca ttagtgctat 20460 gactcagggt cgcaggggca acgtgtattg ggtgcgcgac tcggttagcg gcctgcgcgt 20520 gcccgtgcgc acccgccccc cgcgcaacta gattgcaaga aaaaactact tagactcgta 20580 ctgttgtatg tatccagcgg cggcggcgcg caacgaagct atgtccaagc gcaaaatcaa 20640 agaagagatg ctccaggtca tcgcgccgga gatctatggc cccccgaaga aggaagagca 20700 ggattacaag ccccgaaagc taaagcgggt caaaaagaaa aagaaagatg atgatgatga 20760 acttgacgac gaggtggaac tgctgcacgc taccgcgccc aggcgacggg tacagtggaa 20820 aggtcgacgc gtaaaacgtg ttttgcg acc cggcaccacc gtagtcttta cgcccggtga 20880 gcgctccacc cgcacctaca agcgcgtgta tgatgaggtg tacggcgacg aggacctgct 20940 tgagcaggcc aacgagcgcc tcggggagtt tgcctacgga aagcggcata aggacatgct 21000 ggcgttgccg ctggacgagg gcaacccaac acctagccta aagcccgtaa cactgcagca 21060 ggtgctgccc gcgcttgcac cgtccgaaga aaagcgcggc ctaaagcgcg agtctggtga 21120 cttggcaccc accgtgcagc tgatggtacc caagcgccag cgactggaag atgtcttgga 21180 aaaaatgacc gtggaacctg ggctggagcc cgaggtccgc gtgcggccaa tcaagcaggt 21240 ggcgccggga ctgggcgtgc agaccgtgga cgttcagata cccactacca gtagcaccag 21300 tattgccacc gccacagagg gcatggagac acaaacgtcc ccggttgcct cagcggtggc 21360 ggatgccgcg gtgcaggcgg tcgctgcggc cgcgtccaag acctctacgg aggtgcaaac 21420 -69 - 201207108 ggacccgtgg atgtttcgcg tttcagcccc ccggcgcccg cgccgttcga ggaagtacgg 21480 cgccgccagc gcgctactgc ccgaatatgc cctacatcct tccattgcgc ctacccccgg 21540 ctatcgtggc tacacctacc gccccagaag acgagcaact acccgacgcc gaaccaccac 21600 tggaacccgc cgccgccgtc gccgtcgcca gcccgtgctg gccccgattt ccgtgcgcag 21660 ggtggctcgc gaaggaggca ggaccctggt gctgccaaca gcgcgctacc accccagcat 21720 cgtttaaaag ccggtctttg tggttcttgc agatatggcc ctcacctgcc gcctccgttt 21780 cccggtgccg ggattccgag gaagaatgca ccgtaggagg ggcatggccg gccacggcct 21840 gacgggcggc atgcgtcgtg cgcaccaccg gcggcggcgc gcgtcgcacc gtcgcatgcg 21900 cggcggtatc ctgcccctcc ttattccact gatcgccgcg gcgattggcg ccgtgcccgg 21960 aattgcatcc gtggccttgc aggcgcagag acactgatta aaaacaagtt gcatgtggaa 22020 aaatcaaaat aaaaagtctg gactctcacg ctcgcttggt cctgtaacta ttttgtagaa 22080 tggaagacat caactttgcg tctctggccc cgcgacacgg ctcgcgcccg ttcatgggaa 22140 22440 agacagtgtc tccagagggg actggcaaga tatcggcacc agcaatatga gcggtggcgc cttcagctgg ggctcgctgt 22200 ggagcggcat taaaaatttc ggttccaccg ttaagaacta tggcagcaag gcctggaaca 22260 gcagcacagg ccagatgctg agggataagt tgaaagagca aaatttccaa caaaaggtgg 22320 tagatggcct ggcctctggc attagcgggg tggtggacct ggccaaccag gcagtgcaaa 22380 ataagattaa cagtaagctt gatccccgcc ctcccgtaga ggagcctcca ccggccgtgg cgtggcgaaa agcgtccgcg ccccgacagg gaagaaactc 22500 tggtgacgca aatagacgag cctccctcgt acgaggaggc actaaagcaa ggcctgccca 22560 ccacccgtcc catcgcgccc atggctaccg gagtgctggg ccagcacaca cccgtaacgc 22620 tggacctgcc tccccccgcc gacacccagc agaaacctgt gctgccaggc ccgaccgccg 22680 ttgttgtaac ccgtcctagc cgcgcgtccc tgcgccgcgc cgccagcggt ccgcgatcgt 22740 tgcggcccgt agccagtggc aactggcaaa gcacactgaa cagcatcgtg ggtctggggg 22800 tgcaatccct gaagcgccga cgatgcttct gatagctaac gtgtcgtatg tgtgtcatgt 22860 atgcgtccat gtcgccgcca gaggagctgc tgagccgccg cgcgcccgct ttccaagatg 22920 gctacccctt cgatgatgcc gcagtggtct tacatgcaca tctcgggcca ggacgcctcg 22980 gagtacctga gccccgggct ggtgcagttt gcccgcgcca ccgagacgta cttcagcctg 23040 aataacaagt ttagaaaccc cacggtggcg cctacgcacg acgtgaccac agaccggtcc 23100 cagcgtttga cgctgcggtt catccctgtg gaccgtgagg atactgcgta ctcgtacaag 23160 gcgcggttca ccctagctgt gggtgataac cgtgtgctgg acatggcttc cacgtacttt 23220 gacatccgcg gcgtgctgga caggggccct acttttaagc cctactctgg cactgcctac 23280 aacgccctgg ctcccaaggg tgccccaaat ccttgcgaat gggatgaagc tgctactgct 23340 cttgaaataa acctagaaga agaggacgat gacaacgaag acgaagtaga cgagcaagct 23400 gagcagcaaa aaactcacgt atttgggcag gcgccttatt ctggtataaa tattacaaag 23460 gagggtattc aaataggtgt cgaaggtcaa acacctaaat atgccgataa aacatttcaa 23520 cctgaacctc aaataggaga atctcagtgg tacgaaacag aaattaatca tgcagctggg 23580 agagtcctaa aaaagactac cccaatgaaa ccatgttacg gttcatatgc aaaacccaca 23640 aatgaaaatg gagggcaagg cattcttgta aagcaacaaa atggaaagct agaaagtcaa 23700 • 70 Ο

201207108 gtggaaatgc aatttttctc aactactgag gcagccgcag gcaatggtga taacttgact cctaaagtgg tattgtacag tgaagatgta gatatagaaa ccccagacac tcatatttct tacatgccca ctattaagga aggtaactca cgagaactaa tgggccaaca atctatgccc aacaggccta attacattgc ttttagggac aattttattg gtctaatgta ttacaacagc acgggtaata tgggtgttct ggcgggccaa gcatcgcagt tgaatgctgt tgtagatttg caagacagaa acacagagct ttcataccag cttttgcttg attccattgg tgatagaacc aggtactttt ctatgtggaa tcaggctgtt gacagctatg atccagatgt tagaattatt gaaaatcatg gaactgaaga tgaacttcca aattactgct ttccactggg aggtgtgatt aatacagaga ctcttaccaa ggtaaaacct aaaacaggtc aggaaaatgg atgggaaaaa gatgctacag aattttcaga taaaaatgaa ataagagttg gaaataattt tgccatggaa atcaatctaa atgccaacct gtggagaaat ttcctgtact ccaacatagc gctgtatttg cccgacaagc taaagtacag tccttccaac gtaaaaattt ctgataaccc aaacacctac gactacatga acaagcgagt ggtggctccc gggctagtgg actgctacat taaccttgga gcacgctggt cccttgacta tatggacaac gtcaacccat ttaaccacca ccgcaatgct ggcctgcgct accgctcaat gttgctgggc aatggtcgct atgtgccctt ccacatccag gtgcctcaga agttctttgc cattaaaaac ctccttctcc tgccgggctc atacacctac gagtggaact tcaggaagga tgttaacatg gttctgcaga gctccctagg aaatgaccta agggttgacg gagccagcat taagtttgat agcatttgcc tttacgccac cttcttcccc atggcccaca acaccgcctc cacgcttgag gccatgctta gaaacgacac caacgaccag tcctttaacg actatctctc cgccgccaac atgctctacc ctatacccgc caacgctacc aacgtgccca tatccatccc ctcccgcaac tgggcggctt tccgcggctg ggccttcacg cgccttaaga ctaaggaaac cccatcactg ggctcgggct acgaccctta ttacacctac tctggctcta taccctacct agatggaacc ttttacctca accacacctt taagaaggtg gccattacct ttgactcttc tgtcagctgg cctggcaatg accgcctgct tacccccaac gagtttgaaa ttaagcgctc agttgacggg gagggttaca acgttgccca gtgtaacatg accaaagact ggttcctggt acaaatgcta gciaactata acattggcta ccagggcttc tatatcccag agagctacaa ggaccgcatg tactccttct ttagaaactt ccagcccatg agccgtcagg tggtggatga tactaaatac aaggactacc aacaggtggg catcctacac caacacaaca actctggatt tgttggctac cttgccccca ccatgcgcga aggacaggcc taccctgcta acttccccta tccgcttata ggcaagaccg cagttgacag cattacccag aaaaagtttc tttgcgatcg caccctttgg cgcatcccat tctccagtaa ctttatgtcc atgggcgcac tcacagacct gggccaaaac cttctctacg ccaactccgc ccacgcgcta gacatgactt ttgaggtgga tcccatggac gagcccaccc ttctttatgt tttgtttgaa gtctttgacg tggtccgtgt gcaccagccg caccgcggcg tcatcgaaac cgtgtacctg cgcacgccct tctcggccgg caacgccaca acataaagaa gcaagcaaca tcaacaacag ctgccgccat gggctccagt gagcaggaac tgaaagccat tgtcaaagat cttggttgtg ggccatattt tttgggcacc tatgacaagc gctttccagg ctttgtttct ccacacaagc 23760 23820 23880 23940 24000 24060 24120 24180 24240 24300 24360 24420 24480 24540 24600 24660 24720 24780 24840 24900 24960 25020 25080 25140 25200 25260 25320 25380 25440 25500 25560 25620 25680 25740 25800 25860 25920 201207108 tcgcctgcgc catagtcaat acggccggtc gcgagactgg gggcgtacac tggatggcct 25980 ttgcctggaa cccgcacica aaaacatgct acctctttga gccctttggc ttttctgacc 26040 agcgactcaa gcaggtttac cagtttgagt acgagtcact cctgcgccgt agcgccattg 26100 Cttcttcccc cgaccgctgt ataacgctgg aaaagtccac ccaaagcgta caggggccca 26160 actcggccgc ctgtggacta tt ctgctgca tgtttctcca cgcctttgcc aactggcccc 26220 aaactcccat ggatcacaac cccaccatga acct tat tac cggggtaccc aactccatgc 26280 tcaacagtcc ccaggtacag cccaccctgc gtcgcaacca ggaacagctc tacagcttcc 26340 tggagcgcca ctcgccctac ttccgcagcc acagtgcgca gattaggagc gccacttctt 26400 tttgtcactt gaaaaacatg taaaaataat gtactagaga cactttcaat aaaggcaaat 26460 gcttttattt gtacactcic gggtgaiiat t tacccccac cct tgccgtc tgcgccgttt 26520 aaaaatcaaa ggggttctgc cgcgcatcgc tatgcgccac tggcagggac acgttgcgat 26580 actggtgttt agtgctccac ttaaactcag gcacaaccat ccgcggcagc tcggtgaagt 26640 tttcactcca caggctgcgc accatcacca acgcgtttag caggtcgggc gccgatatct 26700 tgaagtcgca gttggggcct ccgccctgcg cgcgcgagtt gcgatacaca gggttgcagc 26760 actggaacac tatcagcgcc gggtggtgca cgctggccag cacgctcttg tcggagatca 26820 gatccgcgtc caggtcctcc gcgttgctca gggcgaacgg agtcaacttt ggtagctgcc 26880 ttcccaaaaa gggcgcgtgc ccaggctttg agttgcactc gcaccgtagt ggcatcaaaa 26940 ggtgaccgtg cccggtctgg gcgttaggat acagcgcctg cataaaagcc ttgatctgct 27000 taaaagccac ctgagccttt gcgccttcag agaagaacat gccgcaagac ttgccggaaa 27060 actgattggc cggacaggcc gcgtcgtgca cgcagcacct tgcgtcggtg 27120 gcaccacatt tcggccccac cggttcttca cgatcttggc cttgctagac tgctccttca 27180 gcgcgcgctg cccgttttcg ctcgtcacat ccatttcaat cacgtgctcc 27240 taatgcttcc gtgtagacac ttaagctcgc cttcgatctc agcgcagcgg tgcagccaca 27300 acgcgcagcc cgtgggctcg tgatgcttgt aggtcacctc tgcaaacgac tgcaggtacg 27360 cctgcaggaa tcgccccatc atcgtcacaa aggtcttgtt gctggtgaag gtcagctgca 27420 acccgcggtg ttggagatct ttatttatca ctcctcgttc agccaggtct tgcatacggc cgccagagct tccacttggt 27480 caggcagtag tttgaagttc gcctttagat cgttatccac gtggtacttg tccatcagcg 27540 cgcgcgcagc ctccatgccc ttctcccacg cagacacgat cggcacactc agcgggttca 27600 tcaccgtaat ttcactttcc gcttcgctgg gctcttcctc ttcctcttgc gtccgcatac 27660 cacgcgccac tgggtcgtct tcattcagcc gccgcactgt gcgcttacct cctttgccat 27720 gcttgattag caccggtggg ttgctgaaac ccaccatttg tagcgccaca tcttctcttt 27780 cttcctcgct gtccacgatt acctctggtg atggcgggcg ctcgg gcttg ggagaagggc 27840 gcttcttttt cttcttgggc gcaatggcca aatccgccgc cgaggtcgat ggccgcgggc 27900 tgggtgtgcg cggcaccagc gcgtcttgtg atgagtcttc ctcgtcctcg gactcgatac 27960 gccgcctcat ccgctttttt gggggcgccc ggggaggcgg cggcgacggg gacggggacg 28020 acacgtcctc catggttggg ggacgtcgcg ccgcaccgcg tccgcgctcg ggggtggttt 28080 cgcgctgctc ctcttcccga ctggccattt ccttctccta taggcagaaa aagatcatgg 28140 agtcagtcga gaagaaggac agcctaaccg ccccctctga gttcgccacc -72 - accgcctcca 28200 201207108

ccgatgccgc caacgcgcct accaccttcc ccgtcgaggc acccccgctt gaggaggagg 28260 aagtgattat cgagcaggac ccaggttttg taagcgaaga cgacgaggac cgctcagtac 28320 caacagagga taaaaagcaa gaccaggaca acgcagaggc aaacgaggaa caagtcgggc 28380 ggggggacga aaggcatggc gactacctag atgtgggaga cgacgtgctg ttgaagcatc 28440 tgcagcgcca gtgcgccatt atctgcgacg cgttgcaaga gcgcagcgat gtgcccctcg 28500 ccatagcgga tgtcagcctt gcctacgaac gccacctatt ctcaccgcgc gtacccccca 28560 aacgccaaga aaacggcaca tgcgagccca acccgcgcct caacttctac cccgtatttg 28620 ccgtgccaga ggtgcttgcc acctatcaca tctttttcca aaactgcaag atacccctat 28680 cctgccgtgc caaccgcagc cgagcggaca agcagctggc cttgcggcag ggcgctgtca 28740 tacctgatat cgcctcgctc aacgaagtgc caaaaatctt tgagggtctt ggacgcgacg 28800 agaagcgcgc ggcaaacgct ctgcaacagg aaaacagcga aaatgaaagt cactctggag 28860 tgttggtgga actcgagggt gacaacgcgc gcctagccgt actaaaacgc agcatcgagg 28920 tcacccactt tgcctacccg gcact taacc taccccccaa ggtcatgagc acagtcatga 28980 gtgagctgat cgtgcgccgt gcgcagcccc tggagaggga tgcaaa tttg caagaacaaa 29040 cagaggaggg cctacccgca gttggcgacg agcagctagc gcgctggctt caaacgcgcg 29100 agcctgccga cttggaggag cgacgcaaac taatgatggc cgcagtgctc gttaccgtgg 29160 agcttgagtg catgcagcgg ttctttgctg acccggagat gcagcgcaag ctagaggaaa 29220 cat tgcacta cacctttcga cagggctacg tacgccaggc ctgcaagatc tccaacgtgg 29280 agctctgcaa cctggtctcc taccttggaa tt ttgcacga aaaccgcctt gggcaaaacg 29340 tgcttcattc cacgctcaag ggcgaggcgc gccgcgacta cgtccgcgac tgcgtttact 29400 tatttctatg ctacacctgg cagacggcca tgggcgtttg gcagcagtgc ttggaggagt 29460 gcaacctcaa ggagctgcag aaactgctaa agcaaaactt gaaggaccta tggacggcct 29520 tcaacgagcg ctccgtggcc gcgcacctgg cggacatcat tttccccgaa cgcctgctta 29580 aaaccctgca acagggtctg ccagacttca ccagtcaaag catgttgcag aactttagga 29640 actttatcct agagcgctca ggaatcttgc ccgccacctg ctgtgcactt cctagcgact 29700 ttgtgcccat taagtaccgc gaatgccctc cgccgctttg gggccactgc taccttctgc 29760 agctagccaa ctaccttgcc taccactctg acataatgga agacgtgagc ggtgacggtc 29820 tactggagtg tcactgtcgc tgca acctat gcaccccgca ccgctccctg gtttgcaatt 29880 cgcagctgct taacgaaagt caaattatcg gtacctttga gctgcagggt ccctcgcctg 29940 acgaaaagtc cgcggctccg gggttgaaac tcactccggg gctgtggacg tcggcttacc 30000 ttcgcaaatt tgtacctgag gactaccacg cccacgagat taggttctac gaagaccaat 30060 cccgcccgcc taatgcggag cttaccgcct gcgtcattac ccagggccac attcttggcc 30120 aattgcaagc catcaacaaa gcccgccaag agtttctgct acgaaaggga cggggggttt 30180 acttggaccc ccagtccggc gaggagctca acccaatccc cccgccgccg cagccctatc 30240 agcagcagcc gcgggccctt gcttcccagg atggcaccca aaaagaagct gcagctgccg 30300 ccgccaccca cggacgagga ggaatactgg gacagtcagg cagaggaggt tttggacgag 30360 gaggaggagg acatgatgga agactgggag agcctagacg aggaagcttc cgaggtcgaa 30420 -73 - 201207108 gaggtgtcag acgaaacacc gtcaccctcg gtcgcattcc cctcgccggc gccccagaaa 30480 tcggcaaccg gttccagcat ggctacaacc tccgctcctc aggcgccgcc ggcactgccc 30540 gttcgccgac ccaaccgtag atgggacacc actggaacca gggccggtaa gtccaagcag 30600 ccgccgccgt tagcccaaga gcaacaacag cgccaaggct accgctcatg gcgcggg cac 30660 aagaacgcca tagttgcttg cttgcaagac tgtgggggca acatctcctt cgcccgccgc 30720 tttcttctct accatcacgg cgtggccttc ccccgtaaca tcctgcatta ctaccgtcat 30780 ctctacagcc catactgcac cggcggcagc ggcagcaaca gcagcggcca cacagaagca 30840 aaggcgaccg gatagcaaga ctctgacaaa gcccaagaaa tccacagcgg cggcagcagc 30900 aggaggagga gcgctgcgtc tggcgcccaa cgaacccgta tcgacccgcg agcttagaaa 30960 caggattttt cccactctgt atgctatatt tcaacagagc aggggccaag aacaagagct 31020 gaaaataaaa aacaggtctc tgcgatccct cacccgcagc tgcctgtatc acaaaagcga 31080 agatcagctt cggcgcacgc tggaagacgc ggaggctctc ttcagtaaat actgcgcgct 31140 gactcttaag gactagtttc gcgccctttc tcaaatttaa gcgcgaaaac tacgtcatct 31200 ccagcggcca cacccggcgc cagcacctgt tgtcagcgcc attatgagca aggaaattcc 31260 cacgccctac atgtggagtt accagccaca aatgggactt gcggctggag ctgcccaaga 31320 ctactcaacc cgaataaact acatgagcgc gggaccccac atgatatccc gggtcaacgg 31380 aatacgcgcc caccgaaacc gaat tctcct ggaacaggcg gctattacca ccacacctcg 31440 taataacctt aatccccgta gttggcccgc tgccct ggtg taccaggaaa gtcccgctcc 31500 caccactgtg gtacttccca gagacgccca ggccgaagtt cagatgacta actcaggggc 31560 gcagcttgcg ggcggctttc gtcacagggt gcggtcgccc gggcagggta taactcacct 31620 gacaatcaga gggcgaggta ttcagctcaa cgacgagtcg gtgagctcct cgcttggtct 31680 ccgtccggac gggacatttc agatcggcgg cgccggccgc tcttcattca cgcctcgtca 31740 ggcaatccta actctgcaga cctcgtcctc tgagccgcgc tctggaggca ttggaactct 31800 gcaatttatt gaggagtttg tgccatcggt ctactttaac cccttctcgg gacctcccgg 31860 ccactatccg gatcaattta ttcctaactt tgacgcggta aaggactcgg cggacggcta 31920 cgactgaatg ttaagtggag aggcagagca actgcgcctg aaacacctgg tccactgtcg 31980 ccgccacaag tgctttgccc gcgactccgg tgagttttgc tactttgaat tgcccgagga 32040 tcatatcgag ggcccggcgc acggcgtccg gcttaccgcc cagggagagc ttgcccgtag 32100 cctgattcgg gagtttaccc agcgccccct gctagttgag cgggacaggg gaccctgtgt 32160 tctcactgtg atttgcaact gtcctaaccc tggattacat caagatctta ttccctttaa 32220 ctaataaaaa aaaataataa agcatcactt acttaaaatc agttagcaaa tttctgtcca 32280 gtttattcag cagcac ctcc ttgccctcct cccagctctg gtattgcagc ttcctcctgg 32340 ctgcaaactt tctccacaat ctaaatggaa tgtcagtttc ctcctgttcc tgtccatccg 32400 cacccactat cttcatgttg ttgcagatga agcgcgcaag accgtctgaa gataccttca 32460 accccgtgta tccatatgac acggaaaccg gtcctccaac tgtgcctttt cttactcctc 32520 cctttgtatc ccccaatggg tttcaagaga gtccccctgg ggtactctct ttgcgcctat 32580 ccgaacctct agttacctcc aatggcatgc ttgcgctcaa aatgggcaac ggcctctctc 32640 tggacgaggc cggcaacctt acctcccaaa atgtaaccac tgtgagccca cctctcaaaa 32700 -74 -

C

201207108 aaaccaagtc aaacataaac ctggaaatat ctgcacccct cacagttacc tcagaagccc taactgtggc tgccgccgca ccictaatgg tcgcgggcaa cacactcacc atgcaatcac aggccccgct aaccgtgcac gactccaaac ttagcattgc cacccaagga cccctcacag tgtcagaagg aaagctagcc ctgcaaacat caggccccct caccaccacc gatagcagta cccttactat cactgcctca ccccctctaa ctactgccac tggtagcttg ggcattgact tgaaagagcc catttataca caaaatggaa aactaggact aaagtacggg gctcctttgc atgtaacaga cgacctaaac actttgaccg tagcaactgg tccaggtgtg actattaata atacttcctt gcaaactaaa gttactggag ccttgggttt tgattcacaa ggcaatatgc aacttaatgt agcaggagga ctaaggattg attctcaaaa cagacgcctt atacttgatg ttagttatcc gtttgatgct caaaaccaac taaatctaag actaggacag ggccctctit ttataaactc agcccacaac ttggatatta actacaacaa aggcctttac ttgtttacag cttcaaacaa ttccaaaaag cttgaggtta acctaagcac tgccaagggg ttgatgtttg acgctacagc catagccatt aatgcaggag atgggcttga atttggttca cctaatgcac caaacacaaa tcccctcaaa acaaaaattg gccatggcct agaatttgat tcaaacaagg ctatggttcc taaactagga actggcctta gttttgacag cacaggtgcc attacagtag gaaacaaaaa taatgataag ctaactttgt ggaccacacc agctccatct cctaactgta gactaaatgc agagaaagat gctaaactca ctttggtctt aacaaaatgt ggcagtcaaa tacttgctac agtttcagtt ttggctgtta aaggcagttt ggctccaata tctggaacag ttcaaagtgc tcatcttatt ataagatttg acgaaaatgg agtgctacta aacaattcct tcctggaccc agaatattgg aactttagaa atggagatct tactgaaggc acagcctata caaacgctgt tggatttatg cctaacctat cagcttatcc aaaatctcac ggtaaaactg ccaaaagtaa cattgtcagt caagtttact taaacggaga caaaactaaa cctgtaacac taaccattac actaaacggt acacaggaaa caggagacac aactccaagt gcatactcta tgtcattttc atgggactgg tctggccaca actacattaa tgaaatattt gccacatcct cttacacttt ttcatacatt gcccaagaat aaagaatcgt ttgtgttatg tttcaacgtg tttatttttc aattgcagaa aatttcaagt catttttcat tcagtagtat agccccacca ccacatagct tatacagatc accgtacctt aatcaaactc acagaaccct agtattcaac ctgccacctc cctcccaaca cacagagtac acagtccttt ctccccggct ggccttaaaa agcatcatat catgggtaac agacatattc ttaggtgtta tattccacac ggtttcctgt cgagccaaac gctcatcagt gatattaata aactccccgg gcagctcact taagttcatg tcgctgtcca gctgctgagc cacaggctgc tgtccaactt gcggttgctt aacgggcggc gaaggagaag tccacgccta catgggggta gagtcataat cgtgcatcag gatagggcgg tggtgctgca gcagcgcgcg aataaactgc tgccgccgcc gctccgtcct gcaggaatac aacatggcag tggtctcctc agcgatgatt cgcaccgccc gcagcataag gcgccttgtc ctccgggcac agcagcgcac cctgatctca cttaaatcag cacagtaact gcagcacagc accacaatat tgttcaaaat cccacagtgc aaggcgctgt atccaaagct catggcgggg accacagaac ccacgtggcc atcataccac aagcgcaggt agattaagtg gcgacccctc 32760 32820 32880 32940 33000 33060 33120 33180 33240 33300 33360 33420 33480 33540 33600 33660 33720 33780 33840 33900 33960 34020 34080 34140 34200 34260 34320 34380 34440 34500 34560 34620 34680 34740 34800 34860 34920 -75 - 201207108 ataaacacgc tggacataaa cattacctct tttggcatgt tgtaattcac cacctcccgg 34980 taccatataa acctctgatt aaacatggcg ccatccacca ccatcctaaa ccagctggcc 35040 aaaacctgcc cgccggctat acactgcagg gaaccgggac Tggaacaatg acagtggaga 35100 gcccaggact cgtaaccatg gatcatcatg ctcgtcatga tatcaatgtt ggcacaacac 35160 aggcacacgt gcataca ctt cctcaggatt acaagctcct cccgcgttag aaccatatcc 35220 cagggaacaa cccattcctg aatcagcgta aatcccacac tgcagggaag acctcgcacg 35280 taactcacgt tgtgcattgt caaagtgtta cattcgggca gcagcggatg atcctccagt 35340 atggtagcgc gggtttctgt ctcaaaagga ggtagacgat ccctactgta cggagtgcgc 35400 cgagacaacc gagatcgtgt tggtcgtagt gtcatgccaa atggaacgcc ggacgtagtc 35460 atatttcctg aagcaaaacc aggtgcgggc gtgacaaaca gatctgcgtc tccggtctcg 35520 ccgcttagat cgctctgtgt agtagttgta gtatatccac tctctcaaag catccaggcg 35580 ccccctggct tcgggttcta tgtaaactcc ttcatgcgcc gctgccctga taacatccac 35640 caccgcagaa taagccacac ccagccaacc tacacattcg ttctgcgagt cacacacggg 35700 aggagcggga agagctggaa gaaccatgtt ttttttttta ttccaaaaga ttatccaaaa 35760 cctcaaaatg aagatctatt aagtgaacgc gctcccctcc ggtggcgtgg tcaaactcta 35820 cagccaaaga acagataatg gcatttgtaa gatgttgcac aatggcttcc aaaaggcaaa 35880 cggccctcac gtccaagtgg acgtaaaggc taaacccttc agggtgaatc tcctctataa 35940 acattccagc accttcaacc atgcccaaat aattctcatc tcgccacctt ctcaatatat 360 00 ctctaagcaa atcccgaata ttaagtccgg ccattgtaaa aatctgctcc agagcgccct 36060 ccaccttcag cctcaagcag cgaatcatga ttgcaaaaat tcaggttcct cacagacctg 36120 tataagattc aaaagcggaa cattaacaaa aataccgcga tcccgtaggt cccttcgcag 36180 ggccagctga acataatcgt gcaggtctgc acggaccagc gcggccactt ccccgccagg 36240 aaccatgaca aaagaaccca cactgat tat gacacgcata ctcggagcta tgctaaccag 36300 cgtagccccg atgtaagctt gttgcatggg cggcgatata aaatgcaagg tgctgctcaa 36360 aaaatcaggc aaagcctcgc gcaaaaaaga aagcacatcg tagtcatgct catgcagata 36420 aaggcaggta agctccggaa ccaccacaga aaaagacacc atttttctct caaacatgtc 36480 tgcgggtttc tgcataaaca caaaataaaa taacaaaaaa acatttaaac attagaagcc 36540 tgtcttacaa caggaaaaac aacccttata agcataagac ggactacggc catgccggcg 36600 tgaccgtaaa aaaactggtc accgtgatta aaaagcacca ccgacagctc ctcggtcatg 36660 tccggagtca taatgtaaga ctcggtaaac acatcaggtt gattcacatc ggtcagtgct 36720 aaaaagcgac cgaaatagcc cgggggaata catacccgca ggcgtagaga caacattaca 36780 gcccccatag gaggtataac aaaattaata ggagagaaaa ac acataaac acctgaaaaa 36840 ccctcctgcc taggcaaaat agcaccctcc cgctccagaa caacatacag cgcttccaca 36900 gcggcagcca taacagtcag ccttaccagt aaaaaagaaa acctattaaa aaaacaccac 36960 tcgacacggc accagctcaa tcagtcacag tgtaaaaaag ggccaagtgc agagcgagta 37020 tatataggac taaaaaatga cgtaacggtt aaagtccaca aaaaacaccc agaaaaccgc 37080 acgcgaacct acgcccagaa acgaaagcca aaaaacccac aacttcctca aatcgtcact 37140 tccgttttcc cacgttacgt cacttcccat tt taagaaaa ctacaattcc caacacatac 37200 -76 - 201207108 Aagttactcc gccctaaaac ctacgtcacc cgccccgttc ccacgccccg cgccacgtca caaactccac cccctcatta tcatattggc ttcaatccaa aataaggtat attattgatg atg 37260 37320 37323 &lt;210> 30 &lt;211&gt; 169 &lt;212&gt; DNA &lt;213>Artificial sequence &lt;220&gt;&lt;223> Human tumor necrosis factor 5· UTR &lt;400&gt; 30 ctccctcagc aaggacagca gaggaccagc taagagggag agaagcaact acagaccccc cctgaaaaca accctcagac gccacatccc ctgacaagct gccaggcagg ttctcttcct ctcacatact gacccacggc tccaccctct ctcccctgga aaggacacc 60 120 169

&lt;210&gt; 31 &lt;211&gt; 179 &lt;212&gt;212&gt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; 5u2 &lt;400&gt; 31 cgaagaaggt gagtaatctt aacatgctct tttttttttt ttttgctaat cccttttgtg tgctgatgtt aggatgacat ttacaacaaa tgtttgttcc tgacaggaaa aaccttgctg ggtaccttcg ttgccggaca cttcttgtcc Tctactttgg aaaaaaggaa ttgagagcc 60 120 179 &lt;210> 32 &lt;211&gt; 72 &lt;212&gt; DNA &lt;213&gt; artificial sequence

&lt;220> &lt;223>optUV 1:2 signal peptide-IL-2 &lt;400&gt; 32 atgtatagaa tgcagctcct gtcctgcatt gccctgagcc tcgccctcgt gacaaactcc gcccctacct cc 60 72 &lt;210> 33 &lt;211&gt; 22S &lt;212&gt; Wk &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; wtUV signal peptide - human TMMTNF superfamily, member 2) &lt; 400 &gt; 33 atgagcactg aaagcatgat ccgggacgtg gagctggccg aggaagccct ccccaagaaa accggcggcc cccaggggag cagaagatgt ttgttcctga gcctgttctc cttcctgatc gtggcaggcg ctaccaccct gttctgcctg ctgcactttg gagtgatcgg cccccagagg gaggagttcc ccagggacct ctctctaatc Agccctctgg cccaggca 60 120 180 &lt;210> 34 &lt;211&gt; 228 &lt;212&gt; mk • ΊΊ · 228 201207108 &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; optUV 1:2 signal peptide - human tumor necrosis factor (TOF superfamily, member 2) &lt;400&gt; 34 atgtccaccg aaagcatgat ccgggacgtg gagctggccg aggaagccct gcctaagaaa 60 accggaggcc ctcagggaag caggagatgt ctgtttctgt ccctgtttag ctttctgatt 120 gtggctggcg ctaccacact gttttgcctc ctgcatttcg gagtgattgg ccctcagagg 180 gaggagttcc ctagagacct gtccctga Tt agccctctgg ctcaggct 228 &lt;210> 35 &lt;211&gt; 474 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; wtUV numbering sequence human TOF &lt;TNF superfamily, member 2) &lt;400&gt;; 35 gtcagatcat cttctcgaac cccgagtgac aagcctgtag cccatgttgt agcaaaccct 60 caagccgagg gccagctcca gtggctgaac cgccgggcca atgccctgct cgccaacggc 120 gtcgagctga gagataacca gctggtggtg ccatcagagg gcctgtacct catctactcc 180 caggtcctgt tcaagggcca aggctgcccc tccacccatg tgctcctcac ccacaccatc 240 agccgcatcg ccgtgagcta ccagaccaag gtcaacctcc tctctgccat caagagcccc 300 tgccagaggg agaccccaga gggggccgag gccaagccct ggtatgagcc catctacctc 360 ggcggggtgt tccagctgga gaagggtgac cgactcagcg ctgagatcaa tagacccgac 420 tatctcgact ttgccgagag cggccaggtg tactttggga tcattgccct gtga 474 &lt;210> 36 &lt;211&gt; 474 &lt;212&gt; DNA &lt;213&gt;Artificial sequence&lt;223&gt; optUV 1:2 coding sequence Homo sapiens tumor necrosis factor (ΊΜ7 Superfamily, member 2) &lt;220&gt;&lt;400> 36 gtgagaagca gcagcaggac ccctagcgat aagcctgtgg ctcacgtcgt cgctaac cct 60 caggccgagg gccagctcca gtggctgaat agaagggcca atgccctgct cgccaacggc 120 gtcgagctga gagacaatca gctcgtggtc ccctccgagg gactgtatct gatttactcc 180 caggtcctgt ttaagggaca gggatgccct agcacacacg tcctgctgac ccacaccatt 240 agcaggatcg ctgtgtccta ccaaaccaaa gtgaatctgc tgtccgctat caaaagccct 300 tgccaaagag aaacccctga gggagccgaa gccaaaccct ggtacgaacc catttacctc 360 ggcggagtgt ttcagctgga gaaaggcgat agactcagcg ctgagattaa caggcccgat 420 tacctcgact ttgccgaaag cggacaggtc tactttggca ttatcgctct gtaa 474 &lt;210&gt; 37 &lt;211&gt; 157 &lt;212&gt; PRT &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Amino acid sequence of TNF-α 201207108 &lt;400&gt;

Val Arg Ser Ser Ser Arg Thr Pro Ser Asp Lys Pro Val Ala His Val 15 l〇 15

Val Ala Asn Pro Gin Ala GIu Gly Gin Leu Gin Trp Leu Asn Arg Arg 20 25 30

Ala Asn Ala Leu Leu Ala Asn Gly Val Glu Leu Arg Asp Asn Gin Leu 35 40 45

Val Val Pro Ser Glu Gly Leu Tyr Leu lie Tyr Ser Gin Val Leu Phe 50 55 60

Lys Gly Gin Gly Cys Pro Ser Thr His Val Leu Leu Thr His Thr lie 65 70 75 80

Ser Arg lie Ala Val Ser Tyr Gin Thr Lys Val Asn Leu Leu Ser Ala 85 90 95

Lie Lys Ser Pro Cys Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys 100 105 110

Pro Trp Tyr Glu Pro lie Tyr Leu Gly Gly Val Phe Gin Leu Glu Lys 115 120 125

Gly Asp Arg Leu Ser Ala Glu lie Asn Arg Pro Asp Tyr Leu Asp Phe 130 135 140

Ala Glu Ser Gly Gin Val Tyr Phe Gly lie lie Ala Leu 145 150 155 &lt;210&gt; 38 &lt;211&gt; 132 &lt;212&gt; DNA &lt;213&gt;Artificial Sequence&lt;220&gt;&lt;223&gt; SV40e 3Reg Contains PolyA &lt;;400&gt; 38 60 120 132 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgt£gtttgt ccaaactcat caatgtatct tatcatgtct gg &lt;210> 39 &lt;211&gt; 627 &lt;212&gt; DNA &lt;213&gt;Artificial Sequence&lt;220&gt;&lt; 223 &gt; hGH 3Reg containing PolyA &lt; 400 &gt; 39 60 120 180 tgatgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc 240 .79 - 201207108 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga 300 Gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag 360 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc 420 accttggcct ccca aattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct 480 tctgatttta aaataactat accagcagga ggacgtccag acacagcata ggctacctgg 540 ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg 600 gtccactcag tagatgcctg ttgaatt 627 &lt; 210 &gt; 40 &lt; 211 &gt; 1155 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; al WF a 3'UTR &lt; 400 &gt; 40 ggaggacgaa catccaacct tcccaaacgc ctcccctgcc ccaatccctt tattaccccc 60 tccttcagac accctcaacc tcttctggct caaaaagaga attgggggct tagggtcgga 120 acccaagctt agaactttaa gcaacaagac caccacttcg aaacctggga ttcaggaatg 180 tgtggcctgc acagtgaagt gctggcaacc actaagaatt caaactgggg cctccagaac 240 tcactggggc ctacagcttt gatccctgac atctggaatc tggagaccag ggagcctttg 300 gttctggcca gaatgctgca ggacttgaga agacctcacc tagaaattga cacaagtgga 360 ccttaggcct tcctctctcc agatgtttcc agacttcctt gagacacgga gcccagccct 420 ccccatggag ccagctccct ctatttatgt ttgcacttgt gattatttat tatttattta 480 ttatttattt atttacagat gaatgtattt atttgggaga ccggggtatc ctgggggacc 540 caatgtagga gctgc cttgg ctcagacatg 111tccgtga aaacggagct gaacaatagg 600 ctgttcccat gtagccccct ggcctctgtg ccttcttttg attatgtttt ttaaaatatt 660 tatctgatta agttgtctaa acaatgctga tttggtgacc aactgtcact cattgctgag 720 cctctgctcc ccaggggagt tgtgtctgta atcgccctac tattcagtgg cgagaaataa 780 agtttgctta gaaaagaaac atggtctcct tcttggaatt aattctgcat ctgcctcttc 840 ttgtgggtgg gaagaagctc cctaagtcct ctctccacag gctttaagat ccctcggacc 900 cagtcccatc cttagactcc tagggccctg gagaccctac ataaacaaag cccaacagaa 960 tattccccat cccccaggaa acaagagcct gaacctaatt acctctccct cagggcatgg 1020 gaatttccaa ctctgggaat tccaatcctt gctgggaaaa tcctgcagct caggtgagat 1080 ttccggctgt tgcagctggc cagcagtccg gagagagctg gagaggagcc gcattctcag 1140 gtacctgaat cacac 1155 &lt; 210 &gt; 41 &lt; 211 &gt; 1155 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 & gt Human TNF a: 3'UTR AtoC mutation &lt;400&gt; 41 ggaggacgaa catccaacct tcccaaacgc ctcccctgcc ccaatccctt tattaccccc 60 -80 120 120

201207108 tccttcagac accctcaacc tcttctggct caaaaagaga attgggggct tagggtcgga acccaagctt agaactttaa gcaacaagac caccacttcg aaacctggga ttcaggaatg tgtggcctgc acagtgaagt gctggcaacc actaagaatt caaactgggg cctccagaac tcactggggc ctacagcttt gatccctgac atctggaatc tggagaccag ggagcctttg gttctggcca gaatgctgca ggacttgaga agacctcacc tagaaattga cacaagtgga ccttaggcct tcctctctcc agatgtttcc agacttcctt gagacacgga gcccagccct ccccatggag ccagctccct ctatttatgt ttgcacttgt gattctttct tctttctttc ctttacagat gaatgtattt atttgggaga ccggggtatc ctgggggacc caatgtagga ttctttcttt gctgccttgg ctcagacatg ttttccgtga aaacggagct gaacaatagg ctgttcccat gtagccccct ggcctctgtg ccttcttttg attatgtttt ttaaaatatt tatctgatta agttgtctaa acaatgctga tttggtgacc aactgtcact cattgctgag cctctgctcc ccaggggagt tgtgtctgta atcgccctac tattcagtgg cgagaaataa agtttgctta gaaaagaaac atggtctcct tcttggaatt aattctgcat ctgcctcttc ttgtgggtgg gaagaagctc cctaagtcct ctctccacag gctttaagat ccctcggacc cagtcccatc cttagactcc tagggccctg gagaccctac ataaacaaag cccaacagaa Tattccccat cccccaggaa acaagagcct gaacctaatt acctctccct cagggcatgg gaatttccaa ctctgggaat tccaatcctt gctgggaaaa tcctgcagct caggtgagat ttccggctgt tgcagctggc cagcagtccg gagagagctg gagaggagcc gcattctcag gtacctgaat cacac

&lt;210&gt; 42 &lt;211&gt; 618 &lt;212&gt; DNA &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; Human GAST, 3_-UTR &lt;400&gt; 42 caatcctaga accaagcttc agagcctagc cacctcccac cccactccag ccctgtcccc tgaaaaactg atcaaaaata aactagtttc cagtggatca atggactgtg tcagtgttgt agggcagagg agggggactc atctgggctc atctgggggt gaagttgtgg cagggactaa gagctgagtg cctcttaggg gcagggaccg tcccccagag ccccacattg aacgagaatc cacaggtatg ggcaggataa tatatggtag ggttcatagc cagagtaacc ttttttttta atttttattt tattttattt ttgagatgga gtttcgctct tgtctcccag gctggagtgc aataatgaga cctcagctca ctgcaacctc tgcctcctag gttcaagcga ttttcctgcc tcagcctccc aagtagctgg gattacaggt gcccgccacc acacctggct aatttttttg tatttttagt ggggacgggg tttcaccatg ttggccaggc tggtcttgaa ctcctgacct cagtgatcca cccgcctcgg cctcccaaag tgctgggatt acagcatgag ccaccgtgcc Cagcctcaga gtaagctt

&lt;210> 43 &lt;211&gt; 589 &lt;212&gt; DNA 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1155 60 120 180 240 300 360 420 480 540 600 618 -81 - 60 201207108 &lt;; 213 &gt; artificial sequence &lt; &lt;; 220 & gt 223 &gt;synthetic; T REG &lt; 400 &gt; 43 120 180 240 300 360 420 480 540 589 ttctcaagaa agccctcatt tttataacct ggcaaaatct tgttaatgtc attgctaaaa aataaataaa agctagatac tggaaaccta actgcaatgt ggatgtttta cccacatgac ttattatgca taaagccaaa tttccagttt aagtaattgc ctacaataaa aagaaatttt gcctgccatt ttcagaatca tcttttgaag ctttctgttg atgttaactg agctactaga gatattct'ta tttcactaaa tgtaaaattt ggagtaaata tatatgtcaa tatttagtaa agcttttctt ttttaatttc caggaaaaaa taaaaagagt atgagtcttc tgtaattcat tgagcagtta gctcatttga gataaagtca aatgccaaac actagctctg tattaatccc catcattact ggaataaaag atctttattt tcattagatc tgtgtgttgg ttttttgtgt gggccttggg ggagggggag gccagaatgg ccttggggga gggggaggcc agaatggcct tgggggaggg ggaggccaga atggccttgg gggaggggga ggccagaat &lt; 210 &gt; 44 &lt;211&gt; 102 &lt;212&gt; DNA &lt;2 13&gt;Artificial sequence&lt;220&gt;&lt;223&gt; Human GAPDH 5UTR &lt;400&gt; 44 60 102 aaattgagcc cgcagcctcc cgcttcgctc tctgctcctc ctgttcgaca gtcagccgca tcttcttttg cgtcgccagc cgagccacat cgctcagaca cc &lt;210&gt; 45 &lt;211&gt; 72 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; wtUV insulin SP coding sequence &lt;400&gt; 45 60 72 atggccctgt ggatgcgcct cctgcccctg ctggcgctgc tggccctctg gggacctgac ccagccgcag cc &lt;210&gt; 46 &lt;211&gt; 96 &lt;212> DNA &lt;213&gt; Artificial sequence &lt;220&gt; _ &lt;223&gt; wtUV human FGF-19 signal peptide &lt;400> 46 60 atgcggagcg ggtgtgtggt ggtccacgta tggatcttgg ccggactgtg gctggccgtg gccgggagac ccctcgcctt ctcggacgcc ggaccc &lt;210&gt; 47 &lt;211&gt; 10780 &lt;212&gt; DNA &lt;213&gt;Artificial sequence -82 - &lt;220> 96 60 60

201207108 &lt; 223> PoIyDNA_from a WN_43318 &lt; 400 &gt; 47 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgiggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggag ttgtgtgaat actc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca catcccctga caagctgcca ggcaggttct cttcctctca catactgacc cacggctcca ccctctctcc cctggaaagg acaccgctag cgccaccatg agcactgaaa gcatgatccg ggacgtggag ctggccgagg aagccctccc caagaaaacc ggcggccccc aggggagcag aagatgtttg ttcctgagcc tgttttcctt cctgatcgtg gcaggcgcta ccaccctgtt ctgcctgctg cactttggag tgatcggccc ccagagggag gagttcccca gggacctctc tctaatcagc cctctggccc aggcaggatc cgtcagatca tcttctcgaa ccccgagtga caagcctgta gcccatgttg tagcaaaccc tcaagccgag ggccagctcc agtggctgaa ccgccgggcc aatgccctgc tcgccaacgg cgtcgagctg agagataacc agctggtggt gccatcagag ggcctgtacc tcatctactc ccaggtcctg ttcaagggcc aaggctgccc ctccacccat gtgctcctca cccacaccat cagccgcatc gccgtgagct accagaccaa ggtcaacctc ctctctgcca tcaagagccc ctgccagagg gagaccccag agggggccga ggccaagccc tggtatgagc ccatc tacct cggcggggtg ttccagctgg agaagggtga ccgactcagc gctgagatca atagacccga ctatctcgac tttgccgaga gcggccaggt gtactttggg atcattgccc tgtgaatcga ttcgtacgtc gacatcgaga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg ggcgcgccgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 10S0 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 -83 - 201207108 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga 2220 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac 2280 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga 2340 Acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg 2400 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct 2460 gctgggctgg gtacgtgcgc tcggggt tgg cgagtgtgtt ttgtgaagtt ttttaggcac 2520 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt 2580 ccgctaaatt ctggccgttt ttggcttttt tgttagacga gctagcgccg ccaccatggg 2640 ccctaaaaag aagcgtaaag tcgccccccc gaccgatgtc agcctggggg acgagctcca 2700 cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga 2760 catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta 2820 cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat 2880 tgacgagtac ggtggggaat tcgagatgcc tgtggacagg atcctggagg cagagcttgc 2940 tgtggaacag aagagtgacc agggcgttga gggtcctggg ggaaccgggg gtagcggcag 3000 cagcccaaat gaccctgtga ctaacatctg tcaggcagct gacaaacagc tattcacgct 3060 tgttgagtgg gcgaagagga tcccacactt ttcctccttg cctctggatg atcaggtcat 3120 attgctgcgg gcaggctgga atgaactcct cattgcctcc ttttcacacc gatccattga 3180 tgttcgagat ggcatcctcc ttgccacagg tcttcacgtg caccgcaact cagcccattc 3240 agcaggagta ggagccatct ttgatcgggt gctgacagag ctagtgtcca aaatgcgtga 3300 catgaggatg gacaagacag agcttggctg cctgagggca atcattctgt ttaatccaga 3360 ggtgaggggt ttgaaatccg cccaggaagt tgaacttcta cgtgaaaaag tatatgccgc 3420 tttggaagaa tatactagaa caacacatcc cgatgaacca ggaagatttg caaaactttt 3480 gcttcgtctg ccttctttac gttccatagg ccttaagtgt ttggagcatt tgtttttctt 3540 tcgcct tat t ggagatgttc caattgatac gttcctgatg gagatgcttg aatcaccttc 3600 tgat tcataa tctagcctag cccccctctc cctccccccc ccctaacgtt actggccgaa 3660 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atat tgccgt 3720 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 3780 gtct t tcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 3840 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 3900 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 3960 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 4020 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 4080 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg Ttaaaaaacg 4140 tctaggc ccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat ctctaggcgc 4200 caccatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac t taaaaagct 4260 caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg 4320 ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc 4380 aaggctagaa agactggaac agctatttct actgattttt cctcgagaag acct tgacat 4440 -84 -

201207108 gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcgccgga attcccgggg atccggcctg agtgcgtagt acccgagact cagtgcgcca tgaagcggaa agagaagaaa gcacagaagg agaaggacaa actgcctgtc agcacgacga cggtggacga ccacatgccg cccattatgc agtgtgaacc tccacctcct gaagcagcaa ggattcacga agtggtccca aggtttctct ccgacaagct gttggtgaca aaccggcaga aaaacatccc ccagttgaca gccaaccagc agttccttat cgccaggctc atctggtacc aggacgggta cgagcagcct tctgatgaag atttgaagag gattacgcag acgtggcagc aagcggacga tgaaaacgaa gagtcggaca ctcccttccg ccagatcaca gagatgacta tcctcacggt ccaacttatc gtggagttcg cgaagggatt gccagggitc gccaagatct cgcagcctga tcaaattacg ctgcttaagg cttgctcaag tgaggtaatg atgctccgag tcgcgcgacg atacgatgcg gcctcagaca gtattctgtt cgcgaacaac caagcgtaca ctcgcgacaa ctaccgcaag gctggcatgg ccgaggtcat cgaggatcta ctgcacttct gccggtgcat gtactctatg gcgttggaca acatccatta cgcgctgctc acggctgtcg tcatcttttc tgaccggcca gggttggagc agccgcaact ggtggaagag atccagcggt actacctgaa tacgctccgc atctatatcc tgaaccagct gagcgggtcg gcgcgttcgt ccgtcatata cggcaagatc ctctcaatcc tctctgagct acgcacgctc ggcatgcaaa actccaacat gtgcatctcc ctcaagctca agaacagaaa gctgccgcct ttcctcgagg agatctggga tgtggcggac atgtcgcaca cccaaccgcc gcctatcctc gagtccccca cgaatctcta ggcggcctct agagcggccg ccaccgcggg gagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgagg tcgactctag tccccgcggt ggcagatctg gaaggtgctg aggtacgatg agacccgcac caggtgcaga ccctgcgagt gtggcggtaa acatattagg aaccagcctg tgatgctgga tgtgaccgag gagctgaggc ccgatcactt ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat gaagatacag attgaggtac tgaaatgtgt gggcgtggct taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc cgcgggattg tgactgactt tgctttcctg agcccgcttg 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 -85 - 201207108 caagcagtgc agcttcccgt tcatccgccc gcgatgacaa gttgacggct cttttggcac 6720 aattggattc tttgacccgg gaacttaatg tcgtttctca gcagctgttg gatctgcgcc 6780 agcaggtttc tgccctgaag gcttcctccc ctcccaatgc Ggtttaaaac ataaataaaa 6840 aaccagactc tgtttggatt tggatcaagc aagtgtcttg ctgtctttat ttaggggttt 6900 tgcgcgcgcg gtaggcccgg gaccagcggt ctcggtcgtt gagggtcctg tgta tttttt 6960 ccaggacgtg gtaaaggtga ctctggatgt tcagatacat gggcataagc ccgtctctgg 7020 ggtggaggta gcaccactgc agagcttcat gctgcggggt ggtgttgtag atgatccagt 7080 cgtagcagga gcgctgggcg tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca 7140 ggggcaggcc cttggtgtaa gtgtttacaa agcggttaag ctgggatggg tgcatacgtg 7200 gggatatgag atgcatcttg gactgtattt ttaggttggc tatgttccca gccatatccc 7260 tccggggatt catgttgtgc agaaccacca gcacagtgta tccggtgcac ttgggaaatt 7320 tgtcatgtag cttagaagga aatgcgtgga agaacttgga gacgcccttg tgacctccaa 7380 gattttccat gcattcgtcc ataatgatgg caatgggccc acgggcggcg gcctgggcga 7440 agatatttct gggatcacta acgtcatagt tgtgttccag gatgagatcg tcataggcca 7500 tttttacaaa gcgcgggcgg agggtgccag actgcggtat aatggttcca tccggcccag 7560 gggcgtagtt accctcacag atttgcattt cccacgcttt gagttcagat ggggSgatca 7620 tgtctacctg cggggcgatg aagaaaacgg tttccggggt aggggagatc agctgggaag 7680 aaagcaggtt cctgagcagc tgcgacttac cgcagccggt gggcccgtaa atcacaccta 7740 ttaccggctg caactggtag ttaagagagc tgcagctgcc gtcatcc ctg agcagggggg 7800 ccacttcgtt aagcatgtcc ctgactcgca tgttttccct gaccaaatcc gccagaaggc 7860 gctcgccgcc cagcgatagc agttcttgca aggaagcaaa gtttttcaac ggtttgagac 7920 cgtccgccgt aggcatgctt ttgagcgttt gaccaagcag ttccaggcgg tcccacagct 7980 cggtcacctg ctctacggca tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg 8040 gctttcgctg tacggcagta gtcggtgctc gtccagacgg gccagggtca tgtctttcca 8300 cgggcgcagg gtcctcgtca gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg 8160 cgcgctggcc agggtgcgct tgaggctggt cctgctggtg ctgaagcgct gccggtcttc 8220 gccctgcgcg tcggccaggt agcatttgac catggtgtca tagtccagcc cctccgcggc 8280 gtggcccttg gcgcgcagct tgcccttgga ggaggcgccg cacgaggggc agtgcagact 8340 tttgagggcg tagagcttgg gcgcgagaaa taccgattcc ggggagtagg catccgcgcc 8400 gcaggccccg cagacggtct cgcattccac gagccaggtg agctctggcc gttcggggtc 8460 aaaaaccagg tttcccccat gctttttgat gcgtttctta cctctggttt ccatgagccg 8520 gtgtccacgc tcggtgacga aaaggctgtc cgtgtccccg tatacagact tgagaggcct 8580 gtcctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg 8640 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg 8700 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga 8760 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg 8820 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg 8880 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga 8940 -86 - 201207108 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tc ttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat (:, ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtg Agta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt

t I \ ^ J gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa &lt; 210 &gt; 48 &lt; 211 &gt; 10780 &lt;212&gt; I^A &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; PolyDNA_from_WN-43319 &lt;400&gt; 48 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 9720 9780 9840 9900 9960 10020 10080 10140 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10780 -87 - 201207108 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa 180 tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag 240 tgtggcggaa gtgtgatgt t gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa 300 aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta 360 ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa 420 actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg 480 tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 780 tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg 1020 accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca 1080 catcccctga caagctgcca gg caggttct cttcctctca catactgacc cacggctcca 1140 ccctctctcc cctggaaagg acaccgctag cgccaccatg tccaccgaaa gcatgatccg 1200 ggacgtggag ctggccgagg aagccctgcc taagaaaacc ggaggccctc agggaagcag 1260 gagatgtctg tttctgtccc tgtttagctt tctgattgtg gctggcgcta ccacactgtt 1320 ttgcctcctg catttcggag tgattggccc tcagagggag gagttcccta gagacctgtc 1380 cctgattagc cctctggctc aggctggatc cgtgagaagc agcagcagga cccctagcga 1440 taagcctgtg gctcacgtcg tcgctaaccc tcaggccgag ggccagctcc agtggctgaa 1500 tagaagggcc aatgccctgc tcgccaacgg cgtcgagctg agagacaatc agctcgtggt 1560 cccctccgag ggactgtatc tgatttactc ccaggtcctg tttaagggac agggatgccc 1620 tagcacacac gtcctgctga cccacaccat tagcaggatc gctgtgtcct accaaaccaa 1680 agtgaatctg ctgtccgcta tcaaaagccc ttgccaaaga gaaacccctg agggagccga 1740 agccaaaccc tggtacgaac ccat ttacct cggcggagtg tttcagctgg agaaaggcga 1800 tagactcagc gctgagatta acaggcccga t tacctcgac tttgccgaaa gcggacaggt I860 ctactttggc attatcgctc tgtaaatcga ttcgtacgtc gacatcgaga acttgtttat 1920 tgcagcttat aat ggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 1980 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 2040 ggcgcgccgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc 2100 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct 2160 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga 2220 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac 2280 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt -88 - ggggcggtga 2340

201207108 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg gtacgtgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga gctagcgccg ccaccatggg ccctaaaaag aagcgtaaag tcgccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtggggaat tcgagatgcc tgtggacagg atcctggagg cagagcttgc tgtggaacag aagagtgacc agggcgttga gggtcctggg ggaaccgggg gtagcggcag cagcccaaat gaccctgtga ctaacatctg tcaggcagct gacaaacagc tattcacgct tgttgagtgg gcgaagagga tcccacactt ttcctccttg cctctggatg atcaggtcat attgctgcgg gcaggctgga atgaactcct cattgcctcc ttttcacacc gatccattga tgttcgagat ggcatcctcc ttgccacagg tcttcacgtg caccgcaact cagcccattc agcaggagta ggagccatct ttgatcgggt gctgacagag ctagtgtcca aaatgcgtga catgaggatg gacaagacag agcttggctg cctgagggca atcattctgt ttaatccaga ggtgaggggt ttgaaatccg cccaggaagt tgaacttcta cgtgaaaaag tatatgccgc tttggaagaa tatactagaa caacacatcc cgatgaacca ggaagatttg caaaactttt gcttcgtctg ccttctttac gttccatagg ccttaagtgt ttggagcatt tgtttttctt tcgccttatt ggagatgttc caattgatac gttcctgatg gagatgcttg aatcaccttc tgattcataa tctagcctag cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat ctctaggcgc caccatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 -89 - 201207108 tctaacattg agacagcata gaataagtgc gacatcatca teggaagaga gtagtaacaa 4620 aggtcaaaga cagttgactg tatcgccgga attcccgggg atccggcctg agtgcgtagt 4680 acccgagact cagtgcgcca tgaagcggaa agagaagaaa Gcacagaagg agaaggacaa 4740 actgcctgtc agcacgacga cggtggacga ccacatgccg cccattatgc agtgtgaacc 4800 tccacctcct gaagcagcaa ggattcacga agtggtccca aggtttctct ccga caagct 4860 gttggtgaca aaccggcaga aaaacatccc ccagttgaca gccaaccagc agttccttat 4920 cgccaggctc atctggtacc aggacgggta cgagcagcct tetgatgaag atttgaagag 4980 gattacgcag acgtggcagc aagcggacga tgaaaacgaa gagtcggaca ctcccttccg 5040 ccagatcaca gagatgacta tcctcacggt ccaacttatc gtggagttcg egaagggatt 5100 gccagggttc gccaagatct cgcagcctga tcaaattacg ctgcttaagg cttgctcaag 5160 tgaggtaatg atgctccgag tcgcgcgacg atacgatgcg gcctcagaca gtattctgtt 5220 cgcgaacaac caagcgtaca ctcgcgacaa ctaccgcaag gctggcatgg ccgaggtcat 5280 cgaggatcta ctgcacttct gccggtgcat gtactctatg gcgttggaca acatccatta 5340 cgcgctgctc acggctgtcg tcatcttttc tgaccggcca agccgcaact gggttggagc gcctatcctc gagtccccca cgaatctcta 5400 ggtggaagag atccagcggt actacctgaa tacgctccgc atctatatcc tgaaccagct 5460 gagcgggtcg gcgcgttcgt ccgtcatata cggcaagatc ctctcaatcc tetetgaget 5520 acgcacgctc ggcatgcaaa actccaacat gtgcatctcc ctcaagctca agaacagaaa 5580 gctgccgcct ttcctcgagg agatctggga tgtggcggac atgtcgcaca cccaaccgcc 5640 ggcggcctct agagegg eeg ccaccgcggg 5700 gagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgeagt 5760 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa 5820 gctgcaataa acaagttaac aacaacaat t gcat teat 11 tatgtttcag gttcaggggg 5880 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg 5940 atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 6000 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 6060 gtcagcgggt gttggcgggt gtcggggcgc agccatgagg tcgactctag tccccgcggt 6120 gccggggtgc 6480 gtcagaatgt ggcagatctg gaaggtgctg aggtacgatg agacccgcac caggtgcaga ccctgcgagt 6180 gtggcggtaa acatattagg aaccagcctg tgatgctgga tgtgaccgag gagetgagge 6240 ccgatcactt ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat gaagatacag 6300 attgaggtac tgaaatgtgt gggcgtggct taagggtggg aaagaatata taaggtgggg 6360 gtcttatgta gttttgtatc tgttttgcag cagccgccgc cgccatgagc accaactcgt 6420 ttgatggaag cattgtgagc tcatatttga caacgcgcat gcccccatgg gatgggctcc agcattgatg Gtcgccc cgt cctgcccgca aactctacta 6540 ccttgaccta cgagaccgtg tctggaacgc cgttggagac tgcagcctcc gccgccgct t 6600 cagccgctgc agccaccgcc cgcgggattg tgactgactt tgctttcctg agcccgcttg 6660 caagcagtgc agcttcccgt tcatccgccc gcgatgacaa gttgaegget cttttggcac 6720 aattggattc tttgacccgg gaacttaatg tcgtttctca gcagctgttg gatctgcgcc 6780 agcaggtttc tgccctgaag gcttcctccc ctcccaatgc ggt t taaaac -90 - ataaataaaa 6840 201207108

aaccagactc tgtttggatt tggatcaagc aagtgtcttg ctgtctttat ttaggggttt 6900 tgcgcgcgcg gtaggcccgg gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt 6960 ccaggacgtg gtaaaggtga ctctggatgt tcagatacat gggcataagc ccgtctctgg 7020 ggtggaggta gcaccactgc agagcttcat gctgcggggt ggtgttgtag atgatccagt 7080 cgtagcagga gcgctgggcg tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca 7140 ggggcaggcc cttggtgtaa gtgtttacaa agcggttaag ctgggatggg tgcatacgtg 7200 gggatatgag atgcatcttg gactgtattt ttaggttggc tatgttccca gccatatccc 7260 tccggggatt catgttgtgc agaaccacca gcacagtgta tccggtgcac ttgggaaatt 7320 tgtcatgtag cttagaagga aatgcgtgga agaacttgga gacgcccttg tgacciccaa 7380 gattttccat gcattcgtcc ataatgatgg caatgggccc acgggcggcg gcctgggcga 7440 agatatttct gggatcacta acgtcatagt tgtgttccag gatgagatcg tcataggcca 7500 tttttacaaa gcgcgggcgg agggtgccag actgcggtat aatggttcca tccggcccag 7560 gggcgtagtt accctcacag atttgcattt cccacgcttt gagttcagat ggggggatca 7620 tgtctacctg cggggcgatg aagaaaacgg tttccggggt aggggagatc agcigggaa g 7680 aaagcaggtt cctgagcagc tgcgacttac cgcagccggt gggcccgtaa atcacaccta 7740 ttaccggctg caactggtag ttaagagagc tgcagctgcc gtcatccctg agcagggggg 7800 ccacttcgtt aagcatgtcc ctgactcgca tgttttccct gaccaaatcc gccagaaggc 7860 gctcgccgcc cagcgatagc agttcttgca aggaagcaaa gtttttcaac ggtttgagac 7920 cgtccgccgt aggcatgctt ttgagcgttt gaccaagcag ttccaggcgg tcccacagct 7980 cggtcacctg ctctacggca tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg 8040 gctttcgctg tacggcagta gtcggtgctc gtccagacgg gccagggtca tgtctttcca 8100 cgggcgcagg gtcctcgtca gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg 8160 cgcgctggcc agggtgcgct tgaggctggt cctgctggtg ctgaagcgct gccggtcttc 8220 gccctgcgcg tcggccaggt agcatttgac catggtgtca tagtccagcc cctccgcggc 8280 gtggcccttg gcgcgcagct tgcccttgga ggaggcgccg cacgaggggc agtgcagact 8340 tttgagggcg tagagcttgg gcgcgagaaa taccgattcc ggggagtagg catccgcgcc 8400 gcaggccccg cagacggtct cgcattccac gagccaggtg agctctggcc gttcggggtc 8460 aaaaaccagg tttcccccat gctttttgat gcgtttctta cctctggttt c catgagccg 8520 gtgtccacgc tcggtgacga aaaggctgtc cgtgtccccg tatacagact tgagaggcct 8580 gtcctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg 8640 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg 8700 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga 8760 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg 8820 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg 8880 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga 8940 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg 9000 tagatgacga ccatcaggga cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg 9060 201207108 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccg ct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctcctt cgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata Tttgaatgta tttagaaaaa &lt;210&gt; 49 &lt;211&gt; 10624 &lt;212&gt; mk &lt; 213 &gt; artificial sequence &lt;220&gt;&lt;223&gt; PolyDNA_from_WN-43320 &lt;400&gt; 49 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct Ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cglggcgcgg ggcgtgggaa cggggcgggt gacgtagtag -92 - 9120 9180 9240 9300 9360 9420 94 80 9540 9600 9660 9720 9780 9840 9900 9960 10020 10080 10140 10200 10260 10320 30380 10440 10500 10560 10620 10680 10740 10780 60 120 180 240 201207108

tgtggcggaa aagtgacgtt ggcggatgtt actgaataag tctagggaga tgagtattag tgttggtttt acaaaacaaa ctatcgataa tactgtcctc tgtcctccga ttccttcgaa ctggtgtgtg accagctaag catcccctga ccctctctcc ctgcattgcc aagcagcagc cgagggccag gctgagagac cctgtttaag gatcgctgtg aagagaaacc agtgtttcag cgactttgcc cgtcgacatc cacaaatttc catcaatgta cgcgggcgcc tcctgatcct agaaccccag ggttttcttt ggagggatct acagctagtt tcgtcacttg tgttttgtga ttcagtgtta gtgtgatgtt tttggtgtgc gtagtaaatt aggaagtgaa tccggtaccg gcgcgcacca ttgtgtgaat ctagcaaaat tgcaggtcgg cgagcggagt gcggagactc ggaagagggg agctcatctt agggagagaa caagctgcca cctggaaagg ctgagcctcg aggaccccta ctccagtggc aatcagctcg ggacagggat tcctaccaaa cctgagggag ctggagaaag gaaagcggac gagaacttgt acaaataaag tcttatcatg cccctcctca tccgcccgga tatcagcaga ccagagagcg ccgtggggcg ccgtcgcagc gtgagtagcg agttttttag gactagtaaa gcaagtgtgg gccggtgtac tgggcgtaac atctgaataa gcgcgcgcgc ggtgccgcaa cgatagtact aggctgtccc agtactgtcc actgtcctcc ttcgaaggaa cggggtcgaa cctgtagatc gcaactacag ggcaggttct acaccgctag ccctcgtgac gcgataagcc tgaatagaag tggtcccctc gccctagcac ccaaagtgaa ccgaagccaa gcgatagact aggtctactt ttattgcagc catttttttc tctgggcgcg cggcgagcgc cgctcaggac aggacatttt gaacaggcga gtgaacgccg cgggatttgg ggctgctggg gcaccttttg ttgtccgcta cggaacacat acaggaagtg cgagtaagat ttttgtgtta cgtttggccg taaaatatct aacatacgct cagtgcaagt tccgagcgga gagcggagta gaggggcggg gacctagagg acgcgtctcc accccccctg cttcctctca cgccaccatg aaactccgcc tgtggctcac ggccaatgcc cgagggactg acacgtcctg tctgctgtcc accctggtac cagcgctgag tggcattatc ttataatggt actgcattct ccggcctccg tgccacgtca agcggcccgc aggacgggac ggaaaagtag atgattatat gtcgcggttc ctgggtacgt aaatgtaatc aattctggcc gtaagcgacg acaattttcg ttggccattt ctcatagcgc cctcgagtct ttattttcat ctccatcaaa gcaggtgcca gtactgtcct ctgtcctccg gtcgatcgac gtatataatg ctcagcaagg aaaacaaccc catactgacc tatagaatgc cctaccagcg gtcgtcgcta ctgctcgcca tatctgattt ctgacccaca gctatcaaaa gaacccattt attaacaggc gctctgtaaa tacaaataaa agttgtggu cgccgggttt gacgaagggc tgctcataag ttgggtgact tcccttctcg aaggacgcgc ttgtttgtgg gcgctcgggg atttgggtca gtttttggct gatgtggcaa cgcggtttta tcgcgggaaa gtaatatttg agagatccgg tacatctgtg acaaaacgaa gaacatttct ccgagcggag agcggagtac cccgcccctc ggtgccttag acagcagagg tcagacgcca cacggctcca agctcctgtc gatccgtgag accctcaggc acggcgtcga actcccaggt ccattagcag gcccttgcca acctcggcgg ccgattacct tcgattcgta gcaatagcat tgtccaaact tggcgcctcc ttggcgagtg atatgtaatt gcagcgagcg actcggcctt ctagggcact gcgattctgc Cgggtgtggc atcgctgtga tttttgttag 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 -93 - 201207108 acgagctagc gccgccacca tgggccctaa aaagaagcgt aaagtcgccc ccccgaccga 2520 tgtcagcctg ggggacgagc tccacttaga cggcgaggac gtggcgatgg cgcatgccga 2580 cgcgctagac gatttcgatc tggacatgtt gggggacggg gattccccgg Gtccgggatt 2640 taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca 2700 gatgtttacc gatgcccttg gaattgacga gtacggtggg gaattcgaga tgcctgtgga 2760 caggatcctg gaggcagagc ttgctgtgga acagaagagt gaccagggcg ttgagggtcc 2820 tgggggaacc gggggtagcg gcagcagccc aaatgaccct gtgactaaca tctgtcaggc 2880 agctgacaaa cagctattca cgcttgttga gtgggcgaag aggatcccac acttttcctc 2940 cttgcctctg gatgatcagg tcatattgct gcgggcaggc tggaatgaac tcctcattgc 3000 ctccttttca caccgatcca ttgatgttcg agatggcatc ctccttgcca caggtcttca 3060 cgtgcaccgc aactcagccc attcagcagg agtaggagcc atctttgatc gggtgctgac 3120 agagctagtg tccaaaatgc gtgacatgag gatggacaag acagagcttg gctgcctgag 3180 ggcaatcatt ctgtttaatc cagaggtgag gggtttgaaa tccgcccagg aagttgaact 3240 tctacgtgaa aaagtatatg ccgctttgga agaatatact agaacaacac atcccgatga 3300 accaggaaga tttgcaaaac ttttgcttcg tctgccttct ttacgttcca taggccttaa 3360 gtgtttggag catttgtttt tctttcgcct tattggagat gttccaattg atacgttcct 3420 gatggagatg cttgaatcac cttctgattc ataatctagc ctagcccccc tctccctccc 3480 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata 3540 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg 3600 tc ttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt 3660 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag 3720 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc 3780 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga 3840 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg 3900 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat 3960 gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct 4020 ttgaaaaaca cgatctctag gcgccaccat gaagctactg tcttctatcg aacaagcatg 4080 cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg 4140 tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag 4200 ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat 4260 ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc 4320 attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt 4380 ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc 4440 atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgc cggaattccc 4500 ggggaiccgg cctgagtgcg tagtacccga gactcagtgc gccatgaagc ggaaagagaa 4560 gaaagcacag aaggagaagg acaaactgcc tgtcagcacg acgacggtgg acgaccacat 4620 gccgcccatt atgcagtgtg aacctccacc tcctgaagca gcaaggattc acgaagtggt 4680 cccaaggttt ctctccgaca agctgttggt gacaaaccgg cagaaaaaca -94 - tcccccagtt 4740 201207108

gacagccaac gccttctgat cgaagagtcg tatcgtggag tacgctgctt tgcggcctca caaggctggc tatggcgttg gccagggttg ccgcatctat gatcctctca ctccctcaag ggacatgtcg ctctagagcg gacaaaccac ttgctttatt attttatgtt acaaatgtgg aaacctctga gagcagacaa gaggtcgact gcaccaggtg tggatgtgac ttggctctag tgggaaagaa ccgccgccat gcatgccccc ccgtcctgcc agactgcagc actttgcttt acaagttgac ctcagcagct atgcggttta cttgctgtct cgttgagggt acatgggcat gggtggtgtt cagcagttcc gaagatttga gacactccct ttcgcgaagg aaggcttgct gacagtattc atggccgagg gacaacatcc gagcagccgc atcctgaacc atcctctctg ctcaagaaca cacacccaac gccgccaccg aactagaatg tgtaaccatt tcaggttcag tatggctgat cacatgcagc gcccgtcagg ctagtccccg cagaccctgc cgaggagctg cgatgaagat tatataaggt gagcaccaac atgggccggg cgcaaactct ctccgccgcc cctgagcccg ggctcttttg gttggatctg aaacataaat ttatttaggg cctgtgtatt aagcccgtct gtagatgatc ttatcgccag agaggattac tccgccagat gattgccagg caagtgaggt tgttcgcgaa tcatcgagga attacgcgct aactggtgga agctgagcgg agctacgcac gaaagctgcc cgccgcctat cggggagatc cagtgaaaaa ataagctgca ggggaggtgt tatgatccgg tcccggagac gcgcgtcagc cggtggcaga gagtgtggcg aggcccgatc acagattgag gggggtctta tcgtttgatg gtgcgtcaga actaccttga gcttcagccg cttgcaagca gcacaattgg cgccagcagg aaaaaaccag gttttgcgcg ttttccagga ctggggtgga cagtcgtagc gctcatctgg gcagacgtgg cacagagatg gttcgccaag aatgatgctc caaccaagcg tctactgcac gctcacggct agagatccag gtcggcgcgt gctcggcatg gcctttcctc cctcgagtcc cagacatgat aatgctttat ataaacaagt gggaggtttt ctgcctcgcg ggtcacagct gggtgttggc tctggaaggt gtaaacatat acttggtgct gtactgaaat tgtagttttg gaagcattgt atgtgatggg cctacgagac ctgcagccac gtgcagcttc attctttgac tttctgccct actctgtttg cgcggtaggc cgtggtaaag ggtagcacca aggagcgctg taccaggacg cagcaagcgg actatcctca atctcgcagc cgagtcgcgc tacactcgcg ttctgccggt gtcgtcatct cggtactacc tcgtccgtca caaaactcca gaggagatct cccacgaatc aagatacatt ttgtgaaatt taacaacaac ttaaagcaag cgtttcggtg tgtctgtaag gggtgtcggg gctgaggtac taggaaccag ggcctgcacc gtgtgggcgt tatctgtttt gagctcatat ctccagcatt cgtgtctgga cgcccgcggg ccgttcatcc ccgggaactt gaaggcttcc gatttggatc ccgggacca g gtgactctgg ctgcagagct ggcgtggtgc ggtacgagca acgatgaaaa cggtccaact ctgatcaaat gacgatacga acaactaccg gcatgtactc tttctgaccg tgaatacgct tatacggcaa acatgtgcat gggatgtggc tctaggcggc gatgagtttg tgtgatgcta aattgcattc taaaacctct atgacggtga cggatgccgg gcgcagccat gatgagaccc cctgtgatgc cgcgctgagt ggcttaaggg gcagcagccg ttgacaacgc gatggtcgcc acgccgttgg attgtgactg gcccgcgatg aatgtcgttt tcccctccca aagcaagtgt cggtctcggt atgttcagat tcatgctgcg ctaaaaatgt 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 -95 - 201207108 ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt 7020 taagctggga tgggtgcata cgtggggata tgagatgcat cttggactgt atttttaggt 7080 tggctatgtt cccagccata tccctccggg gattcatgtt Gtgcagaacc accagcacag 7140 tgtatccggt gcacttggga aatttgtcat gtagcttaga aggaaatgcg tggaagaact 7200 tggagacgcc cttgtgacct ccaagatttt ccatgcattc gtccataatg atggcaatgg 726 0 gcccacgggc ggcggcctgg gcgaagatat ttctgggatc actaacgtca tagttgtgtt 7320 ccaggatgag atcgtcatag gccattttta caaagcgcgg gcggagggtg ccagactgcg 7380 gtataatggt tccatccggc ccaggggcgt agttaccctc acagatttgc atttcccacg 7440 ctttgagttc agatgggggg atcatgtcta cctgcggggc gatgaagaaa acggtttccg 7500 gggtagggga gatcagctgg gaagaaagca ggttcctgag cagctgcgac ttaccgcagc 7560 cggtgggccc gtaaatcaca cctattaccg gctgcaactg gtagttaaga gagctgcagc 7620 tgccgtcatc cctgagcagg ggggccactt cgttaagcat gtccctgact cgcatgtttt 7680 ccctgaccaa atccgccaga aggcgctcgc cgcccagcga tagcagttct tgcaaggaag 7740 caaagttttt caacggtttg agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa 7800 gcagttccag gcggtcccac agctcggtca cctgctctac ggcatctcga tccagcatat 7860 ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc agtagtcggt gctcgtccag 7920 acgggccagg gtcatgtctt tccacgggcg cagggtcctc gtcagcgtag tctgggtcac 7980 ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct 8040 ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgacca tggt 8100 gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc agcttgccct tggaggaggc 8160 gccgcacgag gggcagtgca gacttttgag ggcgtagagc ttgggcgcga gaaataccga 8220 ttccggggag taggcatccg cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca 8280 ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc ccatgctttt 8340 cttacctctg gtttccatga gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc 8400 cccgtataca gacttgagag gcctgtcctc gaccgatgcc cttgagagcc ttcaacccag 8460 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct tgatgcgttt 8520 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc 8580 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg 8640 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca 8700 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag 8760 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt 8820 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc 8880 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatag g ctccgccccc 8940 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 9000 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgt t ccgaccctgc 9060 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag Cgtggcgctt tctcatagct 9120 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 9180 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt -96 - gagtccaacc 9240 201207108 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 9300 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa agccagttac 9360 ggacagtatt tggtatctgc gctctgctga taatcagtga ggcacctatc tcagcgatct cttcggaaaa agagttggta 9420 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 9480 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 9540 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 9600 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 9660 agtaaacttg gtctgacagt taccaatgct 9720 gtctatttcg Ttcatccata gttgcctgac tccc cgtcgt gtagataact acgatacggg 9780 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 9840 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 9900 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 9960 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt 10020 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 10080 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 10140 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 10200 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 10260 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata 10320 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 10380 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 10440 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 10500 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 10560 attgaagcat ttatcagggt tattgtctca Tgagcggata catatttgaa tgtatttaga 10620 aaaa 10624

C &lt;210&gt; 50 &lt;211&gt; 10790 &lt;212&gt; DNA &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; PolyDNA.from.V^ &lt;400&gt; 50 taaacaaata ggggttccgc cattattatc atgacattaa attaattaag ctagcatcat tgagggggtg gagtttgtga tgtggcggaa gtgtgatgtt aagtgacgtt tttggtgtgc ggcggatgtt gtagtaaatt actgaataag aggaagtgaa ^ -43321 5u2 gcacatttcc cctataaaaa caataatata cgtggcgcgg gcaagtgtgg gccggtgtac tgggcgtaac atctgaataa ccgaaaagtg taggcgtatc ccttattttg ggcgtgggaa cggaacacat acaggaagtg cgagtaagat ttttgtgtta ccacctgacg acgaggccct gattgaagcc cggggcgggt gtaagcgacg acaattttcg ttggccattt ctcatagcgc tctaagaaac ttcgtcttca aatatgataa gacgtagtag gatgtggcaa cgcggtttta tcgcgggaaa gtaatatttg 60 120 180 240 300 360 420 480 -97 - 201207108

tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 780 tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtcgaa gaaggtgagt aatcttaaca 1020 tgctcttttt tttttttttt gctaatccct tttgtgtgct gatgttagga tgacatttac 1080 aacaaatgtt tgttcctgac aggaaaaacc ttgctgggta ccttcgttgc cggacacttc 1140 ttgtcctcta ctttggaaaa aaggaattga gagccgctag cgccaccatg agcactgaaa 1200 gcatgatccg ggacgtggag ctggccgagg aagccctccc caagaaaacc ggcggccccc 1260 aggggagcag aagatgtttg ttcctgagcc tgttttcctt cctgatcgtg gcaggcgcta 1320 ccaccctgtt ctgcctgctg cactttggag tgatcggccc ccagagggag gagttcccca 1380 gggacctctc tctaatcagc cctctggccc aggcaggatc cgtcagatca tcttctcgaa 1440 ccccgagtga caagcctgta gcccatgttg tagcaaaccc tcaagccgag ggccagctcc 1500 agtggctgaa ccgccgggcc aatgccctgc tcgccaacgg cgtcgagctg agagataacc 1560 agctggtggt gccatcagag ggcctgtacc tcatctactc ccaggtcctg ttcaagggcc 1620 aaggctgccc ctccacccat gtgctcctca cccacaccat cagccgcatc gccgtgagct 1680 accagaccaa ggtcaacctc ctctctgcca tcaagagccc ctgccagagg gagaccccag 1740 agggggccga ggccaagccc tggtatgagc ccatctacct cggcggggtg ttccagctgg 1800 agaagggtga ccgactcagc gctgagatca atagacccga ctatctcgac tttgccgaga 1860 gcggccaggt gtactttggg atcat tgccc tgtgaatcga ttcgtacgtc gacatcgaga 1920 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 1980 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 2040 atcatgtctg ggcgcgccgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc 2100 tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttc cg 2160 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc 2220 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag 2280 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt 2340 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt 2400 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga 2460 gtagcgggct gctgggctgg gtacgtgcgc tcggggttgg cgagtgtgtt ttgtgaagtt 2520 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact 2580 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga gctagcgccg 2640 ccaccatggg ccctaaaaag aagcgtaaag tcgccccccc gaccgatgtc agcctggggg 2700 acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacggg -98 - ctagacgat t 2760 201207108

tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact 2820 ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg 2880 cccttggaat tgacgagtac ggtggggaat tcgagatgcc tgtggacagg atcctggagg 2940 cagagcttgc tgtggaacag aagagtgacc agggcgttga gggtcctggg ggaaccgggg 3000 gtagcggcag cagcccaaat gaccctgtga ctaacatctg tcaggcagct gacaaacagc 3060 tattcacgct tgttgagtgg gcgaagagga tcccacactt ttcctccttg cctctggatg 3120 atcaggtcat attgctgcgg gcaggctgga atgaactcct cattgcctcc ttttcacacc 3180 gatccattga tgttcgagat ggcatcctcc ttgccacagg tcttcacgtg caccgcaact 3240 cagcccattc agcaggagta ggagccatct ttgatcgggt gctgacagag ctagtgtcca 3300 aaatgcgtga catgaggatg gacaagacag agcttggctg cctgagggca atcattctgt 3360 ttaatccaga ggtgaggggt ttgaaatccg cccaggaagt tgaacttcta cgtgaaaaag 3420 tatatgccgc tttggaagaa tatactagaa caacacatcc cgatgaacca ggaagatttg 3480 caaaactttt gcttcgtctg ccttctttac gttccatagg ccttaagtgt ttggagcatt 3540 tgtttttctt tcgccttatt ggagatgttc caattgatac gttcctgatg gagatgctt g 3600 aatcaccttc tgattcataa tctagcctag cccccctctc cctccccccc ccctaacgtt 3660 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc 3720 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc 3780 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 3840 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg 3900 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat 3960 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga 4020 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc 4080 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg 4140 ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat 4200 ctctaggcgc caccatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac 4260 ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact 4320 gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag 4380 aagtggaatc aaggctagaa agactggaac agctatttct actgattttt c ctcgagaag 4440 accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat 4500 tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga 4560 ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga 4620 gtagtaacaa aggtcaaaga cagttgactg tatcgccgga attcccgggg atccggcctg 4680 agtgcgtagt acccgagact cagtgcgcca tgaagcggaa agagaagaaa gcacagaagg 4740 agaaggacaa actgcctgtc agcacgacga cggtggacga ccacatgccg cccattatgc 4800 agtgtgaacc tccacctcct gaagcagcaa ggattcacga agtggtccca aggtttctct 4860 ccgacaagct gttggtgaca aaccggcaga aaaacatccc ccagttgaca gccaaccagc 4920 agttccttat cgccaggctc atctggtacc aggacgggta cgagcagcct tctgatgaag 4980 -99 - 201207108 atttgaagag gattacgcag acgtggcagc aagcggacga tgaaaacgaa gagtcggaca 5040 ctcccttccg ccagatcaca gagatgacta tcctcacggt ccaacttatc gtggagttcg 5100 cgaagggatt gccagggttc gccaagatct cgcagcctga tcaaattacg ctgcttaagg 5160 cttgctcaag tgaggtaatg atgctccgag tcgcgcgacg atacgatgcg gcctcagaca 5220 gtattctgtt cgcgaacaac caagcgtacactcgcgacaa ctaccgcaag gctggcatgg 5280 ccgaggtcat cgaggatcta ctgcacttct gccggtgcat gtactctatg gcgttggaca 5340 acatccatta cgcgctgctc acggctgtcg tcatcttttc tgaccggcca gggttggagc 5400 agccgcaact ggtggaagag atccagcggt actacctgaa tacgctccgc atctatatcc 5460 tgaaccagct gagcgggtcg gcgcgttcgt ccgtcatata cggcaagatc ctctcaatcc 5520 tctctgagct acgcacgctc ggcatgcaaa actccaacat gtgcatctcc ctcaagctca 5580 agaacagaaa gctgccgcct ttcctcgagg agatctggga tgtggcggac atgtcgcaca 5640 cccaaccgcc gcctatcctc gagtccccca cgaatctcta ggcggcctct agagcggccg 5700 ccaccgcggg gagatccaga catgataaga tacattgatg agtttggaca aaccacaact 5760 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta 5820 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag 5880 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg 5940 gctgattatg atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 6000 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 6060 gtcagggcgc gtcagcgggt gtt ggcgggt gtcggggcgc agccatgagg tcgactctag 6120 tccccgcggt ggcagatctg gaaggtgctg aggtacgatg agacccgcac caggtgcaga 6180 ccctgcgagt gtggcggtaa acatattagg aaccagcctg tgatgctgga tgtgaccgag 6240 gagctgaggc ccgatcactt ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat 6300 gaagatacag attgaggtac tgaaatgtgt gggcgtggct taagggtggg aaagaatata 6360 taaggtgggg gtcttatgta gttttgtatc tgttttgcag cagccgccgc cgccatgagc 6420 accaactcgt ttgatggaag cattgtgagc tcatatttga caacgcgcat gcccccatgg 6480 gccggggtgc gtcagaatgt gatgggctcc agcattgatg gtcgccccgt cctgcccgca 6540 aactctacta ccttgaccta cgagaccgtg tctggaacgc cgttggagac tgcagcctcc 6600 gccgccgctt cagccgctgc agccaccgcc cgcgggattg tgactgactt tgctttcctg 6660 agcccgcttg caagcagtgc agcttcccgt tcatccgccc gcgatgacaa gttgacggct 6720 cttttggcac aattggattc tttgacccgg gaacttaatg tcgtttctca gcagctgttg 6780 gatctgcgcc agcaggtttc tgccctgaag gcttcctccc ctcccaatgc ggtttaaaac 6840 ataaataaaa aaccagactc tgtttggatt tggatcaagc aagtgtcttg ctgtctttat 6900 ttaggggttt tgcgcg cgcg gtaggcccgg gaccagcggt ctcggtcgtt gagggtcctg 6960 tgtatttttt ccaggacgtg gtaaaggtga ctctggatgt tcagatacat gggcataagc 7020 ccgtctctgg ggtggaggta gcaccactgc agagcttcat gctgcggggt ggtgttgtag 7080 atgatccagt cgtagcagga gcgctgggcg tggtgcctaa aaatgtcttt cagtagcaag 7140 ctgattgcca ggggcaggcc cttggtgtaa gtgtttacaa agcggttaag ctgggatggg 7200 tgcatacgtg gggatatgag atgcatcttg gactgtattt ttaggttggc -100 tatgttccca 7260 201207108

gccatatccc tccggggatt catgttgtgc agaaccacca gcacagtgta tccggtgcac 7320 ttgggaaatt tgtcatgtag cttagaagga aatgcgtgga agaacttgga gacgcccttg 7380 tgacctccaa gattttccat gcattcgtcc ataatgatgg caatgggccc acgggcggcg 7440 gcctgggcga agatatttct gggatcacta acgtcatagt tgtgttccag gatgagatcg 7500 tcataggcca tttttacaaa gcgcgggcgg agggtgccag actgcggtat aatggttcca 7560 tccggcccag gggcgtagtt accctcacag atttgcattt cccacgcttt gagttcagat 7620 ggggggatca tgtctacctg cggggcgatg aagaaaacgg tttccggggt aggggagatc 7680 agctgggaag aaagcaggtt cctgagcagc tgcgacttac cgcagccggt gggcccgtaa 7740 atcacaccta ttaccggctg caactggtag ttaagagagc tgcagctgcc gtcatccctg 7800 agcagggggg ccacttcgtt aagcatgtcc ctgactcgca tgttttccct gaccaaatcc 7860 gccagaaggc gctcgccgcc cagcgatagc agttcttgca aggaagcaaa gtttttcaac 7920 ggtttgagac cgtccgccgt aggcatgctt ttgagcgttt gaccaagcag ttccaggcgg 7980 tcccacagct cggtcacctg ctctacggca tctcgatcca gcatatctcc tcgtttcgcg 8040 ggttggggcg gctttcgctg tacggcagta gtcggtgctc gtccagacgg gccagggtc a 8100 tgtctttcca cgggcgcagg gtcctcgtca gcgtagtctg ggtcacggtg aaggggtgcg 8160 ctccgggctg cgcgctggcc agggtgcgct tgaggctggt cctgctggtg ctgaagcgct 8220 gccggtcttc gccctgcgcg tcggccaggt agcatttgac catggtgtca tagtccagcc 8280 cctccgcggc gtggcccttg gcgcgcagct tgcccttgga ggaggcgccg cacgaggggc 8340 agtgcagact tttgagggcg tagagcttgg gcgcgagaaa taccgattcc ggggagtagg 8400 catccgcgcc gcaggccccg cagacggtct cgcattccac gagccaggtg agctctggcc 8460 gttcggggtc aaaaaccagg tttcccccat gctttttgat gcgtttctta cctctggttt 8520 ccatgagccg gtgtccacgc tcggtgacga aaaggctgtc cgtgtccccg tatacagact 8580 tgagaggcct gtcctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg 8640 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc 8700 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc 8760 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc 8820 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg 8880 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg g atggccttc 8940 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg 9000 tccaggcagg tagatgacga ccatcaggga cagcttcaag gccagcaaaa ggccaggaac 9060 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 9120 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 9180 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 9240 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 9300 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 9360 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 9420 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 9480 101 201207108 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacc tagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcg Acacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa &lt;210&gt; 51 &lt;211&gt; 10790 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; PolyDNA_from_WN-43322 5U2 &lt;400> 51 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcgc Cgtttggccg cctcgagtct agagatccgg tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa -102 - 9540 9600 96 60 9720 9780 9840 9900 9960 10020 10080 10140 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10790 60 120 180 240 300 360 420 480 540 600 660 201207108

C

acaaaacaaa ctatcgataa tactgtcctc tgtcctccga ttccttcgaa ctggtgtgtg tgctcttttt aacaaatgtt ttgtcctcta gcatgatccg agggaagcag ccacactgtt gagacctgtc cccctagcga agtggctgaa agctcgtggt agggatgccc accaaaccaa agggagccga agaaaggcga gcggacaggt acttgtttat ataaagcatt atcatgtctg tcctcacggc cccggacgct agcagaagga agagcggaac ggggcggtga cgcagccggg gtagcgggct ttttaggcac agtaaattgt ccaccatggg acgagctcca tcgatctgga ccgcccccta ctagcaaaat tgcaggtcgg cgagcggagt gcggagactc ggaagagggg agctcatctt tttttttttt tgttcctgac ctttggaaaa ggacgtggag gagatgtctg ttgcctcctg cctgat tagc taagcctgtg tagaagggcc cccctccgag tagcacacac agtgaatctg agccaaaccc tagactcagc ctactttggc tgcagcttat tttttcactg ggcgcgccgg gagcgctgcc caggacagcg cattttagga aggcgaggaa acgccgatga atttgggtcg gctgggctgg cttttgaaat ccgctaaatt ccctaaaaag cttagacggc catgttgggg cggcgctctg aggctgtccc agtactgtcc actgtcctcc ttcgaaggaa cggggtcgaa cctgtagatc gctaatccct aggaaaaacc aaggaattga ctggccgagg tttctgtccc catttcggag cctctggctc gctcacgtcg aatgccctgc ggactgtatc gtcctgctg a ctgtccgcta tggtacgaac gctgagatta attatcgctc aatggttaca cattctagtt cctccgcgcc acgtcagacg gcccgctgct cgggacttgg aagtagtccc ttatataagg cggttcttgt gtacgtgcgc gtaatcattt ctggccgttt aagcgtaaag gaggacgtgg gacggggatt gatatggccg cagtgcaagt tccgagcgga gagcggagta gaggggcggg gacctagagg acgcgtcgaa tttgtgtgct ttgctgggta gagccgctag aagccctgcc tgtttagctt tgattggccc aggctggatc tcgctaaccc tcgccaacgg tgatttactc cccacaccat tcaaaagccc ccatttacct acaggcccga tgtaaatcga aataaagcaa gtggtttgtc gggttttggc aagggcgcag cataagactc gtgactctag ttctcggcga acgcgccggg ttgtggatcg tcggggttgg gggtcaatat ttggcttttt tcgccccccc cgatggcgca ccccgggtcc acttcgagtt gcaggtgcca gtactgtcct ctgtcctccg gtcgatcgac gtatataatg gaaggtgagt gatgttagga ccttcgttgc cgccaccatg taagaaaacc tctgattgtg tcagagggag cgtgagaagc tcaggccgag cgtcgagctg ccaggtcctg tagcaggatc ttgccaaaga cggcggagtg ttacctcgac ttcgtacgtc tagcatcaca caaactcatc gcctcccgcg cgagcgtcct ggccttagaa ggcactggtt ttctgcggag tgtggcacag ctgtgatcgt cgagtgtgtt gtaattttca tgttagacga gaccgatg tc tgccgacgcg gggatttacc tgagcagatg gaacatttct ccgagcggag agcggagtac cccgcccctc ggtgccttag aatcttaaca tgacatttac cggacacttc tccaccgaaa ggaggccctc gctggcgcta gagttcccta agcagcagga ggccagctcc agagacaatc tttaagggac gctgtgtcct gaaacccctg tttcagctgg tttgccgaaa gacatcgaga aatttcacaa aatgtatctt ggcgcccccc gatccttccg ccccagtatc ttctttccag ggatctccgt ctagttccgt cacttggtga ttgtgaagtt gtgttagact gctagcgccg agcctggggg ctagacgatt ccccacgact tttaccgatg 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 -103 - 201207108 cccttggaat tgacgagtac ggtggggaat tcgagatgcc tgtggacagg atcctggagg 2940 cagagcttgc tgtggaacag aagagtgacc agggcgttga gggtcctggg ggaaccgggg 3000 gtagcggcag cagcccaaat gaccctgtga ctaacatctg Tcaggcagct gacaaacagc 3060 tattcacgct tgttgagtgg gcgaagagga tcccacactt ttcctccttg cctctggatg 3120 atcaggtcat attgctgcgg gcaggctgga atgaactcct cattgcctcc ttttcacacc 3180 gatccattga tgttcgagat ggcatcctcc ttgccacagg tcttcacgtg caccgcaact 3240 cagcccattc agcaggagta ggagccatct ttgatcgggt gctgacagag ctagtgtcca 3300 aaatgcgtga catgaggatg gacaagacag agcttggctg cctgagggca atcattctgt 3360 ttaatccaga ggtgaggggt ttgaaatccg cccaggaagt tgaacttcta cgtgaaaaag 3420 tatatgccgc tttggaagaa tatactagaa caacacatcc cgatgaacca ggaagatttg 3480 caaaactttt gcttcgtctg ccttctttac gttccatagg ccttaagtgt ttggagcatt 3540 tgtttttctt tcgccttatt ggagatgttc caattgatac gttcctgatg gagatgcttg 3600 aatcaccttc tgattcataa tctagcctag cccccctctc cctccccccc ccctaacgtt 3660 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc 3720 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc 3780 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 3840 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg 3900 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat 3960 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaag a 4020 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc 4080 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg 4140 ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat 4200 ctctaggcgc caccatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac 4260 t taaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact 4320 gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag 4380 aagtggaatc aaggctagaa agactggaac agctatttct actgattttt cctcgagaag 4440 accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat 4500 tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga 4560 ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga 4620 gtagtaacaa aggtcaaaga cagttgactg tatcgccgga attcccgggg atccggcctg 4680 agtgcgtagt acccgagact cagtgcgcca tgaagcggaa agagaagaaa gcacagaagg 4740 agaaggacaa actgcctgtc agcacgacga cggtggacga ccacatgccg cccattatgc 4800 agtgtgaacc tccacctcct gaagcagcaa ggattcacga agtggtccca aggtttctct 4860 ccgacaagct gttggtgaca aaccggcaga aaaacatccc ccagttgaca gccaaccagc 4920 agttccttat cgccaggctc atctggtacc aggacgggta cgagcagcct tctgaigaag 4980 atttgaagag gattacgcag acgtggcagc aagcggacga tgaaaacgaa gagtcggaca 5040 ctcccttccg ccagatcaca gagatgacta tcctcacggt ccaacttatc gtggagttcg 5100 cgaagggatt gccagggttc gccaagatct cgcagcctga tcaaat tacg -104 - ctgcttaagg 5160 201207108 ()

cttgctcaag gtattctgtt ccgaggtcat acatccatta agccgcaact tgaaccagct tctctgagct agaacagaaa cccaaccgcc ccaccgcggg agaatgcagt accattataa gttcaggggg gctgattatg tgcagctccc gtcagggcgc tccccgcggt ccctgcgagt gagctgaggc gaagatacag taaggtgggg accaactcgt gccggggtgc aactctacta gccgccgctt agcccgcttg cttttggcac gatctgcgcc ataaataaaa ttaggggttt tgtatttttt ccgtctctgg atgatccagt ctgattgcca tgcatacgtg gccatatccc ttgggaaatt tgaggtaatg cgcgaacaac cgaggatcta cgcgctgctc ggtggaagag gagcgggtcg acgcacgctc gctgccgcct gcctatcctc gagatccaga gaaaaaaatg gctgcaataa aggtgtggga atccggctgc ggagacggtc gtcagcgggt ggcagatctg gtggcggtaa ccgatcactt attgaggtac gtcttatgta ttgatggaag gtcagaatgt ccttgaccta cagccgctgc caagcagtgc aattggattc agcaggtttc aaccagactc tgcgcgcgcg ccaggacgtg ggtggaggta cgtagcagga ggggcaggcc gggatatgag tccggggatt tgtcatgtag atgctccgag caagcgtaca ctgcacttct acggctgtcg atccagcggt gcgcgttcgt ggcatgcaaa ttcctcgagg gagtccccca catgataaga ctttatttgt acaagttaac ggttttttaa ctcgcgcgtt acagcttgtc gttggcgggt gaaggtgctg acatattagg ggtgctggcc tgaaatgtgt gttttgtatc cattgtgagc gatgggctcc cgagaccgtg agccaccgcc agcttcccgt tttgacccgg tgccctgaag tgtttggatt gtaggcccgg gtaaaggtga gcaccactgc gcgctgggcg cttggtgtaa atgcatcttg catgttgtgc cttagaagga tcgcgcgacg ctcgcgacaa gccggtgcat tcatcttttc actacctgaa ccgtcatata actccaacat agatctggga cgaatctcta tacattgatg gaaatttgtg aacaacaatt agcaagtaaa tcggtgatga tgtaagcgga gtcggggcgc aggtacgatg aaccagcctg tgcacccgcg gggcgtggct tgttttgcag tcatatttga agcattgatg tctggaacgc cgcgggattg tcatccgccc gaacttaatg gcttcctccc tggatcaagc gaccagcggt ctctggatgt agagcttcat tggtgcctaa gtgtttacaa gactgtattt agaaccacca aatgcgtgga atacgatgcg ctaccgcaag gtactctatg tgaccggcca tacgctccgc cggcaagatc gtgcatctcc tgtggcggac ggcggcctct agtttggaca atgctattgc gcattcattt acctctacaa cggtgaaaac tgccgggagc agccatgagg agacccgcac tgatgctgga ctgagtttgg taagggtggg cagccgccgc caacgcgcat gtcgccccgt cgttggagac tgactgactt gcgatgacaa tcgtttctca ctcccaatgc aagtgtcttg ctcggtcgtt tcagatacat gctgcggggt aaatgtcttt agcggttaa g ttaggttggc gcacagtgta agaacttgga gcctcagaca gctggcatgg gcgttggaca gggttggagc atctatatcc ctctcaatcc ctcaagctca atgtcgcaca agagcggccg aaccacaact tttatttgta tatgtttcag atgtggtatg ctctgacaca agacaagccc tcgactctag caggtgcaga tgtgaccgag ctctagcgat aaagaatata cgccatgagc gcccccatgg cctgcccgca tgcagcctcc tgctttcctg gttgacggct gcagctgttg ggtttaaaac ctgtctttat gagggtcctg gggcataagc ggtgttgtag cagtagcaag ctgggatggg tatgttccca tccggtgcac gacgccctig 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 -105 - 201207108 tgacctccaa gattttccat gcattcgtcc ataatgatgg caatgggccc acgggcggcg 7440 gcctgggcga agatatttct gggatcacta acgtcatagt tgtgttccag gatgagatcg 7500 tcataggcca tttttacaaa gcgcgggcgg agggtgccag Actgcggtat aatggttcca 7560 tccggcccag gggcgtagtt accctcacag atttgcattt cccacgcttt gagttcagat 7620 ggggggatca tgtctacctg cggggcgatg aagaaaacgg tttccggggt aggggagatc 76 80 agctgggaag aaagcaggtt cctgagcagc tgcgacttac cgcagccggt gggcccgtaa 7740 atcacaccta ttaccggctg caactggtag ttaagagagc tgcagctgcc gtcatccctg 7800 agcagggggg ccacttcgtt aagcatgtcc ctgactcgca tgttttccct gaccaaatcc 7860 gccagaaggc gctcgccgcc cagcgatagc agttcttgca aggaagcaaa gtttttcaac 7920 ggtttgagac cgtccgccgt aggcatgctt ttgagcgttt gaccaagcag ttccaggcgg 7980 tcccacagct cggtcacctg ctctacggca tctcgatcca gcatatctcc tcgtttcgcg 8040 ggttggggcg gctttcgctg tacggcagta gtcggtgctc gtccagacgg gccagggtca 8100 tgtctttcca cgggcgcagg gtcctcgtca gcgtagtctg ggtcacggtg aaggggtgcg S160 ctccgggctg cgcgctggcc agggtgcgct tgaggctggt cctgctggtg ctgaagcgct 8220 gccggicttc gccctgcgcg tcggccaggt agcatttgac catggtgtca tagtccagcc S280 cctccgcggc gtggcccttg gcgc £ cagct tgcccttgga ggaggcgccg cacgaggggc 8340 agtgcagact tttgagggcg tagagcttgg gcgcgagaaa taccgattcc ggggagtagg 8400 catccgcgcc gcaggccccg cagacggtct cgcattccac gagccaggtg agctctggcc 8460 gttcggggtc aaaaaccagg tttcccccat gctttttgat gcgtttctta cctc tggttt 8520 ccatgagccg gtgtccacgc tcggtgacga aaaggctgtc cgtgtccccg tatacagact 8580 tgagaggcct gtcctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg 8640 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc 8700 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc 8760 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc 8820 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg 8880 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc 8940 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg 9000 tccaggcagg tagatgacga ccatcaggga cagcttcaag gccagcaaaa ggccaggaac 9060 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 9120 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 9180 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 9240 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 9300 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacga taccggatac acc ccccgttcag 9360 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 9420 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 9480 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 9540 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 9600 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat -106 - tacgcgcaga 9660 201207108

aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 9720 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 9780 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 9840 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 9900 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 9960 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 10020 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 10080 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 10140 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct 10200 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 10260 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 10320 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 10380 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 10440 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa 105 00 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 10560 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 10620 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 10680 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 10740 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 10790 &lt; 210> 52 &lt; 211 &gt; 10575 &lt; 212 &gt; DNA cggggcgggt gacgtagtag 52 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 60 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaattaag ctagcatcat caataatata ccttatttig gattgaagcc aatatgataa 180 tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa; &lt; 213 &gt; artificial organisms &lt; 220 &gt; &lt; 223 &gt; PolyDNA_from_WN-43323 5U2 &lt; 400 & gt 240 tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa 300 aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta 360 ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa 420 actgaataag aggaagtg aa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg 480 tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 780 • 107 · 201207108 tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtcgaa gaaggtgagt aatcttaaca 1020 tgctcttttt tttttttttt gctaatccct tttgtgtgct gatgttagga tgacatttac 1080 aacaaatgtt tgttcctgac aggaaaaacc ttgctgggta ccttcgttgc cggacacttc 1140 ttgtcctcta ctttggaaaa aaggaattga gagccgctag cgccaccatg tatagaatgc 1200 agctcctgtc ctgcattgcc ctgagcctcg ccctcgtgac aaactccgcc cctaccagcg 1260 gatcc gtgag aagcagcagc aggaccccta gcgataagcc tgtggctcac gtcgtcgcta 1320 accctcaggc cgagggccag ctccagtggc tgaatagaag ggccaatgcc ctgctcgcca 1380 acggcgtcga gctgagagac aatcagctcg tggtcccctc cgagggactg tatctgattt 1440 actcccaggt cctgtttaag ggacagggat gccctagcac acacgtcctg ctgacccaca 1500 ccattagcag gatcgctgtg tcctaccaaa ccaaagtgaa tctgctgtcc gctatcaaaa 1560 gcccttgcca aagagaaacc cctgagggag ccgaagccaa accctggtac gaacccattt 1620 acctcggcgg agtgtttcag ctggagaaag gcgatagact cagcgctgag attaacaggc 1680 ccgattacct cgactttgcc gaaagcggac aggtctactt tggcattatc gctctgtaaa 1740 tcgattcgta cgtcgacatc gagaacttgt ttattgcagc ttataatggt tacaaataaa 1800 gcaatagcat cacaaat t tc acaaataaag catttttttc actgcattct agttgtggtt 1860 tgtccaaact catcaatgta tcttatcatg tctgggcgcg ccggcctccg cgccgggttt 1920 tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc 1980 gcagcgagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag 2040 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact 2 100 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg 2160 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc 2220 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg 2280 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctgggtacgt gcgctcgggg 2340 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca 2400 atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct 2460 tttttgttag acgagctagc gccgccacca tgggccctaa aaagaagcgt aaagtcgccc 2520 ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac gtggcgatgg 2580 cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg gat tccccgg 2640 gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg 2700 agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtggg gaattcgaga 2760 tgcctgtgga caggatcctg gaggcagagc ttgctgtgga acagaagagt gaccagggcg 2820 ttgagggtcc tgggggaacc gggggtagcg gcagcagccc aaatgaccct gtgactaaca 2880 ictgtcaggc agctgacaaa cagctattca cgctigttga gtgggcgaag agg Atcccac 2940 acttttcctc cttgcctctg gatgatcagg tcatattgct gcgggcaggc tggaatgaac 3000 tcctcattgc ctccttttca caccgatcca ttgatgttcg agaiggcatc ctccttgcca 3060 -108 -

201207108 caggtcttca cgtgcaccgc aactcagccc attcagcagg agtaggagcc atctttgatc gggtgctgac agagctagtg tccaaaatgc gtgacatgag gatggacaag acagagcttg gctgcctgag ggcaatcatt ctgtttaatc cagaggtgag gggtttgaaa tccgcccagg aagttgaact tctacgtgaa aaagtatatg ccgctttgga agaatatact agaacaacac atcccgatga accaggaaga tttgcaaaac ttttgcttcg tctgccttct ttacgttcca taggccttaa gtgtttggag catttgtttt tctttcgcct tattggagat gttccaattg atacgttcct gatggagatg cttgaatcac cttctgattc ataatctagc ctagcccccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatctctag gcgccaccat gaagctactg tcttctatcg aacaagcatg cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgc cggaattccc ggggatccgg cctgagtgcg tagtacccga gactcagtgc gccatgaagc ggaaagagaa gaaagcacag aaggagaagg acaaactgcc tgtcagcacg acgacggtgg acgaccacat gccgcccatt atgcagtgtg aacctccacc tcctgaagca gcaaggattc acgaagtggt cccaaggttt ctctccgaca agctgttggt gacaaaccgg cagaaaaaca tcccccagtt gacagccaac cagcagttcc ttatcgccag gctcatctgg taccaggacg ggtacgagca gccttctgat gaagatttga agaggattac gcagacgtgg cagcaagcgg acgatgaaaa cgaagagtcg gacactccct tccgccagat cacagagatg actatcctca cggtccaact tatcgtggag ttcgcgaagg gattgccagg gttcgccaag atctcgcagc ctgatcaaat tacgctgctt aaggcttgct caagtgaggt aatgatgctc cgagtcgcgc gacgatacga tgcggcctca gacagtattc tgttcgcgaa caaccaagcg tacactcgcg acaactaccg caaggctggc atggccgagg tcatcgagga tctactgcac ttctgccggt gcatgtactc tatggcgttg gacaacatcc attacgcgct gctcacggct gtcgtcatct tttctgaccg gccagggttg gagcagccgc aactggtgga agagatccag cggtactacc 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 109 - 201207108 tgaatacgct ccgcatctat atcctgaacc agctgagcgg gtcggcgcgt tcgtccgtca 5340 tatacggcaa gatcctctca atcctctctg agctacgcac gctcggcatg caaaactcca 5400 acatgtgcat ctccctcaag ctcaagaaca gaaagctgcc gcctttcctc Gaggagatct 5460 gggatgtggc ggacatgtcg cacacccaac cgccgcctat cctcgagtcc cccacgaatc 5520 tctaggcggc ctctagagcg gccgccaccg cggggagatc cagacatgat aaga tacatt 5580 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt 5640 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac 5700 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag 5760 taaaacctct acaaatgtgg tatggctgat tatgatccgg ctgcctcgcg cgtttcggtg 5820 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 5880 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 5940 gcgcagccat gaggtcgact ctagtccccg cggtggcaga tctggaaggt gctgaggtac 6000 gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat taggaaccag 6060 cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct ggcctgcacc 6120 cgcgctgagt ttggctctag cgatgaagat acagattgag gtactgaaat gtgtgggcgt 6180 ggcttaaggg tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt 6240 gcagcagccg ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat 6300 ttgacaacgc gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt 6360 gatggtcgcc ccgtcctgcc cgcaaactct actaccttga cctacga gac cgtgtctgga 6420 acgccgttgg agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg 6480 attgtgactg actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc 6540 gcccgcgatg acaagt tgac ggctcttttg gcacaattgg attctttgac ccgggaactt 6600 aatgtcgttt ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc 6660 tcccctccca atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc 6720 aagcaagtgt cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag 6780 cggtctcggt cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg 6840 atgttcagat acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct 6900 tcatgctgcg gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc 6960 ctaaaaatgt ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt 7020 acaaagcggt taagctggga tgggtgcata cgtggggata tgagatgcat cttggactgt 7080 atttttaggt tggctatgtt cccagccata tccctccggg gattcatgtt gtgcagaacc 7140 accagcacag tgtatccggt gcacttggga aatttgtcat gtagcttaga aggaaatgcg 7200 tggaagaact tggagacgcc cttgtgacct ccaagattt t ccatgcattc gtccataatg 7260 atggcaatgg gcccacgggc ggcggcctgg gcgaagatat ttctgggatc actaacgtca 7320 tagttgtgtt ccaggatgag atcgtcatag gccattttta caaagcgcgg gcggagggtg 7380 ccagactgcg gtataatggt tccatccggc ccaggggcgt agttaccctc acagatttgc 7440 atttcccacg ctttgagttc agatgggggg atcatgtcta cctgcggggc gatgaagaaa 7500 acggtttccg gggtagggga gatcagctgg gaagaaagca ggttcctgag • 110 - cagctgcgac 7560

201207108 ttaccgcagc cggtgggccc gtaaatcaca cctattaccg gctgcaactg gtagttaaga gagctgcagc tgccgtcatc cctgagcagg ggggccactt cgttaagcat gtccctgact cgcatgtttt ccctgaccaa atccgccaga aggcgctcgc cgcccagcga tagcagttct tgcaaggaag caaagttttt caacggtttg agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa gcagttccag gcggtcccac agctcggtca cctgctctac ggcatctcga tccagcatat ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc agtagtcggt gctcgtccag acgggccagg gtcatgtctt tccacgggcg cagggtcctc gtcagcgtag tctgggtcac ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc agcttgccct tggaggaggc gccgcacgag gggcagtgca gacttttgag ggcgtagagc ttgggcgcga gaaataccga ttccggggag taggcatccg cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt cttacctctg gtttccatga gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc cccgtataca gacttgagag gcctgtcctc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct itatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 9720 9780 111 - 201207108 acgatacggg agggcttacc tcaccggctc cagatttatc ggtcctgcaa ctttatccgc agtagttcgc cagttaatag tcacgctcgt cgtttggtat acatgatccc ccatgttgtg agaagtaagt tggccgcagt actgtcatgc catccgtaag tgagaatagt gtatgcggcg gcgccacata Gcagaacttt ctctcaagga tcttaccgct tgatcttcag catcttttac aatgccgcaa aaaagggaat tttcaatatt attga atctggcccc agtgctgcaa agcaataaac cagccagccg ctccatccag tctattaatt tttgcgcaac gttgttgcca ggcttcattc agctccggtt caaaaaagcg gttagctcct gttatcactc atggttatgg atgcttttct gtgactggtg accgagttgc tcttgcccgg aaaagtgctc atcattggaa gttgagatcc agttcgatgt tttcaccagc gtttctgggt aagggcgaca cggaaatgtt tgataccgcg agacccacgc gaagggccga gcgcagaagt gttgccggga agctagagta ttgctgcagg catcgtggtg cccaacgatc aaggcgagtt tcggtcctcc gatcgttgtc cagcactgca taattctctt agtactcaac caagtcattc cgicaacacg ggataatacc aacgttcttc ggggcgaaaa aacccactcg tgcacccaac gagcaaaaac aggaaggcaa gaatactcat actcttcctt 9840 9900 9960 10020 10080 10140 10200 10260 10320 10380 10440 10500 10560 10575 &lt;210&gt; 53 &lt;211&gt; 11276 &lt;212&gt; DNA &lt;213&gt;Artificial sequence&lt;220&gt;&lt;223&gt; PolyDNA-from-WN-43324 &lt;400&gt; 53 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgtggcggggg ggcgtgggaa cggggcggg t gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca catcccctga caagctgcca ggcaggttct cttcctctca catactgacc cacggctc Ca -112 - 60 320 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 201207108

C (ccctctctcc cctggaaagg acaccgctag cgccaccatg agcactgaaa gcatgatccg 1200 ggacgtggag ctggccgagg aagccctccc caagaaaacc ggcggccccc aggggagcag 1260 aagatgtttg ttcctgagcc tgttttcctt cctgatcgtg gcaggcgcta ccaccctgtt 1320 ctgcctgctg cactttggag tgatcggccc ccagagggag gagttcccca gggacctctc 1380 tctaatcagc cctctggccc aggcaggatc cgtcagatca tcttctcgaa ccccgagtga 1440 caagcctgta gcccatgttg tagcaaaccc tcaagccgag ggccagctcc agtggctgaa 1500 ccgccgggcc aatgccctgc tcgccaacgg cgtcgagctg agagataacc agctggtggt 1560 gccatcagag ggcctgtacc tcatctactc ccaggtcctg ttcaagggcc aaggctgccc 1620 ctccacccat gtgctcctca cccacaccat cagccgcatc gccgtgagct accagaccaa 1680 ggtcaacctc ctctctgcca tcaagagccc ctgccagagg gagaccccag agggggccga 1740 ggccaagccc tggtatgagc ccatctacct cggcggggtg ttccagctgg agaagggtga 1800 ccgactcagc gctgagatca atagacccga ctatctcgac tttgccgaga gcggccaggt 1860 gtactttggg atcattgccc tgtgaatcga ttcgtacgtc gacatcgagt gatgggtggc 1920 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagt gccca 1980 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata 2040 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta 2100 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg 2160 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat 2220 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca 2280 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc 2340 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa 2400 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac 2460 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt 2520 agatgcctgt tgaattggcg cgccggcctc cgcgccgggt tttggcgcct cccgcgggcg 2580 cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc 2640 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc 2700 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct 2760 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgatt ct gcggagggat 2820 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag 2880 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact 2940 tggtgagtag cgggctgctg ggctgggtac gtgcgctcgg ggttggcgag tgtgttttgt 3000 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt 3060 tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgagcta 3120 gcgccgccac catgggccct aaaaagaagc gtaaagtcgc ccccccgacc gatgtcagcc 3180 tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag 3240 acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga tttacccccc 3300 acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag cagatgttta 3360 .113 - 201207108 ccgatgccct tggaattgac gagtacggtg gggaattcga gatgcctgtg gacaggatcc 3420 tggaggcaga gcttgctgtg gaacagaaga gtgaccaggg cgttgagggt cctgggggaa 3480 ccgggggtag cggcagcagc ccaaatgacc ctgtgactaa catctgtcag gcagctgaca 3540 aacagctatt cacgcttgtt gagtgggcga agaggatccc acacttttcc tccttgcctc 3600 tggatgatca ggtcatattg Ctgcgg gcag gctggaatga actcctcatt gcctcctttt 3660 cacaccgatc cattgatgtt cgagatggca tcctccttgc cacaggtctt cacgtgcacc 3720 gcaactcagc ccattcagca ggagtaggag ccatctttga tcgggtgctg acagagctag 3780 tgtccaaaat gcgtgacatg aggatggaca agacagagct tggctgcctg agggcaatca 3840 ttctgtttaa tccagaggtg aggggtttga aatccgccca ggaagttgaa cttctacgtg 3900 aaaaagtata tgccgctttg gaagaatata ctagaacaac acatcccgat gaaccaggaa 3960 gatttgcaaa acttttgctt cgtctgcctt ctttacgttc cataggcctt aagtgtttgg 4020 agcatttgtt tttctttcgc cttattggag atgttccaat tgatacgttc ctgatggaga 4080 tgcttgaatc accttctgat tcataatcta gcctagcccc cctctccctc ccccccccct 4140 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt 4200 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg 4260 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc 4320 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccct t 4380 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta 4440 taagatacac ctgcaaag gc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg 4500 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag 4560 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag 4620 tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa 4680 cacgatctct aggcgccacc atgaagctac tgtcttctat cgaacaagca tgcgatattt 4740 gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga 4800 acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc 4860 tgacagaagt ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc 4920 gagaagacct tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa 4980 caggattatt tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag 5040 tggagactga tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg 5100 aagagagtag taacaaaggt caaagacagt tgactgtatc gccggaattc ccggggatcc 5160 ggcctgagtg cgtagtaccc gagactcagt gcgccatgaa gcggaaagag aagaaagcac 5220 agaaggagaa ggacaaactg cctgtcagca cgacgacggt ggacgaccac atgccgccca 5280 ttatgcagtg tgaacctcca cctcctgaag cagcaaggat tcacgaagtg gtcccaaggt 5340 ttctctccga caagctgttg gtgacaaacc ggcagaaaaa catcccccag ttgacagcca 5400 accagcagtt ccttatcgcc aggctcatct ggtaccagga cgggtacgag cagccttctg 5460 atgaagattt gaagaggatt acgcagacgt ggcagcaagc ggacgatgaa aacgaagagt 5520 cggacactcc cttccgccag atcacagaga tgactatcct cacggtccaa cttatcgtgg 5580 agttcgcgaa gggattgcca gggttcgcca agatctcgca gcctgatcaa -114 - attacgctgc 5640 201207108

ttaaggcttg cagacagtat gcatggccga tggacaacat tggagcagcc atatcctgaa caatcctctc agctcaagaa cgcacaccca cggccgccac acaactagaa tttgtaacca tttcaggttc ggtatggctg gacacatgca aagcccgtca ctctagtccc tgcagaccct accgaggagc agcgatgaag aatatataag atgagcacca ccatgggccg cccgcaaact gcctccgccg ttcctgagcc acggctcttt ctgttggatc taaaacataa ctttatttag gtcctgtgta ataagcccgt ttgtagatga agcaagctga gatgggtgca ttcccagcca gtgcacttgg ctcaagtgag tctgttcgcg ggtcatcgag ccattacgcg gcaactggtg ccagctgagc tgagctacgc cagaaagctg accgccgcct cgcggggaga tgcagtgaaa ttataagctg agggggaggt attatgatcc gctcccggag gggcgcgtca cgcggtggca gcgagtgtgg tgaggcccga atacagattg gtgggggtct actcgtttga gggtgcgtca ctactacctt ccgcttcagc cgcttgcaag tggcacaatt tgcgccagca ataaaaaacc gggttttgcg ttttttccag ctctggggtg tccagtcgta ttgccagggg tacgtgggga acgctcggca tatccctccg gaaatttgtc gtaatgatgc aacaaccaag gatctactgc ctgctcacgg gaagagatcc gggtcggcgc gtgggaggtt ggctgcctcg acggtcacag ccgcctttcc atcctcgagt tccagacatg aaaatgcttt caataaacaa gcgggtgttg gatctggaag cggtaaacat tcacttggtg aggtactgaa tatgtagttt tggaagcatt gaatgtgatg gacctacgag cgctgcagcc cagtgcagct ggattctttg ggtttctgcc agactctgtt cgcgcggtag gacgtggtaa gaggtagcac gcaggagcgc caggcccttg tatgagatgc gggattcatg atgtagctta tccgagtcgc cgtacactcg acttctgccg ctgtcgtcat agcggtacta gttcgtccgt tgcaaaactc tcgaggagat cccccacgaa ataagataca at ttgtgaaa gttaacaaca ttttaaagca cgcgtttcgg cttgtctgta gcgggtgtcg gtgctgaggt attaggaacc ctggcctgca atgtgtgggc tgtatctgtt gtgagctcat ggctccagca accgtgtctg accgcccgcg tcccgttcat acccgggaac ctgaaggctt tggatttgga gcccgggacc aggtgactct cactgcagag tgggcgtggt gtgtaagtgt atcttggact ttgtgcagaa gaaggaaatg gcgacgatac cgacaactac gtgcatgtac cttttctgac cctgaatacg catatacggc caacatgtgc ctgggatgtg tctctaggcg ttgatgagtt tttgtgatgc acaattgcat agtaaaacct tgatgacggt agcggatgcc gggcgcagcc acgatgagac agcctgtgat cccgcgctga gtggcttaag ttgcagcagc atttgacaac ttgatggtcg gaacgccgtt ggattgtgac ccgcccgcga ttaatgtcgt cctcccctcc tcaagcaagt agcggtctcg ggatgttcag cttcatgctg gcctaaaaat ttacaaag cg gtatttttag ccaccagcac cgtggaagaa gatgcggcct cgcaaggctg tctatggcgt cggccagggt ctccgcatct aagatcctct atctccctca gcggacatgt gcctctagag tggacaaacc tattgcttta tcattttatg ctacaaatgt gaaaacctct gggagcagac atgaggtcga ccgcaccagg gctggatgtg gtttggctct ggtgggaaag cgccgccgcc gcgcatgccc ccccgtcctg ggagactgca tgactttgct tgacaagttg ttctcagcag caatgcggtt gtcttgctgt gtcgttgagg atacatgggc cggggtggtg gtctttcagt gttaagctgg gttggctatg agtgtatccg cttggagacg 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 • 115 - 201207108 cccttgtgac ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg 7920 gcggcggcct gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg 7980 agatcgtcat aggccatttt tacaaagcgc gggcggaggg Tgccagactg cggtataatg 8040 gttccatccg gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt 8100 tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtagg g 8160 gagatcagct gggaagaaag caggttcctg agcagctgcg acttaccgca gccggtgggc 8220 ccgtaaatca cacctattac cggctgcaac tggtagttaa gagagctgca gctgccgtca 8280 tccctgagca ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc 8340 aaatccgcca gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt 8400 ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc 8460 aggcggtccc acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt 8520 ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca 8580 gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg 8640 ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga 8700 agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt 8760 ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcacg 8820 aggggcagtg cagact 11 tg agggcgtaga gcttgggcgc gagaaatacc gattccgggg 8880 agtaggcatc cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct 8940 ctggccgttc ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc 9000 tggtttccat gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata 9060 cagacttgag aggcctgtcc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc 9120 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg 9180 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc 9240 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct 9300 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc 9360 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg 9420 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc 9480 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggcca gcaaaaggcc 9540 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 9600 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 9660 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 9720 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 9780 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gc tgtgtgca cgaacccccc 9840 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 9900 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 9960 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 10020 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 10080 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 10140

201207108 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaa &lt; 210 &gt; 54 &lt; 211> 11276 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; PolyDNA_from_WN-43325 &lt; 400 &gt; 54 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaac Cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcc cgtttggccg cctcgagtct agagatccgg tgagtattag gc gcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 11160 11220 11276 60 120 180 240 300 360 420 480 540 600 660 720 780 117 - 201207108 tactgicctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg 1020 accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca 1080 catcccctga caagctgcca ggcaggttct cttcctctca catactgacc cacggctcca 1140 Ccctctctcc cctggaaagg acaccgctag cgccaccatg tccaccgaaa gcatgatccg 1200 ggacgtggag ctggccgagg aagccctgcc taagaaaacc ggaggccctc agggaagcag 1 260 gagatgtctg tttctgtccc tgtttagctt tctgattgtg gctggcgcta ccacactgtt 1320 ttgcctcctg catttcggag tgattggccc tcagagggag gagttcccta gagacctgtc 1380 cctgattagc cctctggctc aggctggatc cgtgagaagc agcagcagga cccctagcga 1440 taagcctgtg gctcacgtcg tcgctaaccc tcaggccgag ggccagctcc agtggctgaa 1500 tagaagggcc aatgccctgc tcgccaacgg cgtcgagctg agagacaatc agctcgtggt 1560 cccctccgag ggactgtatc tgatttactc ccaggtcctg tttaagggac agggatgccc 1620 tagcacacac gtcctgctga cccacaccat tagcaggatc gctgtgtcct accaaaccaa 1680 agtgaatctg ctgtccgcta tcaaaagccc ttgccaaaga gaaacccctg agggagccga 1740 agccaaaccc tggtacgaac ccatttacct cggcggagtg tttcagctgg agaaaggcga 1800 tagactcagc gctgagatta acaggcccga ttacctcgac tttgccgaaa gcggacaggt 1860 ctactttggc at tatcgctc tgtaaatcga ttcgtacgtc gacatcgagt gatgggtggc 1920 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca 1980 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata 2040 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag aca acctgta 2100 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg 2160 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat 2220 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca 2280 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc 2340 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa 2400 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac 2460 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt 2520 agatgcctgt tgaattggcg cgccggcctc cgcgccgggt tttggcgcct cccgcgggcg 2580 cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc 2640 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc t tagaacccc 2700 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct 2760 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat 2820 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag 2880 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatc Gctgt gatcgtcact 2940 tggtgagtag cgggctgctg ggctgggtac gtgcgctcgg ggttggcgag tgtgt tttgt 3000 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa • 118 - ttttcagtgt 3060

201207108 tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgagcta gcgccgccac catgggccct aaaaagaagc gtaaagtcgc ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac gagtacggtg gggaattcga gatgcctgtg gacaggatcc tggaggcaga gcttgctgtg gaacagaaga gtgaccaggg cgttgagggt cctgggggaa ccgggggtag cggcagcagc ccaaatgacc ctgtgactaa catctgtcag gcagctgaca aacagctatt cacgcttgtt gagtgggcga agaggatccc acacttttcc tccttgcctc tggatgatca ggtcatattg ctgcgggcag gctggaatga actcctcatt gcctcctttt cacaccgatc cattgatgtt cgagatggca tcctccttgc cacaggtctt cacgtgcacc gcaactcagc ccattcagca ggagtaggag ccatctttga tcgggtgctg acagagctag tgtccaaaat gcgtgacatg aggatggaca agacagagct tggctgcctg agggcaatca ttctgtttaa tccagaggtg aggggtttga aatccgccca ggaagttgaa cttctacgtg aaaaagtata tgccgctttg gaagaatata ctagaacaac acatcccgat gaaccaggaa gatttgcaaa acttttgctt cgtctgcctt ctttacgttc cataggcctt aagtgtttgg agcatttgtt tttctttcgc cttattggag atgttccaat tgatacgttc ctgatggaga tgcttgaatc accttctgat tcataatcta gcctagcccc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc tgaagacaaa ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatctct aggcgccacc atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc gccggaattc ccggggatcc ggcctgagtg cgtagtaccc gagactcagt gcgccatgaa gcggaaagag aagaaagcac agaaggagaa ggacaaactg cctgtcagca cgacgacggt ggacgaccac atgccgccca 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 -119 201207108 ttatgcagtg tgaacctcca cctcctgaag cagcaaggat tcacgaagtg gtcccaaggt 5340 ttctctccga caagctgttg gtgacaaacc ggcagaaaaa catcccccag ttgacagcca 5400 accagcagtt ccttatcgcc aggctcatct ggtaccagga cgggtacgag Cagccttctg 5460 atgaagattt gaagaggatt acgcagacgt ggcagcaagc ggacgatgaa aacgaagagt 5520 cggacactcc cttccgccag atcacagaga tgactatcct cacggtccaa cttat cgtgg 5580 agttcgcgaa gggattgcca gggttcgcca agatctcgca gcctgatcaa attacgctgc 5640 ttaaggcttg ctcaagtgag gtaatgatgc tccgagtcgc gcgacgatac gatgcggcct 5700 cagacagtat tctgttcgcg aacaaccaag cgtacactcg cgacaactac cgcaaggctg 5760 gcatggccga ggtcatcgag gatctactgc acttctgccg gtgcatgtac tctatggcgt 5820 tggacaacat ccattacgcg ctgctcacgg ctgtcgtcat cttttctgac cggccagggt 5880 tggagcagcc gcaactggtg gaagagatcc agcggtacta cctgaatacg ctccgcatct 5940 atatcctgaa ccagctgagc gggtcggcgc gttcgtccgt catatacggc aagatcctct 6000 caatcctctc tgagctacgc acgctcggca tgcaaaactc caacatgtgc atctccctca 6060 agctcaagaa cagaaagctg ccgcctttcc tcgaggagat ctgggatgtg gcggacatgt 6120 cgcacaccca accgccgcct atcctcgagt cccccacgaa tctctaggcg gcctctagag 6180 cggccgccac cgcggggaga tccagacatg ataagataca ttgatgagtt tggacaaacc 6240 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 6300 11 tgtaacca tattgcttta ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg 6360 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaa cct ctacaaatgt 6420 ggtatggctg attatgatcc ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct 6480 gacacatgca gctcccggag acggtcacag ct tgtctgta agcggatgcc gggagcagac 6540 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgaggtcga 6780 agcgatgaag atacagattg 6600 ctctagtccc cgcggtggca gatctggaag gtgctgaggt acgatgagac ccgcaccagg 6660 tgcagaccct gcgagtgtgg cggtaaacat attaggaacc agcctgtgat gctggatgtg 6720 accgaggagc tgaggcccga tcacttggtg ctggcctgca cccgcgctga gtttggctct aggtactgaa atgtgtgggc gtggcttaag ggtgggaaag 6840 aatatataag gtgggggtct tatgtagttt tgtatctgtt ttgcagcagc cgccgccgcc 6900 atgagcacca actcgtttga tggaagcatt gtgagctcat atttgacaac gcgcatgccc 6960 ccatgggccg gggtgcgtca gaatgtgatg ggctccagca ttgatggtcg ccccgtcctg 7020 cccgcaaact ctactacctt gacctacgag accgtgtctg gaacgccgtt ggagactgca 7080 gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac tgactttgct 7140 ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga tgacaagttg 7200 acggctcttt tggcacaatt ggattctttg acccgggaa c ttaatgtcgt ttctcagcag 7260 ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt 7320 taaaacataa ataaaaaacc agactctgtt tggatttgga tcaagcaagt gtcttgctgt 7380 ctttatttag gggttttgcg cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg 7440 gtcctgtgta ttttttccag gacgtggtaa aggtgactct ggatgttcag atacatgggc 7500 ataagcccgt ctctggggtg gaggtagcac cactgcagag cttcatgctg -120 - cggggtggtg 7560

C

201207108 ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt agcaagctga ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg gatgggtgca tacgtgggga tatgagatgc atcttggact gtatttttag gttggctatg ttcccagcca tatccctccg gggattcatg ttgtgcagaa ccaccagcac agtgtatccg gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa cttggagacg cccttgtgac ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg gcggcggcct gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg agatcgtcat aggccatttt tacaaagcgc gggcggaggg tgccagactg cggtataatg gttccatccg gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtaggg gagatcagct gggaagaaag caggttcctg agcagctgcg acttaccgca gccggtgggc ccgtaaatca cacctattac cggctgcaac tggtagttaa gagagctgca gctgccgtca tccctgagca ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc aaatccgcca gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc aggcggtccc acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcacg aggggcagtg cagacttttg agggcgtaga gcttgggcgc gagaaatacc gattccgggg agtaggcatc cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct ctggccgttc ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cagacttgag aggcctgtcc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 9720 9780 -121 201207108 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 9840 gttcagcccg accgctgcgc cttaiccggt aactatcgtc ttgagtccaa cccggtaaga 9900 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc Gaggtatgta 9960 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 10020 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctctt ga 10080 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 10140 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 10200 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 10260 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 10320 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 10380 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 10440 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 10500 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 10560 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 10620 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt 10680 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 10740 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 10800 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 10860 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 10920 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact 10980 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 11040 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 11100 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 11160 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 11220 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaa 11276 &lt; 210 &gt; 55 &lt; 211 &gt; 11120 &lt; 212 &gt; Wk &lt; 213 &gt; artificial sequence &lt; 220> &lt; 223 &gt; 5PolyDNA_from_WN-43326 &lt; 400 &gt; 55 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 60 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaattaag ctagcatcat caataatata ccttattttg gattgaagcc Aatatgataa 180 tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag 240 tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa 300 aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta 360 ggc Ggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa 420 actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg 480 -122 - 201207108

tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 780 tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg 1020 accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca 1080 catcccctga caagctgcca ggcaggttct cttcctctca catactgacc cacggctcca 1140 ccctctctcc cctggaaagg acaccgctag cgccaccatg tatagaatgc agctcctgtc 1200 ctgcattgcc ctgagcctcg ccctcgtgac aaactccgcc cctaccagcg gatccgtgag 1260 aagcagcagc aggaccccta gcgataagcc tgtggctcac gtcgtcgcta accctcaggc 1320 cgagggccag ctccagtggc tgaatagaag ggccaatgcc ctgctcgcca acggcgtcga 1380 gctgagagac aatcagctcg tggtcccctc cgagggactg tatctgattt actcccaggt 1440 cctgtttaag ggacagggat gccctagcac acacgtcctg ctgacccaca ccattagcag 1500 gatcgctgtg tcctaccaaa ccaaagtgaa tctgctgtcc gctatcaaaa gcccttgcca 1560 aagagaaacc cctgagggag ccgaagccaa accctggtac gaacccattt acctcggcgg 1620 agtgtttcag ctggagaaag gcgatagact cagcgctgag attaacaggc ccgattacct 1680 cgactttgcc gaaagcggac aggtctactt tggcattatc gctctgtaaa tcgattcgta 1740 cgtcgacatc gagtgatggg tggcatccct gtgacccctc cccagtgcct ctcctggccc 1800 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt 1860 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg 1920 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg 1980 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc 2040 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt 2100 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatct c 2160 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc 2220 ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc 2280 ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc 2340 tctcatgcgt tgggtccact cagtagatgc ctgttgaatt ggcgcgccgg cctccgcgcc 2400 gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc cattttagga cgggacttgg acgtcagacg 2460 aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct 2520 cataagactc ggccttagaa ccccagtatc agcagaagga 2580 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc 2640 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg 2700 .123 - 201207108 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt 2760 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg gtacgtgcgc 2820 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt 2880 gggtcaatat gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt 2940 ttggcttttt tgttagacga gctagcgccg Ccacat ggg ccctaaaaag aagcgtaaag 3000 tcgccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg 3060 cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt 3120 ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg 3180 acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtggggaat 3240 tcgagatgcc tgtggacagg atcctggagg cagagcttgc tgtggaacag aagagtgacc 3300 agggcgttga gggtcctggg ggaaccgggg gtagcggcag cagcccaaat gaccctgtga 3360 ctaacatctg tcaggcagct gacaaacagc tattcacgct tgttgagtgg gcgaagagga 3420 tcccacactt ttcctccttg cctctggatg atcaggtcat attgctgcgg gcaggctgga 3480 atgaactcct cattgcctcc ttttcacacc gatccattga tgttcgagat ggcatcctcc 3540 ttgccacagg tcttcacgtg caccgcaact cagcccattc agcaggagta ggagccatct 3600 ttgatcgggt gctgacagag ctagtgtcca aaatgcgtga catgaggatg gacaagacag 3660 agcttggctg cctgagggca atcattctgt ttaatccaga ggtgaggggt ttgaaatccg 3720 cccaggaagt tgaacttcta cgtgaaaaag tatatgccgc tttggaagaa tatactagaa 3780 caacacatcc cgatgaacca ggaagatttg caaaactttt gcttcgtctg ccttctttac 3840 gttccatagg ccttaagtgt ttggagcatt tgtttttctt tcgccttatt ggagatgttc 3900 caattgatac gttcctgatg gagatgcttg aatcaccttc tgattcataa tctagcctag 3960 cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg 4020 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc 4080 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa 4140 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga 4200 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc 4260 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc 4320 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac 4380 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg 4440 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg 4500 gggacgtggt tttcctttga aaaacacgat ctctaggcgc caccatgaag ctactgtct t 4560 ctatcgaaca agcatgcgat atttgccgac t taaaaagct caagtgctcc aaagaaaaac 4620 cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa 4680 ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa agactggaac 4740 agctatttct actgattttt cctcgagaag accttgacat gattttgaaa atggattctt 4800 tacaggatat aaaagcattg t taacaggat tatttgtaca agataatgtg aataaagatg 4860 ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata 4920 gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga • 124 cagttgactg 4980

201207108 tatcgccgga attcccgggg atccggcctg agtgcgtagt acccgagact cagtgcgcca tgaagcggaa agagaagaaa gcacagaagg agaaggacaa actgcctgtc agcacgacga cggtggacga ccacatgccg cccattatgc agtgtgaacc tccacctcct gaagcagcaa ggattcacga agtggtccca aggtttctct ccgacaagct gttggtgaca aaccggcaga aaaacatccc ccagttgaca gccaaccagc agttccttat cgccaggctc atctggtacc aggacgggta cgagcagcct tcigatgaag atttgaagag gattacgcag acgtggcagc aagcggacga tgaaaacgaa gagtcggaca ctcccttccg ccagatcaca gagatgacta tcctcacggt ccaacttatc gtggagttcg cgaagggatt gccagggttc gccaagatct cgcagcctga tcaaattacg ctgcttaagg cttgctcaag tgaggtaatg atgctccgag tcgcgcgacg atacgatgcg gcctcagaca gtattctgtt cgcgaacaac caagcgtaca ctcgcgacaa ctaccgcaag gctggcatgg ccgaggtcat cgaggatcta ctgcacttct gccggtgcat gtactctatg gcgttggaca acatccatta cgcgctgctc acggctgtcg tcatcttttc tgaccggcca gggttggagc agccgcaact ggtggaagag atccagcggt actacctgaa tacgctccgc atctatatcc tgaaccagct gagcgggtcg gcgcgttcgt ccgtcatata cggcaagatc ctctcaatcc tctctgagct acgcacgctc ggcatgcaaa actccaacat gtgcatctcc ctcaagctca agaacagaaa gctgccgcct ttcctcgagg agatctggga tgtggcggac atgtcgcaca cccaaccgcc gcctatcctc gagtccccca cgaatctcta ggcggcctct agagcggccg ccaccgcggg gagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgagg tcgactctag tccccgcggt ggcagatctg gaaggtgctg aggtacgatg agacccgcac caggtgcaga ccctgcgagt gtggcggtaa acatattagg aaccagcctg tgatgctgga tgtgaccgag gagctgaggc ccgatcactt ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat gaagatacag attgaggtac tgaaatgtgt gggcgtggct taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc cgcgggattg tgactgactt tgctttcctg agcccgcttg caagcagtgc agcttcccgt tcatccgccc gcgatgacaa gttgacggct cttttggcac aattggattc tttgacccgg gaacttaatg tcgtttcica gcagctgttg gatctgcgcc agcaggtttc tgccctgaag gcttcctccc ctcccaatgc ggtttaaaac ataaataaaa aaccagactc tgtttggatt 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 -125 - 201207108 tggatcaagc aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg gtaggcccgg 7260 gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg gtaaaggtga 7320 ctctggatgt tcagatacat gggcataagc ccgtctctgg Ggtggaggta gcaccactgc 7380 agagcttcat gctgcggggt ggtgttgtag atgatccagt cgtagcagga gcgctgggcg 7440 tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc ctt ggtgtaa 7500 gtgtttacaa agcggttaag ctgggatggg tgcatacgtg gggatatgag atgcatcttg 7560 gactgtattt ttaggttggc tccggggatt tatgttccca gccatatccc actgcggtat aatggttcca tccggcccag catgttgtgc 7620 agaaccacca gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag cttagaagga 7680 aatgcgtgga agaacttgga gacgcccttg tgacctccaa gattttccat gcattcgtcc 7740 ataatgatgg caatgggccc acgggcggcg gcctgggcga agatatttct gggatcacta 7800 acgtcatagt tgtgttccag gatgagatcg tcataggcca tttttacaaa gcgcgggcgg 7860 agggtgccag gggcgtagtt accctcacag 7920 atttgcattt cccacgcttt gagttcagat ggggggatca tgtctacctg cggggcgatg 7980 aagaaaacgg tttccggggt aggggagatc agctgggaag aaagcaggtt cctgagcagc 8040 tgcgacttac cgcagccggt gggcccgtaa atcacaccta ttaccggctg caactggtag 8100 ttaagagagc tgcagctgcc gtcatccctg agcagggggg ccacttcgtt aagcatgtcc 8160 ctgactcgca tgttttccct gaccaaatcc gccagaaggc gctcgccgcc cagcgatagc 8220 agttcttgca aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt aggcatgctt 8280 ttgagcgttt gaccaagcag ttccaggcgg tcccacagct cggtca cctg ctctacggca 8340 tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg tacggcagta 8400 gtcggtgctc gtccagacgg gccagggtca tgtctttcca cgggcgcagg gtcctcgtca 8460 gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc agggtgcgct 8520 tgaggctggt cctgctggtg ctgaagcgct gccggtcttc gccctgcgcg tcggccaggt 8580 agcat t tgac catggtgtca tagtccagcc cctccgcggc gtggcccttg gcgcgcagct 8640 tgcccttgga ggaggcgccg cacgaggggc agtgcagact tttgagggcg tagagcttgg 8700 gcgcgagaaa taccgattcc ggggagtagg catccgcgcc gcaggccccg cagacggtct 8760 cgcattccac gagccaggtg agctctggcc gttcggggtc aaaaaccagg tttcccccat 8820 gctttttgat gcgtttctta cctctggttt ccatgagccg gtgtccacgc tcggtgacga 8880 aaaggctgtc cgtgtccccg tatacagact tgagaggcct gtcctcgacc gatgcccttg 8940 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca 9000 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc 9060 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta 9120 ttcggaatct tgcacgccct cgctcaagcc ttcgtca ctg gtcccgccac caaacgtttc 9180 ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg 9240 gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc 9300 atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga 9360 cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 9420 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca -126 - gaggtggcga 9480 201207108

C

aacccgacag cctgttccga gcgctttctc ctgggctgtg cgtcttgagt aggattagca tacggctaca ggaaaaagag tttgtttgca ttttctacgg agattatcaa atctaaagta cctatctcag ataactacga ccacgctcac agaagtggtc agagtaagta gtggtgtcac cgagt tacat gttgtcagaa tctcttactg tcattctgag aataccgcgc cgaaaactct cccaactgat aggcaaaatg ttcctttttc tttgaatgta gactataaag ccctgccgct atagctcacg tgcacgaacc ccaacccggt gagcgaggta ctagaaggac ttggtagctc agcagcagat ggtctgacgc aaaggatctt tatatgagta cgatctgtct tacgggaggg cggctccaga ctgcaacttt gttcgccagt gctcgtcgtt gatcccccat gtaagttggc tcatgccatc aatagtgtat cacatagcag caaggatctt cttcagcatc ccgcaaaaaa aatattattg tttagaaaaa ataccaggcg taccggatac ctgtaggtat ccccgttcag aagacacgac tgtaggcggt agtatttggt ttgatccggc tacgcgcaga tcagtggaac cacctagatc aacttggtct atttcgttca cttaccatct tttatcagca atccgcctcc taatagtttg tggtatggct gttgtgcaaa cgcagtgtta cgtaagatgc gcggcgaccg aactttaaaa accgctgttg ttttactttc gggaataagg aagcatttat tttccccctg ctgtccgcct ctcagttcgg cccgaccgct ttatcgccac gctacagagt atctgcgctc aaacaaacc a aaaaaaggat gaaaactcac cttttaaatt gacagttacc tccatagttg ggccccagtg ataaaccagc atccagtcta cgcaacgttg tcattcagct aaagcggtta tcactcatgg ttttctgtga agttgctctt gtgctcatca agatccagtt accagcgttt gcgacacgga cagggttatt gaagctccct ttctcccttc tgtaggtcgt gcgccttatc tggcagcagc tcttgaagtg tgctgaagcc ccgctggtag ctcaagaaga gttaagggat aaaaatgaag aatgct taat cctgactccc ctgcaatgat cagccggaag ttaattgttg ttgccattgc ccggttccca gctccttcgg ttatggcagc ctggtgagta gcccggcgtc ttggaaaacg cgatgtaacc ctgggtgagc aatgttgaat gtctcatgag cgtgcgctct gggaagcgtg tcgctccaag cggtaactat cactggtaac gtggcctaac agttaccttc cggtggtttt tcct ttgatc tttggtcatg ttttaaatca cagtgaggca cgtcgtgtag accgcgagac ggccgagcgc ccgggaagct tgcaggcatc acgatcaagg tcctccgatc actgcataat ctcaaccaag aacacgggat ttcttcgggg cactcgtgca aaaaacagga actcatactc cggatacata 9540 9600 9660 9720 9780 9840 9900 9960 10020 10080 10140 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 11120 &lt;210> 56 &lt;211&gt; 11286 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; PolyI ^ A_froin_WN-43327 5U2 &lt; 400 &gt; 56 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag 60 120 180 240 -127 - 201207108 tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa 300 aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaatttteg cgcggtttta 360 ggcggatgtt gtagtaaatt tgggcgtaac egagtaagat ttggccattt tcgcgggaaa 420 actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg 480 tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa Taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtc c teegagegga gtactgtcct ccgagcggag 780 tactgtcctc cgagcggagt actgtcctcc gageggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc aegegtegaa gaaggtgagt aatcttaaca 1020 tgctcttttt tttttttttt gctaatccct tttgtgtgct gatgttagga tgacatttac 1080 aacaaatgtt tgttcctgac aggaaaaacc ttgctgggta ccttcgttgc cggacacttc 1140 ttgtcctcta ctttggaaaa aaggaattga gagccgctag cgccaccatg agcactgaaa 1200 gcatgatccg ggacgtggag ctggccgagg aagccctccc caagaaaacc ggcggccccc 1260 aggggagcag aagatgtttg ttcctgagcc tgttttcctt cctgatcgtg gcaggcgcta 1320 ccaccctgtt ctgcctgctg cactttggag tgatcggccc ccagagggag gagttcccca 1380 gggacctctc tctaatcagc cctctggccc aggeaggate egteagatea tettetegaa 1440 ccccgagtga caagcctgta gcccatgttg tagcaaaccc tcaagccgag ggccagctcc 1500 agtggctgaa ccgccgggcc aatgccctgc tcgccaacgg cgtcgagctg agagataacc 1560 agctggtggt gccatcagag ggcct gtacc tcatctactc ccaggtcctg ttcaagggcc 1620 aaggctgccc ctccacccat gtgctcctca cccacaccat cagccgcatc geegtgaget 1680 accagaccaa ggtcaacctc ctctctgcca tcaagagccc ctgccagagg gagaccccag 1740 agggggccga ggccaagccc tggtatgagc ccatctacct cggcggggtg ttccagctgg 1800 agaagggtga ccgactcagc gctgagatca atagacccga ctatctcgac tttgeegaga 1860 gcggccaggt gtactttggg atcattgccc tgtgaatcga ttcgtacgtc gacatcgagt 1920 gatgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact 1980 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg 2040 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag 2100 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct 2160 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag 2220 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga 2280 cggggtttca ccatatiggc caggctggtc tccaactcct aatctcaggt gatctaccca 2340 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt 2400 ctgattttaa aataacta Ta ccagcaggag gacgtccaga cacagcatag gctacctggc 2460 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg 2520 -128 - Ο

201207108 tccactcagt agatgcctgt tgaattggcg cgccggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctgggtac gtgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgagcta gcgccgccac catgggccct aaaaagaagc gtaaagtcgc ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac gagtacggtg gggaattcga gatgcctgtg gacaggatcc tggaggcaga gcttgctgtg gaacagaaga gtgaccaggg cgttgagggt cctgggggaa ccgggggtag cggcagcagc ccaaatgacc ctgtgactaa catctgtcag gcagctgaca aacagctatt cacgcttgtt gagtgggcga agaggatccc acacttttcc tccttgcctc tggatgatca ggtcatattg ctgcgggcag gctggaatga actcctcatt gcctcctttt cacaccgatc cattgatgtt cgagatggca tcctccttgc cacaggtctt cacgtgcacc gcaactcagc ccattcagca ggagtaggag ccatctttga tcgggtgctg acagagctag tgtccaaaat gcgtgacatg aggatggaca agacagagct tggctgcctg agggcaatca ttctgtttaa tccagaggtg aggggtttga aatccgccca ggaagttgaa cttctacgtg aaaaagtata tgccgctttg gaagaatata ctagaacaac acatcccgat gaaccaggaa gatttgcaaa acttttgctt cgtctgcctt ctttacgttc cataggcctt aagtgtttgg agcatttgtt tttctttcgc cttattggag atgttccaat tgatacgttc ctgatggaga tgcttgaatc accttctgat tcataatcta gcctagcccc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatctct aggcgccacc atgaagctac tgtcttctat cgaacaagca 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 -129 - 201207108 tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag 4800 tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact 4860 agggcacatc tgacagaagt ggaatcaagg ctagaaagac Tggaacagct atttctactg 4920 atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca ggatataaaa 4980 gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt cac agataga 5040 ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat aagtgcgaca 5100 tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc gccggaattc 5160 ccggggatcc ggcctgagtg ggtaccagga cgggtacgag cgtagtaccc gagactcagt gcgccatgaa gcggaaagag 5220 aagaaagcac agaaggagaa ggacaaactg cctgtcagca cgacgacggt ggacgaccac 5280 atgccgccca ttatgcagtg tgaacctcca cctcctgaag cagcaaggat tcacgaagtg 5340 gtcccaaggt ttctctccga caagctgttg gtgacaaacc ggcagaaaaa catcccccag 5400 ttgacagcca accagcagtt ccttatcgcc aggctcatct 5460 cagccttctg atgaagattt gaagaggatt acgcagacgt ggcagcaagc ggacgatgaa 5520 aacgaagagt cggacactcc cttccgccag atcacagaga tgactatcct cacggtccaa 5580 cttatcgtgg agttcgcgaa gggattgcca gggttcgcca agatctcgca gcctgatcaa 5640 attacgctgc ttaaggcttg ctcaagtgag gtaatgatgc tccgagtcgc gcgacgatac 5700 gatgcggcct cagacagtat tctgttcgcg aacaaccaag cgtacactcg cgacaactac 5760 cgcaaggctg gcatggccga ggtcatcgag gatctactgc acttctgccg gtgcatgtac 5820 tctatggcgt tggacaacat ccattacgcg ctgctcacgg ctgtcg tcat cttttctgac 5880 cggccagggt tggagcagcc agcggtacta gcaactggtg gaagagatcc tggacaaacc acaactagaa tgcagtgaaa cctgaatacg 5940 ctccgcatct atatcctgaa ccagctgagc gggtcggcgc gttcgtccgt catatacggc 6000 aagatcctct caatcctctc tgagctacgc acgctcggca tgcaaaactc caacatgtgc 6060 atctccctca agctcaagaa cagaaagctg ccgcctttcc tcgaggagat ctgggatgtg 6120 gcggacatgt cgcacaccca accgccgcct atcctcgagt cccccacgaa tctctaggcg 6180 gcctctagag cggccgccac cgcggggaga tccagacatg ataagataca ttgatgagtt 6240 aaaatgcttt atttgtgaaa tttgtgatgc 6300 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 6360 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct 6420 ctacaaatgt ggtatggctg attatgatcc ggctgcctcg cgcgtttcgg tgatgacggt 6480 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc 6540 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc 6600 atgaggtcga ctctagtccc cgcggtggca gatctggaag gtgctgaggt acgatgagac 6660 ccgcaccagg tgcagaccct gcgagtgtgg cggtaaaca t attaggaacc agcctgtgat 6720 gctggatgtg accgaggagc tgaggcccga tcacttggtg ctggcctgca cccgcgctga 6780 gtttggctct agcgatgaag atacagattg aggtactgaa atgtgtgggc gtggcttaag 6840 ggtgggaaag aatatataag gtgggggtct tatgtagttt tgtatctgtt ttgcagcagc 6900 cgccgccgcc atgagcacca actcgtttga tggaagcatt gtgagctcat atttgacaac 6960 gcgcatgccc ccatgggccg gggtgcgtca gaatgtgatg ggctccagca -130 · ttgatggtcg 7020

201207108 ccccgtcctg cccgcaaact ctactacctt gacctacgag accgtgtctg gaacgccgtt ggagactgca gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac t £ actttgct ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga tgacaagttg acggctcttt tggcacaatt ggattctttg acccgggaac ttaatgtcgt ttctcagcag ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt taaaacataa ataaaaaacc agactctgtt tggatttgga tcaagcaagt gtcttgctgt ctttatttag gggttttgcg cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg gtcctgtgta ttttttccag gacgtggtaa aggtgactct ggatgttcag atacatgggc ataagcccgt ctctggggtg gaggtagcac cactgcagag cttcatgctg cggggtggtg ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt agcaagctga ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg gatgggtgca tacgtgggga tatgagatgc atcttggact gtatttttag gttggctatg ttcccagcca tatccctccg gggattcatg ttgtgcagaa ccaccagcac agtgtatccg gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa cttggagacg cccttgtgac ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg gcggcggcct gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg agatcgtcat aggccatttt acttaccgca gccggtgggc ccgtaaatca cacctattac cggctgcaac tggtagttaa gagagctgca gctgccgtca tccctgagca ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc aaatccgcca gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt ttcaacggtt tacaaagcgc gggcggaggg tgccagactg cggtataatg gttccatccg gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtaggg gagatcagct gggaagaaag caggttcctg agcagctgcg tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc aggcggtccc acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcac g aggggcagtg cagacttttg agggcgtaga gcttgggcgc gagaaatacc gattccgggg agtaggcatc cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct ctggccgttc ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cagacttgag aggcctgtcc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 -131 · 201207108 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca 9300 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc 9360 cattatcgcc ggcatggcgg ccgacgcgct Gggctacgtc ttgctggcgt tcgcgacgcg 9420 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc 9480 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaa ggcca 9540 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 9600 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 9660 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 9720 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 9780 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 9840 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 9900 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 9960 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 10020 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 10080 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 10140 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 10200 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 10260 gatcttcacc tagatccttt taaattaaaa aigaagtttt aaatcaatct aaagtatata 10320 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctca gcgat 10380 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 10440 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 10500 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 10560 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 10620 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc 10680 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 10740 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 10800 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 10860 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 10920 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca 10980 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 11040 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 11100 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 11160 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactctt Cc tttttcaata 11220 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 11280 gaaaaa 31286 &lt;210&gt; 57 &lt;211&gt; 11286 &lt;2)2&gt; DNA &lt;213&gt; artificial sequence -132 - f

201207108 &lt; 220> &lt; 223 &gt; PolyEWA_froni_WN-43328 &lt; 400 &gt; 57 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg Agcggagtac tgtcc tccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag ctggtgtgtg agctcatctt cctgtagatc acgcgtcgaa gaaggtgagt aatcttaaca tgctcttttt tttttttttt gctaatccct tttgtgtgct gatgttagga tgacatttac aacaaatgtt tgttcctgac aggaaaaacc ttgctgggta ccttcgttgc cggacacttc ttgtcctcta ctttggaaaa aaggaattga gagccgctag cgccaccatg tccaccgaaa gcatgatccg ggacgtggag ctggccgagg aagccctgcc taagaaaacc ggaggccctc agggaagcag gagatgtctg tttctgtccc tgtttagctt tctgattgtg gctggcgcta ccacactgtt ttgcctcctg catttcggag tgattggccc tcagagggag gagttcccta gagacctgtc cctgattagc cctctggctc aggctggatc cgtgagaagc agcagcagga cccctagcga taagcctgtg gctcacgtcg tcgctaaccc tcaggccgag ggccagctcc agtggctgaa tagaagggcc aatgccctgc tcgccaacgg cgtcgagctg agagacaatc agctcgtggt cccctccgag ggactgtatc tgatttactc ccaggtcctg tttaagggac agggatgccc tagcacacac gtcctgctga cccacaccat tagcaggatc gctgtgtcct accaaaccaa agtgaatctg ctgtccgcta tcaaaagccc ttgccaaaga gaaacccctg agggagccga agcc aaaccc tggtacgaac ccatttacct cggcggagtg tttcagctgg agaaaggcga tagactcagc gctgagatta acaggcccga ttacctcgac tttgccgaaa gcggacaggt ctactttggc attatcgctc tgtaaatcga ttcgtacgtc gacatcgagt gatgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 I860 1920 1980 2040 2100 -133 - 201207108 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct 2160 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag 2220 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga 2280 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt Gatctaccca 2340 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccct t ccctgtcctt 2400 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc 2460 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg 2520 tccactcagt agatgcctgt tgaattggcg cgccggcctc cgcgccgggt tttggcgcct 2580 cccgcgggcg cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag 2640 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcaia agactcggcc 2700 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca 2760 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct 2820 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg 2880 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt 2940 gatcgtcact tggtgagtag cgggctgctg ggctgggtac gtgcgctcgg ggttggcgag 3000 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa 3060 ttttcagtgt tagactagta aattgtccgc taaat tctgg ccgtttttgg cttttttgtt 3120 agacgagcta gcgccgccac catgggccct aaaaagaagc gtaaagtcgc ccccccgacc 3180 gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc 3240 gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga 3300 tt tacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag 3360 cagatgttta ccgatgccct tggaattgac gagtacggtg gggaattcga gatgcctgtg 3420 gacaggatcc tggaggcaga gcttgctgtg gaacagaaga gtgaccaggg cgttgagggt 3480 cctgggggaa ccgggggtag cggcagcagc ccaaatgacc ctgtgactaa catctgtcag 3540 gcagctgaca aacagctat t cacgcttgt t gagtgggcga agaggatccc acacttttcc 3600 tccttgcctc tggatgatca ggtcatattg ctgcgggcag gctggaatga actcctcatt 3660 gcctcctttt cacaccgatc cattgatgtt cgagatggca tcctccttgc cacaggtctt 3720 cacgtgcacc gcaactcagc ccattcagca ggagtaggag ccatctttga tcgggtgctg 3780 acagagctag tgtccaaaat gcgtgacatg aggatggaca agacagagct tggctgcctg 3840 agggcaatca ttctgtttaa tccagaggtg aggggtttga aatccgccca ggaagttgaa 3900 cttctacgtg aaaaagtata tgccgctttg gaagaatata ctagaacaac acatcccgat 3960 gaaccaggaa gatttgcaaa acttttgctt cgtctgcctt ctttacgttc cataggcctt 4020 aagtgtttgg agcatttgtt tttctttcgc cttattggag atgttccaat tgatacgttc 4080 ctgatggaga tgcttgaatc accttctgat tcataatcta gcctagcccc cctctccct c 4140 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgittgtcta 4200 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc 4260 tgtcttcttg acgagcat tc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct 4320 gttgaatgtc gtgaaggaag cagttcctct ggaagcttcl tgaagacaaa -134 - caacgtctgt 4380

201207108 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatctct aggcgccacc atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc gccggaattc ccggggatcc ggcctgagtg cgtagtaccc gagactcagt gcgccatgaa gcggaaagag aagaaagcac agaaggagaa ggacaaactg cctgtcagca cgacgacggt ggacgaccac atgccgccca ttatgcagtg tgaacctcca cctcctgaag cagcaaggat tcacgaagtg gtcccaaggt ttctctccga caagctgttg gtgacaaacc ggcagaaaaa catcccccag ttgacagcca accagcagtt ccttatcgcc aggctcatct ggtaccagga cgggtacgag cagccttctg atgaagattt gaagaggatt acgcagacgt ggcagcaagc ggacgatgaa aacgaagagt cggacactcc cttccgccag atcacagaga tgactatcct cacggtccaa cttatcgtgg agttcgcgaa gggattgcca gggttcgcca agatctcgca gcctgatcaa attacgctgc ttaaggcttg ctcaagtgag gtaatgatgc tccgagtcgc gcgacgatac gatgcggcct cagacagtat tctgttcgcg aacaaccaag cgtacactcg cgacaactac cgcaaggctg gcatggccga ggtcatcgag gatctactgc acttctgccg gtgcatgtac tctatggcgt tggacaacat ccattacgcg ctgctcacgg ctgtcgtcat cttttctgac cggccagggt tggagcagcc gcaactggtg gaagagatcc agcggtacta cctgaatacg ctccgcatct atatcctgaa ccagctgagc gggtcggcgc gttcgtccgt catatacggc aagatcctct caatcctctc tgagctacgc acgctcggca tgcaaaactc caacatgtgc tctctaggcg gcctctagag atctccctca agctcaagaa cagaaagctg ccgcctttcc tcgaggagat ctgggatgtg gcggacatgt cgcacaccca accgccgcct atcctcgagt cccccacgaa cggccgccac cgcggggaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6380 6240 6300 6360 6420 6480 6540 6600 135 - 201207108 atgaggtcga ctctagtccc cgcggtggca gatctggaag gtgctgaggt acgatgagac 6660 ccgcaccagg tgcagaccct gcgagtgtgg cggtaaacat attaggaacc agcctgtgat 6720 gctggatgtg accgaggagc tgaggcccga tcacttggtg ctggcctgca Cccgcgctga 6780 gtttggctct agcgatgaag atacagattg aggtactgaa atgtgtgggc gtggcttaag 6840 ggtgggaaag aatatataag gtgggggtct tatgtagttt tgtatctgtt ttgcagca gc 6900 cgccgccgcc atgagcacca actcgtttga tggaagcatt gtgagctcat atttgacaac 6960 gcgcatgccc ccatgggccg gggtgcgtca gaatgtgatg ggctccagca ttgatggtcg 7020 ccccgtcctg cccgcaaact ctactacctt gacctacgag accgtgtctg gaacgccgtt 7080 ggagactgca gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac 7140 tgactttgct ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga 7200 tgacaagttg acggctcttt tggcacaatt ggattctttg acccgggaac ttaatgtcgt 7260 ttctcagcag ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc 7320 caatgcggtt taaaacataa ataaaaaacc agactctgtt tggatttgga tcaagcaagt 7380 gtcttgctgt ctttatttag gggttttgcg cgcgcggtag gcccgggacc agcggtctcg 7440 gtcgttgagg gtcctgtgta ttttttccag gacgtggtaa aggtgactct ggatgttcag 7500 atacatgggc ataagcccgt ctctggggtg gaggtagcac cactgcagag cttcatgctg 7560 cggggtggtg ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat 7620 gtctttcagt agcaagctga ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg 7680 gttaagctgg gatgggtgca tacgtgggga tatgagatgc atcttggact gtatttttag 774 0 gttggctatg ttcccagcca tatccctccg gggattcatg ttgtgcagaa ccaccagcac 7800 agtgtatccg gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa 7860 cttggagacg cccttgtgac ctccaagatt ttccatgcat tcgtccataa tgatggcaat 7920 gggcccacgg gcggcggcct gggcgaagat atttctggga tcactaacgt catagttgtg 7980 ttccaggatg agatcgtcat aggccatttt tacaaagcgc gggcggaggg tgccagactg 8040 cggtataatg gttccatccg gcccaggggc gtagttaccc tcacagattt gcatttccca 8100 cgctttgagt tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc 8160 cggggtaggg gagatcagct gggaagaaag caggttcctg agcagctgcg acttaccgca 8220 gccggtgggc ccgtaaatca cacctattac cggctgcaac tggtagttaa gagagctgca 8280 gctgccgtca tccctgagca ggggggccac ttcgttaagc atgtccctga ctcgcatgtt 8340 ttccctgacc aaatccgcca gaaggcgctc gccgcccagc gatagcagtt cttgcaagga 8400 agcaaagttt ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc 8460 aagcagttcc aggcggtccc acagctcggt cacctgctct acggcatctc gatccagcat 8520 atctcctcgt ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc 8580 agac gggcca gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc 8640 acggtgaagg ggtgcgctcc gggctgcgcg ctggccaggg tgcgctigag gctggtcctg 8700 ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg 8760 gtgtcatagt ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag 8820 gcgccgcacg aggggcagtg cagacttttg agggcgtaga gcttgggcgc gagaaatacc 8880 -136 -

201207108 gattccgggg agtaggcatc cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct ctggccgttc ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cagacttgag aggcctgtcc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 9720 9780 9840 9900 9960 10020 10080 10140 10200 10260 10320 10380 10440 10500 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 • 137 - 201207108 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 11160 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 11220 ttattgaagc atttatcagg gttattgtct catgagcgga Tacatatttg aatgtattta 11280 gaaaaa 11286 &lt;210&gt; 58 &lt;211&gt; 11130 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt;; PolyDNA_from_WN-43329 &lt; 400 &gt; 58 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 60 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa 180 tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag 240 tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa 300 aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta 360 ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa 420 actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg 480 tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 780 Tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctc cg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtcgaa gaaggtgagt aatcttaaca 1020 tgctcttttt tttttttttt gctaatccct tttgtgtgct gatgttagga tgacatttac 1080 aacaaatgtt tgttcctgac aggaaaaacc ttgctgggta ccttcgttgc cggacacttc 1140 ttgtcctcta ctttggaaaa aaggaattga gagccgctag cgccaccatg tatagaatgc 1200 agctcctgtc ctgcattgcc ctgagcctcg ccctcgtgac aaactccgcc cctaccagcg 1260 gatccgtgag aagcagcagc aggaccccta gcgataagcc tgtggctcac gtcgtcgcta 1320 accctcaggc cgagggccag ctccagtggc tgaatagaag ggccaatgcc ctgctcgcca 1380 acggcgtcga gctgagagac aatcagctcg tggtcccctc cgagggactg tatctgattt 1440 actcccaggt cctgtttaag ggacagggat gccctagcac acacgtcctg ctgacccaca 1500 ccattagcag gatcgctgtg icctaccaaa ccaaagtgaa tctgctgtcc gctatcaaaa 1560 gcccttgcca aagagaaacc cctgagggag ccgaagccaa accctggtac gaacccattt 1620 acctcggcgg agtgtttcag ctggagaaag gcgatagact cagcgctgag attaac Aggc 1680 ccgattacct cgactttgcc gaaagcggac aggtctaclt tggcattatc gctctgtaaa 1740 -138 - 201207108

tcgattcgta ctcctggccc tgcatcattt tggagcaagg agctggagtg attctcctgc taatttttgt tcctaatctc accactgctc cagacacagc ggcactgtcc cctccgcgcc acgtcagacg gcccgctgct cgggacttgg aagtagtccc ttatataagg cggttcttgt gtacgtgcgc gtaatcattt ctggccgttt aagcgtaaag gaggacgtgg gacggggatt gatatggccg ggtggggaat aagagtgacc gaccctgtga gcgaagagga gcaggctgga ggcatcctcc ggagccatct gacaagacag ttgaaatccg tatactagaa ccttctttac ggagatgttc cgtcgacatc tggaagttgc tgtctgacta ggcaagttgg cagtggcaca ctcagcctcc ttttttggta aggtgatcta ccttccctgt ataggctacc tctcatgcgt gggttttggc aagggcgcag cataagactc gtgactctag ttctcggcga acgcgccggg ttgtggatcg tcggggttgg gggtcaatat ttggcttttt tcgccccccc cgatggcgca ccccgggtcc acttcgagtt tcgagatgcc agggcgttga ctaacatctg tcccacactt atgaactcct ttgccacagg ttgatcgggt agcttggctg cccaggaagt caacacatcc gttccatagg caattgatac gagtgatggg cactccagtg ggtgtccttc gaagacaacc atcttggctc cgagttgttg gagacggggt cccaccttgg ccttctgatt tggccatgcc tgggtccact gcctcccgcg cgagcgtcct ggccttagaa ggcactggtt ttctgcggag tgtggcacag ctgtgatcgt cgagtgtgtt gtaattttca tgttagacga gaccgatgtc tgccgacgcg gggatttacc tgagcagatg tgtggacagg gggtcctggg tcaggcagct ttcctccttg cattgcctcc tcttcacgtg gctgacagag cctgagggca tgaacttcta cgatgaacca ccttaagtgt gttcctgatg tggcatccct cccaccagcc tataatatta tgtagggcct actgcaatct ggattccagg ttcaccatat cctcccaaat ttaaaataac caaccggtgg cagtagatgc ggcgcccccc gatccttccg ccccagtatc ttctttccag ggatctccgt ctagttccgt cacttggtga ttgtgaagtt gtgttagact gctagcgccg agcctggggg ctagacgatt ccccacgact tttaccgatg atcctggagg ggaaccgggg gacaaacagc cctctggatg ttttcacacc caccgcaact ctagtgtcca atcattctgt cgtgaaaaag ggaagatttg ttggagcatt gagatgcttg gtgacccctc ttgtcctaat tggggtg £ ag gcggggtcta ccgcctcctg catgcatgac tggccaggct tgctgggatt tataccagca gacatttgag ctgttgaatt tcctcacggc cccggacgct agcagaagga agagcggaac ggggcggtga cgcagccggg gtagcgggct ttttaggcac agtaaattgt ccaccatggg acgagctcca tcgatctgga ccgcccccta cccttggaat cagagcttgc gtagcggcag tattcacgct atcaggtcat gatccattga cagcccattc aaatgcgtga ttaatccaga tatatgcc gc caaaactttt tgtttttctt aatcaccttc cccagtgcct aaaattaagt gggggtggta ttgggaacca ggttcaagcg caggctcagc ggtctccaac acaggcgtga ggaggacgtc ttgcttgctt ggcgcgccgg gagcgctgcc caggacagcg cattttagga aggcgaggaa acgccgatga atttgggtcg gctgggctgg cttttgaaat ccgctaaatt ccctaaaaag cttagacggc catgttgggg cggcgctctg tgacgagtac tgtggaacag cagcccaaat tgttgagtgg attgctgcgg tgttcgagat agcaggagta catgaggatg ggtgaggggt tttggaagaa gcttcgtctg tcgccttatt tgattcataa 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 -139 - 201207108 tctagcctag cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga 4020 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa 4080 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc Attcctaggg gtctttcccc 4140 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc 4200 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg 4 260 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca 4320 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag 4380 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct 4440 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc 4500 ccgaaccacg gggacgtggt tttcctttga aaaacacgat ctctaggcgc caccatgaag 4560 ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc 4620 aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc 4680 aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa 4740 agactggaac agctatttct actgattttt cctcgagaag accttgacat gattttgaaa 4800 atggat tct t tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg 4860 aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg 4920 agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga 4980 cagttgactg tatcgccgga attcccgggg atccggcctg agtgcgtagt acccgagact 5040 cagtgcgcca tgaagcggaa agagaagaaa gcacagaagg agaaggacaa ac tgcctgtc 5100 agcacgacga cggtggacga ccacatgccg cccattatgc agtgtgaacc tccacctcct 5160 gaagcagcaa ggattcacga agtggtccca aggtttctct ccgacaagct gttggtgaca 5220 aaccggcaga aaaacatccc ccagt tgaca gccaaccagc agttccttat cgccaggctc 5280 atctggtacc aggacgggta cgagcagcct tctgatgaag atttgaagag gattacgcag 5340 acgtggcagc aagcggacga tgaaaacgaa gagtcggaca ctcccttccg ccagatcaca 5400 gagatgacta tcctcacggt ccaacttatc gtggagttcg cgaagggatt gccagggttc 5460 gccaagatct cgcagcctga tcaaattacg ctgcttaagg cttgctcaag tgaggtaatg 5520 atgctccgag tcgcgcgacg atacgatgcg gcctcagaca gtattctgtt cgcgaacaac 5580 caagcgtaca ctcgcgacaa ctaccgcaag gctggcatgg ccgaggtcat cgaggatcta 5640 ctgcacttct gccggtgcat gtactctatg gcgttggaca acatccatta cgcgctgctc 5700 acggctgtcg tcatcttttc tgaccggcca gggttggagc agccgcaact ggtggaagag 5760 atccagcggt actacctgaa tacgctccgc atctatatcc tgaaccagct gagcgggtcg 5820 gcgcgttcgt ccgtcatata cggcaagatc ctctcaatcc tctctgagct acgcacgctc 5880 ggcatgcaaa actccaacat gtgcatctcc ctcaagctca agaa cagaaa gctgccgcct 5940 ttcctcgagg agatctggga tgtggcggac atgtcgcaca cccaaccgcc gcctatcctc 6000 gagtccccca cgaatctcta ggcggcctct agagcggccg ccaccgcggg gagatccaga 6060 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 6120 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 6180 acaagttaac aacaacaat t gcattcattt tatgtttcag gttcaggggg -140 · aggtgtggga 6240 201207108

ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atccggctgc 6300 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 6360 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 6420 gttggcgggt gtcggggcgc agccatgagg tcgactctag tccccgcggt ggcagatctg 6480 gaaggtgctg aggtacgatg agacccgcac caggtgcaga ccctgcgagt gtggcggtaa 6540 acatattagg aaccagcctg tgatgctgga tgtgaccgag gagctgaggc ccgatcactt 6600 ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat gaagatacag attgaggtac 6660 tgaaatgtgt gggcgtggct taagggtggg aaagaatata taaggtgggg gtcttatgta 6720 gttttgtatc tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag 6780 cattgtgagc tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt 6840 gatgggctcc agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta 6900 cgagaccgtg tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc 6960 agccaccgcc cgcgggattg tgactgactt tgctttcctg agcccgcttg caagcagtgc 7020 agcttcccgt tcatccgccc gcgatgacaa gttgacggct cttttggcac aattggatt c 7080 tttgacccgg gaacttaatg tcgtttctca gcagctgttg gatctgcgcc agcaggtttc 7140 tgccctgaag gcttcctccc ctcccaatgc ggtttaaaac ataaataaaa aaccagactc 7200 tgtttggatt tggatcaagc aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg 7260 gtaggcccgg gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg 7320 gtaaaggtga ctctggatgt tcagatacat gggcataagc ccgtctctgg ggtggaggta 7380 gcaccactgc agagcttcat gctgcggggt ggtgttgtag atgatccagt cgtagcagga 7440 gcgctgggcg tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc 7500 cttggtgtaa gtgtttacaa agcggttaag ctgggatggg tgcatacgtg gggatatgag 7560 atgcatcttg gactgtattt ttaggttggc tatgttccca gccatatccc tccggggatt 7620 catgttgtgc agaaccacca gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag 7680 cttagaagga aatgcgtgga agaacttgga gacgcccttg tgacctccaa gattttccat 7740 gcattcgtcc ataatgatgg caatgggccc acgggcggcg gcctgggcga agatatttct 7800 gggatcacta acgtcatagt tgtgttccag gatgagatcg tcataggcca tttttacaaa 7860 gcgcgggcgg agggtgccag actgcggtat aatggttcca tccggcccag g ggcgtagtt 7920 accctcacag atttgcattt cccacgcttt gagttcagat ggggggatca tgtctacctg 7980 cggggcgatg aagaaaacgg tttccggggt aggggagatc agctgggaag aaagcaggtt 8040 cctgagcagc tgcgacttac cgcagccggt gggcccgtaa atcacaccta ttaccggctg 8100 caactggtag ttaagagagc tgcagctgcc gtcatccctg agcagggggg ccacttcgtt 8160 aagcatgtcc ctgactcgca tgttttccct gaccaaatcc gccagaaggc gctcgccgcc 8220 cagcgatagc agttcttgca aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt 8280 aggcatgctt ttgagcgttt gaccaagcag ttccaggcgg tcccacagct cggtcacctg 8340 ctctacggca tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg 8400 tacg £ cagta gtcggtgctc gtccagacgg gccagggtca tgtctttcca cgggcgcagg 8460 -141 - 201207108 gtcctcgtca gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc 8520 agggtgcgct tgaggctggt cctgctggtg ctgaagcgct gccggtcttc gccctgcgcg 8580 tcggccaggt agcatttgac catggtgtca tagtccagcc cctccgcggc gtggcccttg 8640 gcgcgcagct tgcccttgga ggaggcgccg cacgaggggc agtgcagact tttgagggcg 8700 tagagcttgg Gcgcgagaaa taccgattc c ggggagtagg catccgcgcc gcaggccccg 8760 cagacggtct cgcattccac gagccaggtg agctctggcc gttcggggtc aaaaaccagg 8820 tttcccccat gctttttgat gcgtttctta cctctggttt ccatgagccg gtgtccacgc 8880 tcggtgacga aaaggctgtc cgtgtccccg tatacagact tgagaggcct gtcctcgacc 8940 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat 9000 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc 9060 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc 9120 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac 9180 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta 9240 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc 9300 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgaega 9360 ccatcaggga cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 9420 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 9480 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 9540 cgtgcgctct cctgttccga c cctgccgct taccggatac ctgtccgcct ttctcccttc 9600 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 9660 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 9720 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggeageage 9780 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tettgaagtg 9840 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 9900 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 9960 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 10020 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 10080 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 10140 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 10200 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 10260 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 10320 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cageeggaag 10380 ggccgagc gc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 10440 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 10500 tgcaggcatc gtggtgtcac gcicgtcgtt tggtatggct teattcagct ccggttccca 10560 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 10620 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 10680 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga • 142 - ctggtgagta 10740

C

201207108 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa &lt; 210 &gt; 59 &lt; 211 &gt; 11824 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; PolyDNA_from_WN-43533 &lt; 400 &gt; 59 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gatigaagcc aatatgataa tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt Gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaa c cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca catcccctga caagctgcca ggcaggttct cttcctctca catactgacc cacggctcca ccctctctcc cctggaaagg acaccgctag cgccaccatg agcactgaaa gcatgatccg ggacgtggag ctggccgagg aagccctccc caagaaaacc ggcggccccc aggggagcag aagatgtttg ttcctgagcc tgttttcctt cctgatcg tg gcaggcgcta ccaccctgtt ctgcctgctg cactttggag tgatcggccc ccagagggag gagttcccca gggacctctc tctaatcagc cctctggccc aggcaggatc cgtcagatca tcttctcgaa ccccgagtga caagcctgta gcccatgttg tagcaaaccc tcaagccgag ggccagctcc agtggctgaa 10800 10860 10920 10980 11040 11100 11130 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 143 - 201207108 ccgccgggcc aatgccctgc tcgccaacgg cgtcgagctg agagataacc agctggtggt 1560 gccatcagag ggcctgtacc tcatctactc ccaggtcctg ttcaagggcc aaggctgccc 1620 ctccacccat gtgctcctca cccacaccat cagccgcatc gccgtgagct accagaccaa 1680 ggtcaacctc ctctctgcca tcaagagccc ctgccagagg gagaccccag agggggccga 1740 ggccaagccc tggtatgagc ccatctacct cggcggggtg ttccagctgg agaagggtga 1800 ccgactcagc gctgagatca atagacccga ctatctcgac tttgccgaga gcggccaggt 1860 gtactttggg atcattgccc Tgtgaatcga ttacgtagcg gccgcgtcga cggaggacga 1920 acatccaacc ttcccaaacg cctcccctgc cccaatccct ttattacccc ctccttcaga 1980 caccctcaac ctcttctggc tcaaaaagag aa ttgggggc ttagggtcgg aacccaagct 2040 tagaacttta agcaacaaga ccaccacttc gaaacctggg attcaggaat gtgtggcctg 2100 cacagtgaag tgctggcaac cactaagaat tcaaactggg gcctccagaa ctcactgggg 2160 cctacagctt tgatccctga catctggaat ctggagacca gggagccttt ggttctggcc 2220 agaatgctgc aggacttgag aagacctcac ctagaaattg acacaagtgg accttaggcc 2280 ttcctctctc cagatgtttc cagacttcct tgagacacgg agcccagccc tccccatgga 2340 gccagctccc tctatttatg tttgcacttg tgattattta ttatttattt attatttatt 2400 tatttacaga tgaatgtatt tatttgggag accggggtat cctgggggac ccaatgtagg 2460 agctgccttg gctcagacat gttttccgtg aaaacggagc tgaacaatag gctgttccca 2520 tgtagccccc tggcctctgt gccttctttt gattatgttt tttaaaatat ttatctgatt 2580 aagttgtcta aacaatgctg atttggtgac caactgtcac tcattgctga gcctctgctc 2640 cccaggggag ttgtgtctgt aatcgcccta ctattcagtg gcgagaaata aagtttgctt 2700 agaaaagaaa catggtctcc ttcttggaat taattctgca tctgcctctt cttgtgggtg 2760 ggaagaagct ccctaagtcc tctctccaca ggctttaaga tccctcggac ccagtcccat 2820 ccttagactc ctagggccct ggaga cccta cataaacaaa gcccaacaga atattcccca 2880 tcccccagga aacaagagcc tgaacctaat tacctctccc tcagggcatg ggaatttcca 2940 actctgggaa ttccaatcct tgctgggaaa atcctgcagc tcaggtgaga tttccggctg 3000 ttgcagctgg ccagcagtcc ggagagagct ggagaggagc cgcattctca ggtacctgaa 3060 tcacacattt aaatcggtcc gcgtaccgcg ccggcctccg cgccgggttt tggcgcctcc 3120 cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg 3180 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt 3240 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact 3300 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc 3360 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc 3420 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga 3480 tcgtcacttg gtgagtagcg ggctgctggg ctgggtacgt gcgctcgggg ttggcgagtg 3540 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt 3600 ttcagtgtta gactagtaaa t tgtccgcta aat tctggcc gtttttggct tttttgttag 3660 acgagctagc gccgcc Acca tgggccctaa aaagaagcgt aaagtcgccc ccccgaccga 3720 tgtcagcctg ggggacgagc iccact taga cggcgaggac gtggcgatgg -144 - cgcatgccga 3780

201207108 cgcgctagac gatttcgatc tggacatgtt gggggacggg gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtggg gaattcgaga tgcctgtgga caggatcctg gaggcagagc ttgctgtgga acagaagagt gaccagggcg ttgagggtcc tgggggaacc gggggtagcg gcagcagccc aaatgaccct gtgactaaca tctgtcaggc agctgacaaa cagctattca cgcttgttga gtgggcgaag aggatcccac acttttcctc cttgcctctg gatgatcagg tcatattgct gcgggcaggc tggaatgaac tcctcattgc ctccttttca caccgatcca ttgatgttcg agatggcatc ctccttgcca caggtcttca cgtgcaccgc aactcagccc attcagcagg agtaggagcc atctttgatc gggtgctgac agagctagtg tccaaaatgc gtgacatgag gatggacaag acagagcttg gctgcctgag ggcaatcatt ctgtttaatc cagaggtgag gggtttgaaa tccgcccagg aagttgaact tctacgtgaa aaagtatatg ccgctttgga agaatatact agaacaacac atcccgatga accaggaaga tttgcaaaac ttttgcttcg tctgccttct ttacgttcca taggccttaa gtgtttggag catttgtttt tctttcgcct tattggagat gttccaattg atacgttcct gatggagatg cttgaatcac cttctgattc ataatctagc ctagcccccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccatigt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatctctag gcgccaccat gaagctactg tcttctatcg aacaagcatg cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgc cggaattccc ggggatccgg cctgagtgcg tagtacccga gactcagtgc gccatgaagc ggaaagagaa gaaagcacag aaggagaagg acaaactgcc tgtcagcacg acgacggtgg acgaccacat gccgcccatt atgcagtgtg aacctccacc tcctgaagca gcaaggattc acgaagtggt cccaaggttt ctctccgaca agctgttggt gacaaaccgg cagaaaaaca tcccccagtt gacagccaac cagcagttcc ttatcgccag gctcatctgg taccaggacg ggtacgagca 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 145 - 201207108 gccttctgat gaagatttga agaggattac gcagacgtgg cagcaagcgg acgatgaaaa 6060 cgaagagtcg gacactccct tccgccagat cacagagatg actatcctca cggtccaact 6120 tatcgtggag ttcgcgaagg gattgccagg gttcgccaag atctcgcagc Ctgatcaaat 61S0 tacgctgctt aaggcttgct caagtgaggt aatgatgctc cgagtcgcg gacgatacga 6240 tgcggcctca gacagtattc tgttcgcgaa caaccaagcg tacactcgcg acaa ctaccg 6300 caaggctggc atggccgagg tcatcgagga tctactgcac ttctgccggt gcatgtactc 6360 tatggcgttg gacaacatcc attacgcgct gctcacggct gtcgtcatct tttctgaccg 6420 gccagggttg gagcagccgc aactggtgga agagatccag cggtactacc tgaatacgct 6480 ccgcatctat atcctgaacc agctgagcgg gtcggcgcgt tcgtccgtca tatacggcaa 6540 gatcctctca atcctctctg agctacgcac gctcggcatg caaaactcca acatgtgcat 6600 ctccctcaag ctcaagaaca gaaagctgcc gcctttcctc gaggagatct gggatgtggc 6660 ggacatgtcg cacacccaac cgccgcctat cctcgagtcc cccacgaatc tctaggcggc 6720 ctctagagcg gccgccaccg cggggagatc cagacatgat aagatacatt gatgagtttg 6780 gacaaaccac aactagaatg cagtgaaaaa aatgcttiat ttgtgaaatt tgtgatgcta 6840 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc 6900 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct 6960 acaaatgtgg tatggctgat tatgatccgg ctgcctcgcg cgtttcggtg atgacggtga 7020 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 7080 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtc ggg gcgcagccai 7140 gaggtcgact ctagtccccg cggtggcaga tctggaaggt gctgaggtac gatgagaccc 7200 gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat taggaaccag cctgtgatgc 7260 tggatgtgac cgaggagctg aggcccgatc acttggtgct ggcctgcacc cgcgctgagt 7320 ttggctctag cgatgaagat acagattgag gtactgaaat gtgtgggcgt ggcttaaggg 7380 tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg 7440 ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc 7500 gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc 7560 ccgtcctgcc cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg 7620 agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg 7680 actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg 7740 acaagttgac ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt 7800 ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca 7860 atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt 7920 cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt 7980 cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat 8040 acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg 8100 gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt 8160 ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt 8220 taagctggga tgggtgcata cgtggggata tgagatgcat cttggactgt -146 - atttttaggt 8280 201207108

tggctatgtt cccagccata tccctccggg gattcatgtt gtgcagaacc accagcacag 8340 tgtatccggt gcacttggga aatttgtcat gtagcttaga aggaaatgcg tggaagaact 8400 tggagacgcc cttgtgacct ccaagatttt ccatgcattc gtccataatg atggcaatgg 8460 gcccacgggc ggcggcctgg gcgaagatat ttctgggatc actaacgtca tagttgtgtt 8520 ccaggatgag atcgtcatag gccattttta caaagcgcgg gcggagggtg ccagactgcg 8580 gtataatggt tccatccggc ccaggggcgt agttaccctc acagatttgc atttcccacg 8640 ctttgagttc agatgggggg atcatgtcta cctgcggggc gatgaagaaa acggtttccg 8700 gggtagggga gatcagctgg gaagaaagca ggttcctgag cagctgcgac ttaccgcagc 8760 cggtgggccc gtaaatcaca cctattaccg gctgcaactg gtagttaaga gagctgcagc 8820 tgccgtcatc cctgagcagg ggggccactt cgttaagcat gtccctgact cgcatgtttt 8880 ccctgaccaa atccgccaga aggcgctcgc cgcccagcga tagcagttct tgcaaggaag 8940 caaagttttt caacggtttg agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa 9000 gcagttccag gcggtcccac agctcggtca cctgctctac ggcatctcga tccagcatat 9060 ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc agtagtcggt gctcgtcca g 9120 acgggccagg gtcatgtctt tccacgggcg cagggtcctc gtcagcgtag tctgggtcac 9180 ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct 9240 ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt 9300 gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc agcttgccct tggaggaggc 9360 gccgcacgag gggcagtgca gacttttgag ggcgtagagc ttgggcgcga gaaataccga 9420 ttccggggag taggcatccg cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca 9480 ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt 9540 cttacctctg gtttccatga gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc 9600 cccgtataca gacttgagag gcctgtcctc gaccgatgcc cttgagagcc ttcaacccag 9660 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct 9720 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc 9780 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg 9840 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca 9900 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc g cgacgcgag 9960 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt 10020 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc 10080 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 10140 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 10200 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 10260 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 10320 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 10380 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 10440 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 10500 • 147 - 201207108 ggtatgtagg ggacagtatt gctcttgatc agattacgcg acgctcagtg tcttcaccta agtaaacttg gtctatttcg agggcttacc cagatttatc ctttatccgc cagttaatag cgtttggtat ccatgttgtg tggccgcagt catccgtaag gtatgcggcg gcagaacttt tcttaccgct catcttttac aaaagggaat attgaagcat aaaa cggtgctaca tggtatctgc cggcaaacaa cagaaaaaaa gaacgaaaac gatcct ttta gtctgacagt t tcatccata atctggcccc agcaataaac ctccatccag tttgcgcaac ggcttcattc caaaaaagcg gttatcactc atgcttttct accgagttgc aaaagtgctc gttgagatcc tttcaccagc aagggcgaca ttatcagggt gagttcttga gctctgctga accaccgctg ggatctcaag tcacgttaag aattaaaaat taccaatgct gttgcctgac agtgctgcaa cagccagccg tctattaatt gttgttgcca agctccggtt gttagctcct atggttatgg gtgactggtg tcttgcccgg atcattggaa agttcgatgt gtttctgggt cggaaatgtt tattgtctca agtggtggcc agccagttac gtagcggtgg aagatccttt ggattttggt gaagttttaa taatcagtga tccccgtcgt tgataccgcg gaagggccga gttgccggga ttgctgcagg cccaacgatc tcggtcctcc cagcactgca agtactcaac cgtcaacacg aacgttcttc aacccactcg gagcaaaaac gaatactcat tgagcggata taactacggc cttcggaaaa tttttttgtt gatcttttct catgagatta atcaatctaa ggcacctatc gtagataact agacccacgc gcgcagaagt agctagagta catcgtggtg aaggcgagtt gatcgttgtc taat tctct t caagtcat tc ggataatacc ggggcgaaaa tgcacccaac acgatacggg t aggaaggcaa actcttcctt catatttgaa tacactagaa agagttggta tgcaagcagc acggggtctg tcaaaaagga agtatatatg tcagcgatct caccggctc ggtcctgcaa agtagttcgc tcacgctcgt acatgatccc agaagtaagt actgtcatgc tgagaatagt gcgccacata ctctcaagga tgatcttcag aatgccgcaa tttcaatatt tgtatttaga 10560 10620 10680 10740 10800 10860 10920 10980 11040 11100 11160 11220 11280 11340 11400 11460 11520 11580 11640 11700 11760 11820 11824 &lt; 210 &gt; 60 &lt; 211 &gt; 11780 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; PolyDNA_from_WN-43534 &lt; 400 &gt; 60 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cct Cgagtct agagatccgg tgagtattag gcgcgcacca ggtgccgcaa laaaatatct ttattttcat tacatctgtg -148 - 60 120 180 240 300 360 420 480 540 600 660

660 C

201207108 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag ctggtgtgtg agctcatctt cctgtagatc acgcgtctcc ctcagcaagg acagcagagg accagctaag agggagagaa gcaactacag accccccctg aaaacaaccc tcagacgcca catcccctga caagctgcca ggcaggttct cttcctctca catactgacc cacggctcca ccctctctcc cctggaaagg acaccatgag cactgaaagc atgatccggg acgtggagct ggccgaggag gcgctcccca agaagacagg ggggccccag ggctccaggc ggtgcttgtt cctcagcctc ttctccttcc tgatcgtggc aggcgccacc acgctcttct gcctgctgca ctttggagtg atcggccccc agagggaaga gttccccagg gacctctctc taatcagccc tctggcccag gcagtcagat catcttctcg aaccccgagt gacaagcctg tagcccatgt tgtagcaaac cctcaagctg aggggcagct ccagtggctg aaccgccggg ccaatgccct cctggccaat ggcgtggagc tgagagataa ccagctggtg gtgccatcag agggcctgta cctcatctac tcccaggtcc tcttcaaggg ccaaggctgc ccctccaccc atgtgctcct cacccacacc atcagccgca tcgccgtctc ctaccagacc aaggtcaacc tcctctctgc catcaagagc ccctgccaga gggagacccc agagggggct gaggccaagc cctggtatga gcccatctat ctgggagggg tcttccagct ggagaagggt gaccgactca gcgctgagat caatcggccc gactatctcg actttgccga gtctgggcag gtctactttg ggatcattgc cctgtgagga ggacgaacat ccaaccttcc caaacgcctc ccctgcccca atccctttat taccccctcc ttcagacacc ctcaacctct tctggctcaa aaagagaatt gggggcttag ggtcggaacc caagcttaga actttaagca acaagaccac cacttcgaaa cctgggattc aggaatgtgt ggcctgcaca gtgaagtgct ggcaaccact aagaattcaa actggggcct ccagaactca ctggggccta cagctttgat ccctgacatc tggaatctgg agaccaggga gcctttggtt ctggccagaa tgctgcagga cttgagaaga cctcacctag aaattgacac aagtggacct taggccttcc tctctccaga tgtttccaga cttccttgag acacggagcc cagccctccc catggagcca gctccctcta tttatgtttg cacttgtgat tatttattat ttatttatta tttatttatt tacagatgaa tgtatttatt tgggagaccg gggtatcctg ggggacccaa tgtaggagct gccttggctc agacatgttt tccgtgaaaa cggagctgaa caataggctg ttcccatgta gccccctggc ctctgtgcct tcttttgatt atgtttttta aaatatttat ctgattaagt tgtctaaaca atgctgattt ggtgaccaac tgtcactcat tgctgagcct ctgctcccca ggggagttgt gtctgtaatc gccctactat tcagtggcga gaaataaagt ttgcttagaa aagaaacatg gtctccttct tggaattaat tctgcatctg cctcttcttg tgggtgggaa gaagctccct aagtcctctc tccacaggct ttaagatccc tcggacccag tcccatcctt agactcctag ggccctggag accctacata aacaaagccc 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 • 149 - 201207108 aacagaatat tccccatccc ccaggaaaca agagcctgaa cctaattacc tctccctcag 2880 ggcatgggaa tttccaactc tgggaattcc aatccttgct gggaaaatcc tgcagctcag 2940 gtgagatttc cggctgttgc agctggccag cagtccggag agagctggag Aggaggclca 3000 ttctcaggta cctgaatcac acatttaaat cggtccgcgt accgcgccgg cctccgcgcc 3060 gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg 3120 aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct 3180 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg 3240 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc 3300 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg 3360 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt 3420 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg gtacgtgcgc 3480 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt 3540 gggtcaatat gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt 3600 ttggcttttt tgttagacga gctagcgccg ccaccatggg ccctaaaaag aagcgtaaag 3660 tcgccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg 3720 cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt 3780 ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg 3840 acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtggggaat 3900 tcgagatgcc tgtggacagg atcctggagg cagagcttgc tgtggaacag aa gagtgacc 3960 agggcgttga gggtcctggg ggaaccgggg gtagcggcag cagcccaaat gaccctgtga 4020 ctaacatctg tcaggcagct gacaaacagc tattcacgct tgttgagtgg gcgaagagga 4080 tcccacactt ttcctccttg cctctggatg atcaggtcat attgctgcgg gcaggctgga 4140 atgaactcct cattgcctcc ttttcacacc gatccattga tgttcgagat ggcatcctcc 4200 ttgccacagg tcttcacgtg caccgcaact cagcccattc agcaggagta ggagccatct 4260 ttgatcgggt gctgacagag ctagtgtcca aaatgcgtga catgaggatg gacaagacag 4320 agcttggctg cctgagggca atcattctgt ttaatccaga ggtgaggggt ttgaaatccg 4380 cccaggaagt tgaacttcta cgtgaaaaag tatatgccgc tttggaagaa tatactagaa 4440 caacacatcc cgatgaacca ggaagatttg caaaactttt gcttcgtctg ccttctttac 4500 gttccatagg ccttaagtgt ttggagcatt tgtttttctt tcgccttatt ggagatgttc 4560 caattgatac gttcctgatg gagatgcttg aatcaccttc tgattcataa tctagcctag 4620 cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg 4680 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc 4740 cggaaacctg gccctgtctt ct tgacgagc attcctaggg gtct ttcccc tctcgccaaa 4800 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga 4860 caaacaacgt ctgtagcgac cctitgcagg cagcggaacc ccccacctgg cgacaggtgc 4920 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc 4980 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac 5040 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct -150 - ggggcctcgg 5100 201207108 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat ctctaggcgc caccatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcgc cgga attcccgggg atccggcctg agtgcgtagt acccgagact cagtgcgcca tgaagcggaa agagaagaaa gcacagaagg agaaggacaa actgcctgtc agcacgacga cggtggacga ccacatgccg cccattatgc agtgtgaacc tccacctcct gaagcagcaa ( "ggattcacga agtggtccca aggtttctct ccgacaagct gttggtgaca aaccggcaga aaaacatccc ccagttgaca gccaaccagc agttccttat cgccaggctc atctggtacc aggacgggta cgagcagcct tctgatgaag atttgaagag gattacgcag acgtggcagc aagcggacga tgaaaacgaa gagtcggaca ctcccttccg ccagatcaca gagatgacta tcctcacggt ccaacttatc gtggagttcg cgaagggatt gccagggttc gccaagatct cgcagcctga tcaaattacg ctgcttaagg cttgctcaag tgaggtaatg atgctccgag tcgcgcgacg atacgatgcg gcctcagaca gtattctgtt cgcgaacaac caagcgtaca ctcgcgacaa ctaccgcaag gctggcatgg ccgaggtcat cgaggatcta ctgcacttct gccggtgcat gtactctatg gcgttggaca acatccatta cgcgctgctc acggctgtcg tcatcttttc tgaccggcca gggttggagc agccgcaact ggtggaagag atccagcggt actacctgaa tacgctccgc atctatatcc tgaaccagct gagcgggtcg gcgcgttcgt ccgtcatata cggcaagatc ctctcaatcc tctctgagct acgcacgctc ggcatgcaaa actccaacat gtgcatctcc ctcaagctca agaacagaaa gctgccgcct ttcctcgagg C ;, agatctggga tgtggcggac atgtcgcaca cccaaccgcc gcctatcctc gagtccccca cgaatctcta ggcggcctct agagcggccg ccaccgcggg gagatccaga catgaiaaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgagg tcgactctag tccccgcggt ggcagatctg gaaggtgctg aggtacgatg agacccgcac caggtgcaga ccctgcgagt gtggcggtaa acatattagg aaccagcctg tgatgctgga tgtgaccgag gagctgaggc ccgatcactt ggtgctggcc tgcacccgcg ctgagtttgg ctctagcgat gaagatacag attgaggtac tgaaatgtgt 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 -151 - 201207108 gggcgtggct taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc 7380 tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc 7440 tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc 7500 agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg 7560 tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc 7620 cgcgggattg tgactgactt tgctttcctg agcccgcttg caagcagtgc agcttcccgt 7680 tcatccgccc gcgatgacaa gttgacggct cttttggcac aattggattc tttgacccgg 7740 gaacttaatg tcgtttctca gcagctgttg gatctgcgcc agcaggtttc tgccctgaag 7800 gcttcctccc ctcccaatgc ggtttaaaac ataaataaaa aaccagactc tgtttggatt 7860 tggatcaagc aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg gtaggcccgg 7920 gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg gtaaaggtga 7980 ctctggatgt tcagatacat gggcataagc ccgtctctgg ggtggaggta gcaccactgc 8040 agagcttcat gctgcggggt ggtgttgtag atgatccagt cgtagcagga Gcgctgggcg 8100 tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc cttggtgtaa 8160 gtgtttacaa agcggttaag ctgggatggg tgcatacgtg gggatatgag atgcatcttg 8220 gactgtattt ttaggttggc tccggggatt tatgttccca gccatatccc tttttacaaa gcgcgggcgg 8520 catgttgtgc 8280 agaaccacca gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag cttagaagga 8340 aatgcgtgga agaacttgga gacgcccttg tgacctccaa gattttccat gcattcgtcc 8400 ataatgatgg caatgggccc acgggcggcg gcctgggcga agatatttct gggatcacta 8460 acgtcatagt tgtgttccag gatgagatcg tcataggcca agggtgccag actgcggtat aatggttcca tccggcccag gggcgtagtt accctcacag 8580 atttgcattt cccacgcttt gagttcagat ggggggatca tgtctacctg cggggcgatg 8640 aagaaaacgg tttccggggt aggggagatc agctgggaag aaagcaggtt cctgagcagc 8700 tgcgacttac cgcagccggt gggcccgtaa atcacaccta ttaccggctg caactggtag 8760 ttaagagagc tgcagctgcc gtcatccctg agcagggggg ccacttcgtt aagcatgtcc 8820 ctgactcgca tgttttccct gaccaaatcc gccagaaggc gctcgccgcc cagcgatagc 8880 agttcttgca aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt aggcatgctt 8940 ttg agcgttt gaccaagcag ttccaggcgg tcccacagct cggtcacctg ctctacggca 9000 tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg tacggcagta 9060 gtcggtgctc gtccagacgg gccagggtca tgtctttcca cgggcgcagg gtcctcgtca 9120 gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc agggtgcgct 9180 tgaggctggt cctgctggtg ctgaagcgct gccggtcttc gccctgcgcg tcggccaggt 9240 agcatttgac catggtgtca tagtccagcc cctccgcggc gtggcccttg gcgcgcagct 9300 tgcccttgga ggaggcgccg cacgaggggc agtgcagact tttgagggcg tagagcttgg 9360 gcgcgagaaa taccgattcc ggggagtagg catccgcgcc gcaggccccg cagacggtci 9420 cgcattccac gagccaggtg agctctggcc gttcggggtc aaaaaccagg tttcccccat 9480 gctttttgat gcgtttctta cctctggttt ccatgagccg gtgtccacgc tcggtgacga 9540 aaaggctgtc cgtgtccccg tatacagact tgagaggcct gtcctcgacc -152 - gatgcccttg 9600 201207108

agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca 9660 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc 9720 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta 9780 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc 9840 ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg 9900 gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc 9960 atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga 10020 cagcttcaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 10080 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 10140 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 10200 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 10260 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 10320 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 10380 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc ca ctggtaac 10440 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 10500 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 10560 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 10620 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 10680 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 10740 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 10800 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 10860 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 10920 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 10980 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 11040 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 11100 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc 11160 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 11220 cgagttacat gatcccccat gttgtgcaaa aa agcggtta gctccttcgg tcctccgatc 11280 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 11340 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 11400 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat 11460 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 11520 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 11580 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 11640 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga Aatgttgaat actcatactc 11700 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 11760 tttgaatgta tttagaaaaa 11780 • 153 - 201207108 &lt;210> 61 &lt;211&gt; 12196 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; Ad-RTS- hIL-12 is WN2823 &lt;400&gt; 61 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 60 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 120 attaattaag ctagcatcat caataatata ccttattttg gattgaagcc aatatgataa 180 tgagggggtg gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag 240 tgtggcggaa gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa 300 aagtgacgtt tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta 360 ggcggatgtt gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa 420 actgaataag aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg 480 tctagggaga tccggtaccg gcgcgcgcgc cgtttggccg cctcgagtct agagatccgg 540 tgagtattag gcgcgcacca ggtgccgcaa taaaatatct ttattttcat tacatctgtg 600 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa 660 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct 720 ctatcgataa tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag 780 tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 840 tgtcctccga gcggagactc ttcgaaggaa gaggggcggg gtcgatcgac cccgcccctc 900 ttccttcgaa ggaagagggg cggggtcgaa gacctagagg gtatataatg ggtgccttag 960 ctggtgtgtg agctcatctt cctgtagatc acgcgtgcca ccatgggtca ccagcagttg 1020 Gtca tctctt ggttttccct ggtttttctg gcatctcccc tcgtggccat atgggaactg 1080 aagaaagatg tttatgtcgt agaattggat tggtatccgg atgcccctgg agaaatggtg 1140 gtcctcacct gtgacacccc tgaagaagat ggtatcacct ggaccttgga ccagagcagt 1200 gaggtcttag gctctggcaa aaccctgacc atccaagtca aagagtttgg agatgctggc 1260 cagtacacct gtcacaaagg aggcgaggtt ctaagccatt cgctcctgct gcttcacaaa 1320 aaggaagatg gaatttggtc cactgatatt ttaaaggacc agaaagaacc caaaaataag 1380 acctttctaa gatgcgaggc caagaattat tctggacgtt tcacctgctg gtggctgacg 1440 acaatcagta ctgatttgac attcagtgtc aaaagcagca gaggctcttc tgacccccaa 1500 ggggtgacgt gcggagctgc tacactctct gcagagagag tcagagggga caacaaggag 1560 tatgagtact cagtggagtg ccaggaggac agtgcctgcc cagctgctga ggagagtctg 1620 cccattgagg tcatggtgga tgccgttcac aagctcaagt atgaaaacta caccagcagc 1680 ttcttcatca gggacatcat caaacctgac ccacccaaga acttgcagct gaagccatta 1740 aagaattctc ggcaggtgga ggtcagctgg gagtaccctg acacctggag tactccacai 1800 tcctacttct ccctgacatt ctgcgttcag gtccagggca agagcaagag agaaaagaaa 1860 gatagagtct Tcacggacaa gacctcagcc acggtcatct gccgcaaaaa tgccagcatt 1920 agcgtgcggg cccaggaccg ctactatagc tcatcttgga gcgaatgggc atctgtgccc 1980 -154 -

201207108 tgcagttagg ttgggcgagc tcgaattcat tgatcccccg ggctgcagga attcgatatc aagctcggga tccgaattcc gccccccccc cccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat gggtccagcg cgcagcctcc tccttgtggc taccctggtc ctcctggacc acctcagttt ggccagaaac ctccccgtgg ccactccaga cccaggaatg ttcccatgcc ttcaccactc ccaaaacctg ctgagggccg tcagcaacat gctccagaag gccagacaaa ctctagaatt ttacccttgc acttctgaag agattgatca tgaagatatc acaaaagata aaaccagcac agtggaggcc tgtttaccat tggaattaac caagaatgag agttgcctaa attccagaga gacctctttc ataactaatg ggagttgcct ggcctccaga aagacctctt ttatgatggc cctgtgcctt agtagtattt atgaagactt gaagatgtac caggtggagt tcaagaccat gaatgcaaag cttctgatgg atcctaagag gcagatcttt ctagatcaaa acatgctggc agttattgat gagctgatgc aggccctgaa tttcaacagt gagactgtgc cacaaaaatc ctcccttgaa gaaccggatt tttataaaac taaaatcaag ctctgcatac ttcttcatgc tttcagaatt cgggcagtga ctattgatag agtgatgagc tatctgaatg cttcctaacg tacgtcgaca tcgagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgggcg cgccggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctgggtac gtgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgagcta gcgccgccac catgggccct aaaaagaagc gtaaagtcgc ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 • 155 · 201207108 gggtccggga tttacccccc acgactccgc cccctacggc gctctggata tggccgactt 4260 cgagtttgag cagatgttta ccgatgccct tggaattgac gagtacggtg gggaattcga 4320 gatgcctgtg gacaggatcc tggaggcaga gcttgctgtg Gaacagaaga gtgaccaggg 4380 cgttgagggt cctgggggaa ccgggggtag cggcagcagc ccaaatgacc ctgtgactaa 4440 catctgtcag gcagctgaca aacagctatt cacgcttgtt gagtgggcga agaggatccc 4500 acacttttcc tccttgcctc tggatgatca ggtcatattg ctgcgggcag gctggaatga 4560 actcctcatt gcctcctttt cacaccgatc cattgatgtt cgagatggca tcctccttgc 4620 cacaggtctt cacgtgcacc gcaactcagc ccattcagca ggagtaggag ccatctttga 4680 tcgggtgctg acagagctag tgtccaaaat gcgtgacatg aggatggaca agacagagct 4740 tggctgcctg agggcaatca ttctgtttaa tccagaggtg aggggtttga aatccgccca 4800 ggaagttgaa cttctacgtg aaaaagtata tgccgctttg gaagaatata ctagaacaac 4860 acatcccgat gaaccaggaa gatttgcaaa acttttgctt cgtctgcctt ctttacgttc 4920 cataggcctt aagtgtttgg agcatttgtt tttctttcgc cttattggag atgttccaat 4980 tgatacgttc ctgatggaga tgcttgaatc accttctgat tcataatcta gcctagcccc 5040 cctctccctc ccccccccct aacgt tactg gccgaagccg cttggaataa ggccggtgtg 5100 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga 5160 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa 5220 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa 5280 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc a cctggcgac aggtgcctct 5340 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg 5400 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg 5460 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca 5520 catgctttac atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga 5580 cgtggttttc ctttgaaaaa cacgatctct aggcgccacc atgaagctac tgtcttctat 5640 cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa 5700 gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc 5760 tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct 5820 atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca 5880 ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt 5940 cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat 6000 aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc 6060 gccggaattc ccggggatcc ggcctgagtg cgtagtaccc gagactcagt gcgccatgaa 6120 gcggaaagag aagaaagcac agaaggagaa ggac aaactg cctgtcagca cgacgacggt 6180 ggacgaccac atgccgccca ttatgcagtg tgaacctcca cctcctgaag cagcaaggat 6240 tcacgaagtg gtcccaaggt ttctctccga caagctgttg gtgacaaacc ggcagaaaaa 6300 catcccccag ttgacagcca accagcagtt cct tatcgcc aggctcatct ggtaccagga 6360 cgggtacgag cagccttctg atgaagattt gaagaggatt acgcagacgt ggcagcaagc 6420 ggacgatgaa aacgaagagt cggacactcc cttccgccag atcacagaga -156 - tgactatcct 6480 201207108

cacggtccaa cttatcgtgg agttcgcgaa gggattgcca gggttcgcca agatctcgca 6540 gcctgatcaa attacgctgc ttaaggcttg ctcaagtgag gtaatgatgc tccgagtcgc 6600 gcgacgatac gatgcggcct cagacagtat tctgttcgcg aacaaccaag cgtacactcg 6660 cgacaactac cgcaaggctg gcatggccga ggtcatcgag gatctactgc acttctgccg 6720 gtgcatgtac tctatggcgt tggacaacat ccattacgcg ctgctcacgg ctgtcgtcat 6780 cttttctgac cggccagggt tggagcagcc gcaactggtg gaagagatcc agcggtacta 6840 cctgaatacg ctccgcatct atatcctgaa ccagctgagc gggtcggcgc gttcgtccgt 6900 catatacggc aagatcctct caatcctctc tgagctacgc acgctcggca tgcaaaactc 6960 caacatgtgc atctccctca agctcaagaa cagaaagctg ccgcctttcc tcgaggagat 7020 ctgggatgtg gcggacatgt cgcacaccca accgccgcct atcctcgagt cccccacgaa 7080 tctctaggcg gcctctagag cggccgccac cgcggggaga tccagacatg ataagataca 7140 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 7200 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca 7260 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagc a 7320 agtaaaacct ctacaaatgt ggtatggctg attatgatcc ggctgcctcg cgcgtttcgg 7380 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 7440 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 7500 gggcgcagcc atgaggtcga ctctagtccc cgcggtggca gatctggaag gtgctgaggt 7560 acgatgagac ccgcaccagg tgcagaccct gcgagtgtgg cggtaaacat attaggaacc 7620 agcctgtgat gctggatgtg accgaggagc tgaggcccga tcacttggtg ctggcctgca 7680 cccgcgctga gtttggctct agcgatgaag atacagattg aggtactgaa atgtgtgggc 7740 gtggcttaag ggtgggaaag aatatataag gtgggggtct tatgtagttt tgtatctgtt 7800 ttgcagcagc cgccgccgcc atgagcacca actcgtttga tggaagcatt gtgagctcat 7860 atttgacaac gcgcatgccc ccatgggccg gggtgcgtca gaatgtgatg ggctccagca 7920 ttgatggtcg ccccgtcctg cccgcaaact ctactacctt gacctacgag accgtgtctg 7980 gaacgccgtt ggagactgca gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg 8040 ggattgtgac tgactttgct ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat 8100 ccgcccgcga tgacaagttg acggctcttt tggcacaatt ggattctttg a cccgggaac 8160 ttaatgtcgt ttctcagcag ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt 8220 cctcccctcc caatgcggtt taaaacataa ataaaaaacc agactctgtt tggatttgga 8280 tcaagcaagt gtcttgctgt ctttatttag gggttttgcg cgcgcggtag gcccgggacc 8340 agcggtctcg gtcgttgagg gtcctgtgta ttttttccag gacgtggtaa aggtgactct 8400 ggatgttcag atacatgggc ataagcccgt ctctggggtg gaggtagcac cactgcagag 8460 cttcatgctg cggggtggtg ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt 8520 gcctaaaaat gtctttcagt agcaagctga ttgccagggg caggcccttg gtgtaagtgt 8580 ttacaaagcg gttaagctgg gatgggtgca tacgtgggga tatgagatgc atcttggact 8640 gtatttttag gttggctatg ttcccagcca tatccctccg gggattcatg ttgtgcagaa 8700 • 157 - 201207108 ccaccagcac agtgtatccg gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg 8760 cgtggaagaa cttggagacg cccttgtgac ctccaagatt ttccatgcat tcgtccataa 8820 tgatggcaat gggcccacgg gcggcggcct gggcgaagat atttctggga tcactaacgt 8880 catagttgtg ttccaggatg agatcgtcat aggccatttt tacaaagcgc gggcggaggg 8940 tgccagactg cggtataatg gttccat ccg gcccaggggc gtagttaccc tcacagattt 9000 gcatttccca cgctttgagt tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga 9060 aaacggtttc cggggtaggg gagatcagct gggaagaaag caggttcctg agcagctgcg 9120 acttaccgca gccggtgggc ccgtaaatca cacctattac cggctgcaac tggtagttaa 9180 gagagctgca gctgccgtca tccctgagca ggggggccac ttcgttaagc atgtccctga 9240 ctcgcatgtt ttccctgacc aaatccgcca gaaggcgctc gccgcccagc gatagcagtt 9300 cttgcaagga agcaaagttt ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga 9360 gcgtttgacc aagcagttcc aggcggtccc acagctcggt cacctgctct acggcatctc 9420 gatccagcat atctcctcgt ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg 9480 gtgctcgtcc agacgggcca gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt 9540 agtctgggtc acggtgaagg ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag 9600 gctggtcctg ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca 9660 tttgaccatg gtgtcatagt ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc 9720 cttggaggag gcgccgcacg aggggcagtg cagacttttg agggcgtaga gcttgggcgc 9780 gagaaatacc gattccgggg agtaggcatc cgcgccgcag gccccgcaga cggtctcgca 9840 ttccacgagc caggtgagct ctggccgttc ggggtcaaaa accaggtttc ccccatgctt 9900 tttgatgcgt t tct tacctc tggtttccat gagccggtgt ccacgctcgg tgacgaaaag 9960 gctgtccgtg tccccgtata cagacttgag aggcctgtcc tcgaccgatg cccttgagag 10020 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta 10080 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt 10140 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg 10200 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg 10260 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt 10320 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg 10380 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc 10440 ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 10500 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 10560 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 10620 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 10680 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 10740 gctgtgtgca cgsscccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 10800 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 10860 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 10920 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt -158 - accttcggaa 10980

C

201207108 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttitg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaa &lt; 210 &gt; 62 &lt; 211 &gt; 12166 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; Ad-RTS-mIL-12 are WN2539 &lt; 400 &gt; 62 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaaaattaa ttaagctagc atcatcaata atatacctta ttttggattg aagccaatat gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg Cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactca t agcgcgtaat atttgtctag ggagatccgg taccggcgcg cgcgccgttt ggccgcctcg agtctagaga tccggtgagt gattaggcgc gcaccaggtg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa 11040 11100 11160 11220 11280 11340 11400 11460 11520 11580 11640 11700 11760 11820 11880 11940 12000 12060 12120 12180 12196 60 120 180 240 300 360 420 480 540 600 159 - 660 201207108 aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac 720 atttctctat cgataatgca ggtcggagta ctgtcctccg agcggagtac tgtcctccga 780 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg 840 gagtactgtc ctccgagcgg agactcttcg aaggaagagg ggcggggtcg atcgaccccg 900 cccctcttcc ttcgaaggaa gaggggcggg gtcgaagacc tagagggtat ataatgggtg 960 ccttagctgg tgtgtgagct catcttcctg tagatcacgc gtgccaccat gtgtcctcag 1020 aagctaacca tctcctggtt tgccatcgtt Ttgctggtgt ciccactcat ggccatgtgg 1080 gagctggaga aagacgttta tgttgtagag gtggactgga ctcccgatgc ccctggagaa 1140 acagtgaacc tcacctgtga cacgcctgaa gaagatgaca tcacciggac ctcagaccag 1200 agacatggag tcataggctc tggaaagacc ctgaccatca ctgtcaaaga gtttctagat 1260 gctggccagt acacctgcca caaaggaggc gagactctga gccactcaca tctgctgctc 1320 cacaagaagg aaaatggaat ttggtccact gaaattttaa aaaatttcaa aaacaagact 1380 ttcctgaagt gtgaagcacc aaattactcc ggacggttca cgtgctcatg gctggtgcaa 1440 agaaacatgg acttgaagtt caacatcaag agcagtagca gtccccccga ctctcgggca 1500 gtgacatgtg gaatggcgtc tctgtctgca gagaaggtca cactggacca aagggactat 1560 gagaagtatt cagtgtcctg ccaggaggat gtcacctgcc caactgccga ggagaccctg 1620 cccattgaac tggcgttgga agcacggcag cagaataaat atgagaacta cagcaccagc 1680 ttcttcatca gggacatcat caaaccagac ccgcccaaga acttgcagat gaagcctttg 1740 aagaactcac aggtggaggt cagctgggag taccctgact cctggagcac tccccattcc 1800 tacttctccc tcaagttctt tgttcgaatc cagcgcaaga aagaaaagat gaaggagaca 1860 gaggaggggt gtaaccagaa aggtgcgttc ctcgtagaga agacatctac cgaagtccaa 1920 tgcaaaggcg ggaatgtctg cgtgcaagct caggatcgct attacaattc ctcatgcagc 1980 aagtgggcat gtgttccctg cag ggtccga tcctaggatg caacggatcc gaattccgcc 2040 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 2100 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg 2160 gaaacctggc cctgtcttct tgacgagcat ctttcccctc tcgccaaagg 2220 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt ct tgaagaca 2280 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 2340 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 2400 cgttgtgagt tggatagttg tcctaggggt tggaaagagt caaatggctc tcctcaagcg tattcaacaa 2460 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg 2520 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg 2580 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg ggtcaatcac 2640 gctacctcct ctttttggcc acccttgccc tcctaaacca cctcagtttg gccagggtca 2700 11CC2.gt C t C tggacctgcc aggtgtctta gccagtcccg aaacctgctg aagaccacag 2760 atgacatggt gaagacggcc agagaaaagc tgaaacatta ttcctgcact Gctgaagaca 2820 tcgatcatga Agacatcaca cgggaccaaa ccagcacatt gaagacctgt t taccactgg 2880 aactacacaa gaacgagagt tgcctggcta ctagagagac ttcttccaca -160 - acaagaggga 2940 201207108

gctgcctgcc cccacagaag acgtctttga tgatgaccct gtgccttggt agcatctatg 3000 aggacttgaa gatgtaccag acagagttcc aggccatcaa cgcagcactt cagaatcaca 3060 accatcagca gatcattcta gacaagggca tgctggtggc catcgatgag ctgatgcagt 3120 ctctgaatca taatggcgag actctgcgcc agaaacctcc tgtgggagaa gcagaccctt 3180 acagagtgaa aatgaagctc tgcatcctgc ttcacgcctt cagcacccgc gtcgtgacca 3240 tcaacagggt gatgggctat ctgagctccg cctgagtcga catcgagaac ttgtttattg 3300 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 3360 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 3420 gcgccggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag 3480 cgcigccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg 3540 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt 3600 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc 3660 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc 3720 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggatt t 3780 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg 3840 ggctgggtac gtgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt 3900 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc 3960 taaattctgg ccgtttttgg CtUtttgtt agacgagcta gcgccgccac catgggccct 4020 aaaaagaagc gtaaagtcgc ccccccgacc gatgtcagcc tgggggacga gctccactta 4080 gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg 4140 ttgggggacg gggattcccc ggggccggga tttacccccc acgactccgc cccctacggc 4200 gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac 4260 gagtacggtg gggaattcga gatgcctgtg gacaggatcc tggaggcaga gcttgctgtg 4320 gaacagaaga gtgaccaggg cgttgagggt cctgggggaa ccgggggtag cggcagcagc 4380 ccaaatgacc ctgtgactaa catctgtcag gcagctgaca aacagctatt cacgcttgtt 4440 gagtgggcga agaggatccc acacttttcc tccttgcctc tggatgatca ggtcatattg 4500 ctgcgggcag gctggaatga actcctcatt gcctcctttt cacaccgatc cattgatgtt 4560 cgagatggca tcctccttgc cacaggtctt cacgtgcacc gcaactcagc cc attcagca 4620 ggagtaggag ccatctttga tcgggtgctg acagagctag tgtccaaaat gcgtgacatg 4680 aggatggaca agacagagct tggctgcctg agggcaatca ttctgtttaa tccagaggtg 4740 aggggtttga aatccgccca ggaagttgaa cttctacgtg aaaaagtata tgccgctttg 4800 gaagaatata ctagaacaac acatcccgat gaaccaggaa gatttgcaaa acttttgctt 4860 cgictgcctt ctttacgttc cataggcctt aagtgtttgg agcatttgtt tttctttcgc 4920 cttattggag atgttccaat tgatacgttc ctgatggaga tgcttgaatc accttctgat 4980 tcataatcta gcggccctag cccccctctc cctccccccc ccctaacgtt actggccgaa 5040 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt 5100 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 5160 -161 - 201207108 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 5220 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 5280 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 5340 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 5400 tctcctcaag cgtattcaac aaggggctgaaggatgccca gaaggtaccc cattgtatgg 5460 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg 5520 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat ctctagggcc 5580 gcgccaccat gaagctactg tcttctatcg aacaagcatg cgatatttgc cgacttaaaa 5640 agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac aactgggagt 5700 gtcgctactc tcccaaaacc aaaaggtctc cgctgactag ggcacatctg acagaagtgg 5760 aatcaaggct agaaagactg gaacagctat ttctactgat ttttcctcga gaagaccttg 5820 acatgatttt gaaaatggat tctttacagg atataaaagc attgttaaca ggattatttg 5880 tacaagataa tgtgaataaa gatgccgtca cagatagatt ggcttcagtg gagactgata 5940 tgcctctaac attgagacag catagaataa gtgcgacatc atcatcggaa gagagtagta 6000 acaaaggtca aagacagttg actgtatcgc cggaattccc ggggatccgg cctgagtgcg 6060 tagtacccga gactcagtgc gccatgaagc ggaaagagaa gaaagcacag aaggagaagg 6120 acaaactgcc tgtcagcacg acgacggtgg acgaccacat gccgcccatt atgcagtgtg 6180 aacctccacc tcctgaagca gcaaggattc acgaagtggt cccaaggttt ctctccgaca 6240 agctgttgga gacaaaccgg cag aaaaaca tcccccagtt gacagccaac cagcagttcc 6300 ttatcgccag gctcatctgg taccaggacg ggtacgagca gccttctgat gaagatttga 6360 agaggattac gcagacgtgg cagcaagcgg acgatgaaaa cgaagagtcg gacactccct 6420 tccgccagat cacagagatg actatcctca cggtccaact tatcgtggag ttcgcgaagg 6480 gattgccagg gttcgccaag atctcgcagc ctgatcaaat tacgctgctt aaggcttgct 6540 caagtgaggt aatgatgctc cgagtcgcgc gacgatacga tgcggcctca gacagtattc 6600 tgttcgcgaa caaccaagcg tacactcgcg acaactaccg caaggctggc atggccgagg 6660 tcatcgagga tctactgcac ttctgccggt gcatgtactc tatggcgttg gacaacatcc 6720 attacgcgct gctcacggct gtcgtcatct tttctgaccg gccagggttg gagcagccgc 6780 aactggtgga agagatccag cggtactacc tgaatacgct ccgcatctat atcctgaacc 6840 agctgagcgg gtcggcgcgt tcgtccgtca tatacggcaa gatcctctca atcctctctg 6900 agctacgcac gctcggcatg caaaactcca acatgtgcat ctccctcaag ctcaagaaca 6960 gaaagctgcc gcctttcctc gaggagatct gggatgtggc ggacatgtcg cacacccaac 7020 cgccgcctat cctcgagtcc cccacgaatc tctaggcggc ctctagagcg gccgccaccg 7080 cggggagatc cagaca tgat aagatacatt gatgagtttg gacaaaccac aactagaatg 7140 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt 7200 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag 7260 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat 7320 tatgatcccg gctgcctcgc gcgtttcggt gatgacggtg aaaacctctt gacacatgca 7380 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac •] 62 - aagcccgtca 7440

201207108 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgaggtcga ctctagtccc cgcggtggca gatctggaag gtgctgaggt acgatgagac ccgcaccagg tgcagaccct gcgagtgtgg cggtaaacat attaggaacc agcctgtgat gctggatgtg accgaggagc tgaggcccga tcacttggtg ctggcctgca cccgcgctga gtttggctct agcgatgaag atacagattg aggtactgaa atgtgtgggc gtggcttaag ggtgggaaag aatatataag gtgggggtct tatgtagttt tgtatctgtt ttgcagcagc cgccgccgcc atgagcacca actcgtttga tggaagcatt gtgagctcat atttgacaac gcgcatgccc ccatgggccg gggtgcgtca gaatgtgatg ggctccagca ttgatggtcg ccccgtcctg cccgcaaact ctactacctt gacctacgag accgtgtctg gaacgccgtt ggagactgca gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac tgactttgct ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga tgacaagttg acggctcttt tggcacaatt ggattctttg acccgggaac ttaatgtcgt ttctcagcag ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt taaaacataa ataaaaaacc agactctgtt tggatttgga tcaagcaagt gtcttgctgt ctttatttag gggttttgcg cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg gtcctgtgta ttttttccag gacgtggtaa aggtgactct ggatgttcag atacatgggc ataagcccgt ctctggggtg gaggtagcac cactgcagag cttcatgctg cggggtggtg ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt agcaagctga ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg gatgggtgca tacgtgggga tatgagatgc atcttggact gtatttttag gttggctatg ttcccagcca tatccctccg gggattcatg ttgtgcagaa ccaccagcac agtgtatccg gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa cttggagacg cccttgtgac ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg gcggcggcct gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg agatcgtcat aggccatttt tacaaagcgc gggcggaggg tgccagactg cggtataatg gttccatccg gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtaggg gagatcagct gggaagaaag caggttcctg agcagctgcg acttaccgca gccggtgggc ccgtaaatca cacctattac cgggtgcaac tggtagttaa gagagctgca gctgccgtca tccctgagca ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc aaatccgcca gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc aggcggtccc acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt ccagcccctc 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9180 9240 9300 9360 9420 9480 9540 9600 9660 163 - 201207108 cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcacg aggggcagtg 9720 cagacttttg agggcgtaga gcttgggcgc gagaaatacc gattccgggg agtaggcatc 9780 cgcgccgcag gccccgcaga cggtctcgca t tccacgagc Caggtgagct ctggccgttc 9840 ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat 9900 gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cag acttgag 9960 aggcctgtcc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg 10020 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag 10080 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga 10140 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg 10200 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg 10260 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca 10320 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca 10380 ggcaggtaga tgacgaccat cagggacagc t tcaaggcca gcaaaaggcc aggaaccgta 10440 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 10500 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 10560 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 10620 ccgcctttct cccttcggga agcgtggcgc 11 tctcatag ctcacgctgt aggtatctca 10680 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 10740 accgctgcgc ct tatccggt aactatcgtc t tgagtccaa cccggtaaga cacgacttat 10800 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 10860 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 10920 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 10980 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 11040 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 11100 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 11160 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 11220 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 11280 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 11340 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 11400 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 11460 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 11520 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 11580 tcagctccgg t tcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 11640 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 11700 tcatggttat ggcagcactg cataattctc tiactgtcat gccatccgta agatgctttt 11760 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagt t 11820 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc] 1880 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg -164 - ctgttgagat 11940 201207108 ccagttcgat gtaacccact cgtgcaccca Actgatcttc agcatctttt actttcacca 12000 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 12060 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 12120 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaa 12166

)

Claims (1)

201207108 VII. Application for Patent Park: 1 · A carrier for egg yolk that conditionally exhibits the function of one or more immunomodulators, comprising a polynucleotide encoding a gene switch, wherein the polynucleotide comprises (1) at least a transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor and (2) a polynucleotide encoding one or more proteins having an immunomodulatory function, It is operably linked to a promoter that is activated by the ligand-dependent transcription factor. 2) The carrier of claim 3, wherein the one or more immunomodulators are selected from the group consisting of IL-1, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8 , IL-9, IL10R DN or subunits, 11^15, 11^-18, 11^-21, IL-23, IL-24, IL-27, GM-CSF, IFN-α, IFN-γ , IFN-a 1 , IFN a 2 , IL -1 5 - R - a , CCL 3 ( ΜIP -1 a), CCL5 (RANTES), CCL7 (MCP3), XCL1 (lymphocyte chemotactic factor), CXCL1 (MGSA -α) , CCR7 , CCL19 ( MIP-3b ) , ( I CXCL9 ( MIG ) , CXCL 1 0 (IP-1 0) , CXCL 1 2 (SDF-1), CCL21 (6Ckine), OX40L, 4-1BBL &gt ; CD40, CD70, GITRL, LIGHT, b-defensin, HMGB1, Flt3L, IFN-β, TNF-α, dnFADD, BCG, TGF-a, PD-L 1 RNAi, PD-L 1 antisense oligonucleotide , TGFbRII DN, ICOS-L, S100, CD40L, p53, survivin, p53-survivin fusion, MAGE3, PSA or PSMA. The carrier of claim 1 or 2, wherein the vector is selected from the group consisting of a plastid, an adenovirus, a retrovirus, an adeno-associated virus, a prion, a baculovirus, a vaccinia virus, a herpes simplex virus, an EB ( Epstein- 201207108 Barr) Virus, adenovirus, geminivirus, cauliflower mosaic virus, liposome, charged lipid (cell transfectin), DNA_protein complex or biopolymer. 4.  The vector of any one of claims 1 to 3, wherein the vector is an adenoviral vector. 5.  The vector of any one of claims 1 to 4, which further comprises a polynucleotide encoding a protein having the function of IL-12. 6.  The vector of claim 5, wherein the polynucleotide encoding the protein or the protein having the function of an immunomodulator and the polynucleotide encoding the protein having the function of IL-1 2 are in the gene The switch is under the control of a regulated promoter. 7 . The vector of any one of claims 1 to 6, wherein the gene is associated with a gene switch based on the ecdysone receptor (EcR). 8 . The vector of any one of claims 1 to 7, wherein the polynucleotide encoding the gene switch comprises a first transcription factor sequence under the control of the first promoter and a second under the control of the second promoter A transcription factor sequence in which proteins encoded by the first transcription factor sequence and the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. 9. The vector of any one of claims 1 to 7 wherein the polynucleotide encoding the gene switch comprises a first transcription factor sequence and a second transcription factor sequence under the control of a promoter, wherein the first transcription is The factor sequence and the protein encoded by the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. -2- 201207108 10. The vector of claim 9, wherein the first transcription factor sequence and the second transcription factor sequence are linked by an EMCV internal ribosome entry site (IRES). Hey. The vector of any one of claims 1 to 10, wherein the polynucleotide encoding one or more proteins having the function of an immunomodulator encodes a human immunomodulator. 1 2 . The vector of any one of claims 5 to 11, wherein the polynucleotide encoding the protein having the function of IL-12 encodes human IL-12 〇 1 3 . The vector of any one of claims 1 to 12, wherein the immunomodulator is TNF-α. 14. The vector of claim 13, wherein the immunomodulator is human TNF-α. 1 5 . The vector of any one of claims 1 to 4, wherein the immunomodulator comprises at least 80%, 85% &gt; 90%, 95 with SEQ ID NO: 37 (human TNF-a). %, 9 6% '97%, 9 8%, 99% or 100% - the amino acid sequence. 1 6 . The vector of any one of claims 1 to 15 wherein the polynucleotide encoding one or more proteins having the function of an immunomodulator is codon-optimized. 17.  The vector of claim 16, wherein the polynucleotide encoding one or more proteins having the function of an immunomodulator further comprises a nucleic acid sequence encoding a signal peptide. 18.  The vector of claim 17 wherein the nucleic acid sequence encoding the signal -3-201207108 peptide is codon-optimized. 19.  The vector of claim 18, wherein the signal peptide is selected from the group consisting of TNFOptUV or IL-2optUV. 20.  The vector of any one of claims 17 to 19 wherein the signal peptide induces enhanced TNF-a protein secretion compared to the peptide sequence encoded by the wild-type TNF-α signal peptide gene. twenty one . The carrier of any one of claims 1 to 20, which additionally includes a 5' non-translated region (UTR). twenty two. The vector of claim 21, wherein the 5' UTR is derived from TNF wild type or 5U2. 23. The vector of claim 21, wherein the 5' UTR induces an increase in the amount of mRNA encoding TNF-a, an increase in the amount of TNF-a protein expression, or both. twenty four.  The carrier of any one of claims 1 to 23, further comprising a 3' regulatory region. 25.  The vector of claim 24, wherein the 3' regulatory region is derived from a polyadenylation signal of SV40e or human growth hormone (hGH). 26.  The vector of claim 24 or 25, wherein the 3' regulatory region induces an increase in the amount of mRNA encoding TNF-a, an increase in the amount of TNF-a protein expression, or both. 27.  For example, the carrier of claim 26, which contains 5U2. UTR, codon-optimized IL-2 signal peptide (IL-2optUV) 'TNF-a encoded by a codon-optimized nucleic acid sequence and 3' regulatory region derived from hGH 201207108 28 · as claimed The vector of any one of items 1 to 27, which comprises a vector selected from the group consisting of a vector 43318, a vector 43319, a vector 43 320, a vector 43321, a vector 433 22, a vector 43 322, a vector 43323, a carrier 433 24, a carrier 43325, a carrier 43326, The nucleic acid sequence of vector 43327 or vector 43329. 29. The vector of any one of claims 1 to 28, wherein the vector for conditionally expressing the immunomodulator is directly injected into or near the tumor. 3 0. - a method of producing an immune cell or a group of therapeutic support cells (TSC), the immune cell or TSC group exhibiting a protein having the function of one or more immunomodulators, the method comprising use as in claims 1 to 29 of the scope of the patent application Any one of the vectors modifies the immune cells. 3 1 . The method of claim 30, wherein the cells are human dendritic cells. 3 2 . The method of claim 31, wherein the dendritic cells are bone marrow dendritic cells. 33. An immunocyte or TSC population of a protein exhibiting the function of one or more immunomodulators, comprising the vector of any one of claims 1-3 to 3 of the patent application. 3 4 . For example, the immune cells or the τ S C population of claim 3, wherein the cells are human dendritic cells. 35.  For example, the cell population of claim 34, wherein the dendritic cells are bone marrow dendritic cells. 36.  An in vitro engineered immune cell or TSC comprising the vector of any one of claims 1 to 30. -5- 201207108 37. For example, in vitro engineered cells or TSCs of claim 36, wherein the immune cells or TSCs are human dendritic cells. 3 8 . For example, in vitro engineered cells or TSCs of claim 3, wherein the dendritic cells are bone marrow dendritic cells. 39. A pharmaceutical composition comprising a carrier according to any one of the scope of the patent application, such as an immune cell or a TSC group in claims 3 to 35, as claimed in claims 36 to 38 An in vitro engineered immune cell or TSC or a component thereof, comprising two or more carriers, as claimed in any one of claims 30 to 30, or two or more The immune cell or TSC group of any one of the patents of claim 5, the two or more of the in vitro engineered immune TSCs of any one of claims 36 to 38, or any combination thereof. 41. A pharmaceutically acceptable carrier, such as the composition of claim 39 or 40. 42. A group according to any one of the claims of the invention, wherein the composition is suitable for oral, intravitreal, intratumoral, or intraperitoneal administration. 43. The group of any one of claims 3 to 42 wherein the cell population comprises at least 1 to 4 cells. 44. The group of any one of claims 39 to 43 wherein the cell population comprises at least 1 to 7 cells. 45. For example, any one of the immunizations of any one of the groups of claims 39 to 44, g1 to 30, any combination of dad, the first 33rd to the patented cells or containing the medical substance, intramembranous or Adult, adult, adult, -6 - 201207108 When the composition and the activating ligand are administered to a mammal in need thereof simultaneously or in any order, the composition reduces the tumor size. 46.  The composition of any one of claims 39 to 45, which prevents tumor formation when the composition and the activating ligand are administered to a mammal in need thereof simultaneously or in any order. 47.  The composition of any one of claims 39 to 46, wherein the composition treats the tumor when the composition and the activating ligand are administered simultaneously or in any order to the mammal in need thereof. 48.  (1) A carrier according to any one of claims 1 to 30, a cell population according to any one of claims 3 to 3, wherein the patent scope is 3-6 to 3 8 The in vitro engineered immune cell or TSC of any one of the items, or the composition of any one of claims 39 to 47, and (2) a therapeutically effective amount of one or more activating ligands for manufacture Use of a medicament for treating a tumor in a mammal. 49.  (1) A carrier Q according to any one of claims 3 to 3, a cell population according to any one of claims 3 to 3, and a patent scope 3-6 to 3 The in vitro engineered immune cell or TSC of any of the eight items, or the pharmaceutical composition according to any one of claims 39 to 47, and (2) a therapeutically effective amount of one or more activating ligands The use of a medicament for reducing the size of a tumor for a mammal in need thereof. A carrier group according to any one of claims 1 to 3, or a cell population according to any one of claims 3 to 3, as claimed in claim 3 An in vitro engineered immune cell or TSC of any one of items 6 to 38, or a pharmaceutical 201207108 composition according to any one of claims 30 to 47 and (2) a therapeutically effective amount Use of one or more activating ligands for the manufacture of a medicament for the prevention of tumorigenesis in a mammal in need thereof. 5 1. (1) A carrier according to any one of claims 1 to 30, a cell population according to any one of claims 3 to 3, wherein the patent scope is 3-6 to 3 8 The in vitro engineered immune cell or TSC of any one of the items, or the pharmaceutical composition according to any one of claims 39 to 47, and (2) a therapeutically effective amount of one or more activating ligands Use of a medicament for reducing, eliminating or controlling the systemic toxicity of TNF-[alpha] in a mammal in need thereof, wherein the system is administered simultaneously with, before or after administration of the carrier or composition and wherein the activating ligand The administration reduces, eliminates or controls systemic toxicity in a timely manner. 52. The use of any one of claims 48 to 51, wherein when the mammal has an adverse reaction, the administration of the activating ligand is stopped 〇5 3 as in the patent application range 48-8 to 2 The use of any of the embodiments, wherein the system is administered within one hour before or after administration of the carrier or composition. 54. The use of any one of claims 48 to 52, wherein the system is administered within 24 hours after administration of the carrier or composition. The use of any one of the claims 4 to 5, wherein the system is administered within 48 hours after administration of the carrier or composition. 5 6 . The use of any one of claims 4 to 5, wherein the vector is not contained in a cell and is administered intratumorally to the tumor environment of the mammal. -8- 201207108 5 7 . The use of claim 56, wherein the immune cell or TSC is not administered intratumorally with the vector. 5 8. The use of any one of claims 48 to 57, wherein the tumor is a benign tumor. 59. The use of any one of claims 48 to 57, wherein the tumor is a malignant tumor. 6 0. The use of any one of claims 48 to 57, wherein the tumor is melanoma. 6 1 . The use of any one of claims 4 to 75, wherein the tumor is malignant melanoma skin cancer. 62 . The use of any one of the claims 4 to 61, wherein the system is dioxin. 63.  The use of any one of claims 48 to 62, wherein the system is selected from the group consisting of RG-1 15819, RG-115932 or RG-115830. 64.  The use of any one of claims 48 to 61, wherein the system is amidoxime or oxadiazoline.  The use of any one of claims 48 to 64, wherein the system is administered within one hour before or after administration of the vector, immune cell or TSC population, in vitro engineered immune cells or composition Cast. 66.  The use according to any one of claims 48 to 64, wherein the system is administered within 24 hours after administration of the vector, immune cell or TSC population, in vitro engineered immune cells or composition. Give. 67.  The use according to any one of the claims 48 to 64, wherein the system is administered after the carrier, the immune cell or the TSC group, the immune cell or composition after the in vitro work -9-201207108 It is administered within 8 hours. 6 8 .  An immunological cell or a TSC group according to any one of claims 1 to 30 of the patent application, as in any one of claims 33 to 35, as claimed in claim 36-38 A method for the therapeutic effect of a therapeutic composition of a subcutaneously engineered immune cell, or a composition according to any one of claims 39 to 47, for a patient, comprising: Measuring the amount of activity or activity or both of interferon gamma (ιι?Ν-γ) in the first biological sample, the first biological sample being administered to the vector, the immune cell or the TSC group, and in vitro The engineered immune cells or composition are obtained from the patient to be measured before, thereby generating a control amount; (b) administering to the patient to be measured, as in any one of claims 1 to 30 of the patent application. The vector, the immune cell or the TSC group according to any one of claims 3 to 3, the in vitro engineered immune cell according to any one of claims 36 to 38, or as The composition of any one of the 39th to 47th patent applications; The measured patient is administered an effective amount of an activating ligand; (d) measuring the amount or activity of IFN-γ in the second biological sample or both. The second biological sample is administered to the vector, immunized a cell or a TSC population, an in vitro engineered immune cell, and an activating ligand are obtained from the patient to be measured, thereby generating a detection amount; and (e) comparing the control amount and the detection amount of IFN-γ, wherein IFN The increase in the amount of gamma, the amount of activity, or the amount of both, compared to the amount of control, indicates that the therapeutic formulation is effective for the patient in need of treatment. -10- 201207108 69.  - a kit comprising: (a) a carrier according to any one of claims 1 to 30, or an immune cell or TSC group according to any one of claims 33 to 35, as claimed in the patent application The in vitro engineered immune cells of any one of the items 36 to 38, or the composition of any one of claims 3-9 to 47, and (b) the activation gene switch body. 70.  For example, the kit of claim 69, wherein the system is RG-115819, RG-115830 or RG-115932. f | 71 - a method for increasing the expression of TNF-α mRNA or the expression of TNF-α protein, the method comprising: (1) producing a vector according to any one of claims 1 to 30, wherein the immunomodulation The agent is TNF-a and wherein the polynucleotide encoding one or more proteins having the function of an immunomodulatory agent further comprises one or more regulatory sequences, and (2) an activating ligand is added, wherein the one or more regulatory sequences Improve the performance of the TNF-a. 72. a use of (a) a carrier for conditionally expressing a protein and (b) a therapeutically effective amount of one or more activating ligands for the manufacture of a medicament for treating a disease or disorder in a mammal in need thereof, wherein the vector comprises a coding gene switch a polynucleic acid, wherein the polynucleotide encoding the gene switch comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a polynucleotide encoding one or more proteins operably linked to a promoter, the promoter being activated via the ligand-dependent transcription factor, wherein the vector is not contained in the cell; and -11 - 201207108 When the ligand is administered, the ligand induces expression of one or more proteins in the mammal to treat the disease. 7 3. The use of the scope of claim 72, wherein the disease is selected from the group consisting of chronic kidney disease, osteoarthritis, tumor, viral upper respiratory tract infection, cat plasma cell stomatitis, cat eosinophilic granuloma, feline leukemia infection, dog Hot infection, systemic fungal infection, cardiomyopathy, mucopolysaccharidosis type VII or infectious disease. 7 4. The use of the scope of claim 73, wherein the infectious disease is selected from the group consisting of bovine respiratory disease, swine respiratory disease, avian influenza, avian infectious bronchitis, bovine spongiform encephalopathy, canine leishmaniasis, chronic wasting disease Classical swine fever, echinococcosis, endemic pneumonia, FIP, foot and mouth disease, lung adenoma, Maedi-Visna disease, mastitis of animals, microsporosis of dogs, contagious sheep Aphthous (animal disease), small ruminant animal disease, pox disease, parrot feather disease, rabies, Mediterranean fever ( brucellosis) or Bang's disease or wave heat, maltese fever 'infectious abortion, epidemic Sexual abortion, Salmonella food poisoning, intestinal paratyphoid fever, bacillary dysentery, pseudotuberculosis, plague, plague fever, tuberculosis, vibrosis, circle disease, Weil's disease (Leptospirosis) or canine hook Spirulina fever, hemorrhagic jaundice (hemorrhagic scutellaria), dairy worker fever (Hart coke), regenerative heat, regenerative heat, spiral heat, vagrant fever, famine fever, Lyme arthritis, Banwo Bannworth's syndrome (Lyme disease), sputum meningococcal polyneuritis, chronic migratory erythema, vibrosis, Escherichia coli, Escherichia coli toxemia, ferrets, swine intestinal edema, intestinal paratyphoid, Staphylococcal gastrointestinal poisoning, staphylococcal gastroenteritis, canine coronavirus-12- 201207108 (CCV) or canine prion enteritis, feline infectious peritonitis virus, infectious gastroenteritis (TGE) virus, Hagerman's red Oral disease (ERMD), infectious hematopoietic necrosis (IHN), Actinobacillus hominis (H. haemophilus) pleuropneumonia, Hansen's disease, streptomycet, sheep fungal dermatitis, false snoring, Hui Whitmore's disease, Francis' disease, deer fever, rabbit fever, O'Hara's disease, Streptomyces fever, Haverhill fever, epidemic joint Erythema, rat bite heat, shipping or transport heat, hemorrhagic sepsis, parrot disease, parrot fever, chlamydia, North American bud disease, Chicago disease, Gilchrist's disease, cat catching heat, Benign lymphoid reticuloendothelial Increased, benign non-bacterial lymphadenitis, bacillary angiomatosis 'bacterial purpura hepatitis, Q fever, Balkan influenza, Balkan grippe, slaughterhouse fever, sputum fever, lung Gram-negative disease, American typhus typhus, typhus typhus fever, blister rickettsial disease, spotted fever in Royal Botanic Gardens, typhus typhus fever, endemic typhus fever, urban typhus , Q money, skin sputum, sputum, sputum, microsporosis, femur, Hong Kong foot, S. skrjab, dimorphic fungus, cryptococcosis and histoplasmosis, benign epidermis Monkeypox, BEMP, simian herpes virus, simian B virus disease, type C lethargy encephalitis, yellow fever, black vomiting disease, Hanta virus pulmonary syndrome, Korean hemorrhagic fever, epidemic nephropathy, epidemic hemorrhagic fever, hemorrhagic Nephritis nephritis, lymphocytic choriomeningitis, Venezuelan equine encephalitis, California encephalitis/La Crosse encephalitis, African hemorrhagic fever, green monkey disease, waterphobia, rabies, infectious hepatitis, epidemic hepatitis, epidemic jaundice , rash, measles, swine flu, equine flu, chicken mites, Newcastle disease, pear larvae, toxoplasma-13- 201207108 disease, African sleep disease, gingival trypanosomiasis, Rhodesia trypanosomiasis, Chaga Sjogren's disease, Chagas-Massa's disease, Chagas disease, E. histolytica, pouch dysentery, cryptosporidiosis, pear-shaped flagellate, cutaneous leishmaniasis: ulceration of gum workers, Nasopharyngeal mucosal leishmaniasis, pianbols, uta and bub a (America), Oriental sore, Oriental sputum (Old World), Baghdad, New Delhi 疖, Bauru ulcer, organ leishmaniasis: kala-azar, microsporidia Disease, stomach nematode, trichinosis, angiostrongosis, eosinophilic meningitis or meningoencephalitis (Vibrio cholerae), abdominal vascular nematode (Costa sinensis), hookworm, board Oral nematode disease, hookworm disease, capillary nematode, malay filariasis, ascariasis, nodular nematode, roundworm, round worm, chicken mites, schizophrenia, schizophrenia, package Insect cyst, echinococcosis, granular echinococcosis (Echinococcus Granulosis), cystic echinococcosis, mites infection, Schistosoma, Burkitt's lymphoma caused by EBV, Lloyd's sarcoma caused by Rous retrovirus, type 8 Kaposi sarcoma caused by herpes virus, adult T cell leukemia caused by HTLV-I retrovirus or hair cell leukemia caused by HTLV-II. 75 . The use of claim 73, wherein the infectious disease is selected from the group consisting of bovine respiratory disease, swine respiratory disease or avian influenza. 76.  For use in claim 73, wherein the tumor is selected from osteosarcoma, leukemia or lymphoma. 77.  The use of any one of the items 72 to 76, wherein the one or more proteins are selected from the group consisting of erythropoietin, ghrelin, osteoproteger, RANKL, RANKL decoy, TNF-α antagonist, IL- 1 antagonist-14-201207108 agent, G-CSF, GM-CSF, IFN-tx, IFN-γ, angiostatin, endostatin, TNF-ct, PP1DCY-LSRLOC, β-uridine or IL-12 . 78.  The use of any one of the items 72 to 77, wherein the one or more proteins are selected from the group consisting of 1 [-1, 1 [-2, 1 [-12, 1 [-3, 11^ 4, IL] -5, IL-7, IL-8, IL-9, IL10R DN or subunits, IL-15, IL-18, IL-21, IL-23, IL-24, IL-27, GM-CSF , IFN-α, IFN-γ, IFN-α 1 , IFN α 2 , IL -1 5 - R-α , CCL3 (MIP-la), CCL 5 (R AN TE S), CCL7 (MCP3), XCL1 ( Lymphocyte Chemokines), CXCL1 (MGSA-a), CCR7, CCL19 (MIP-3b), CXCL9 (MIG), CXCL 1 0 (IP-1 0), CXCL 1 2 (SDF-1), CCL21 (6Ckine &gt; OX40L, 4-1BBL, CD40, CD70, GITRL, LIGHT, b-Prostatin, HMGB1, Flt3L, IFN-β, TNF-a, dnFADD, BCG, TGF-a, PD-L1 RNAi, PD- L 1 antisense oligonucleotide, TGFbRII DN, ICOS-L, S100, CD40L, p53, survivin, p53-survivin fusion, MAGE3, PSA or PSMA. 79.  - (a) a carrier for conditionally expressing a protein and (b) a therapeutically effective amount of one or more activating ligands for use in the manufacture of a medicament for treating a lysosomal storage disease in a mammal in need thereof, wherein the vector comprises a coding a polynucleotide of a gene switch, wherein the polynucleotide encoding the gene switch comprises (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a -15-201207108 nucleotide encoding one or more proteins operably linked to a promoter, the promoter being activated via the ligand-dependent transcription factor, wherein the vector is not contained in the cell; Wherein when the ligand is administered, the ligand induces expression of one or more proteins in the mammal to treat the lysosomal storage disease. 80. For example, the use of the lysosomal storage disease is selected from the group consisting of Pompe's disease/hepatic glucose storage type II, Gaucher's disease (type I, type II). , type III), Fabry's disease, mucopolysaccharidosis type II (Hunter's syndrome), mucopolysaccharidosis type VI (Maroteaux-Lamy's syndrome), mucopolysaccharide Type I, metachromatic white matter dystrophy, neuronal waxy lipofuscinosis or CLN6 disease (atypical late infant, late-onset variant, early juvenile, Finnish variant late infant CLN5, Zhan - (Jansky-Bielschowsky) second disease/late infant type CLN2/TPP1 disease, Kufs/adult NCL/CLN4 disease, northern epilepsy/variant late infant type CLN8, Santavuori-Haltia Ershi/Sp. St. Philip's Syndrome, Type C St. Philip's Syndrome, Type D St. Philip's Syndrome, MPSI Hurler's Syndrome, Niemann-Peel (Niemann-Pick) disease (type A, type B, type C, type D), lack of activator / GM2 ganglioside, alpha-mannose, aspartame, glucosamine, cholesteryl Ester storage disease, chronic hexosaminidase deficiency, cysteosis, Danon's disease, Farber's disease, fucosidase deficiency, galactose sialidase deficiency (Goldberg syndrome), GM1 gangliosides (infant-16-201207108, late infant/juvenile, adult/chronic), I-cell disease/viscosity type II, infancy free saliva Acid storage disease / is SD, juvenile hexosaminidase A deficiency, Krabbe's disease (infancy, late onset), mucopolysaccharidosis (Hurler's multiple malnutrition) Symptoms / viscidosis type III A, Scheie's disease, MPS type I, Heller-Say syndrome, Morquio's disease type A / MPS IVA type, Morki's disease B Type / MPS type IVB, MPS type IX hyaluronidase deficiency, Sly (Sly) disease (MPS type VII), viscid type I / sialic acid deposition, viscidity IIIC , visceral type IV), multiple sulphate lipase deficiency, dense osteogenesis imperfecta, Sandhoff's disease / adulthood / GM2 ganglioside, Sandhoff's disease / GM2 Gangliosides - infant type, Sandhoff's disease / GM2 gangliosides - juvenile type, Schindler's disease, Salla's disease, infantile sialic acid storage disease , Tay-Sachs disease / GM2 ganglioside, Wolman's disease, aspartame glucosamine or saposin. Q 81. The use of the lysosomal storage disease system is selected from the group consisting of Pompe's disease/hepatic glucose storage type, Gaucher's disease (type I, Type II, Type III), Fabry's disease, mucopolysaccharidosis type II (Hunter's syndrome), mucopolysaccharidosis type VI (Maroteaux-Lamy's syndrome), Mucopolysaccharidosis type I or heterochromatic white matter dystrophy. 82. The use according to any one of claims 79 to 81, wherein the one or more proteins are selected from the group consisting of a-galactosidase A, arylsulfatase A, a-glucosidase, b-glucosidase Glucocerebrosidase, ClN6 -17- 201207108 Protein, juvenile-related CLN 3, N-thioglucosamine hydrolase (SGSH), aN-acetylglucosamine, acetaminophen A-glucosamine Base transferase, N-acetylglucosamine-6-sulfate lipase, a_L-iduronidase, arylsulfatase B, acid sphingomyelinase or iduronic acid lipase. 83. - (a) a carrier for conditionally expressing a protein and (b) a therapeutically effective amount of one or more activating ligands for use in the manufacture of a medicament for treating a liver disease in a mammal in need thereof, wherein the vector comprises a multinuclear encoding a gene switch And a polynucleotide comprising the gene coding switch comprising: (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a promoter operably linked to a polynucleotide encoding one or more proteins activated by the ligand-dependent transcription factor, wherein the vector is not contained within a cell; and wherein when the ligand is administered, The ligand induces expression of one or more proteins in the mammal to treat the liver disease. 84. The method of claim 83, wherein the liver disease is hepatitis B. 8 5 The method of claim 83, wherein the liver disease is hepatitis C. 86. The method of any one of claims 83 to 85, wherein the protein is IFN-α. -18- 201207108 87. The method of any one of claims 83 to 85, wherein the protein is a copper blue plasma protein. 8 8. a use of (a) a carrier for conditionally expressing a protein and (b) a therapeutically effective amount of one or more activating ligands for the manufacture of a medicament for treating an ocular disorder in a mammal in need thereof, wherein the vector comprises a multinuclear encoding a gene switch And a polynucleotide comprising the gene coding switch comprising: (1) at least one transcription factor sequence operably linked to a promoter, wherein the at least one transcription factor sequence encodes a ligand-dependent transcription factor, and (2) a promoter operably linked to a polynucleotide encoding one or more proteins activated by the ligand-dependent transcription factor, wherein the vector is not contained within a cell; and wherein when the ligand is administered, The ligand induces expression of one or more proteins in the mammal to treat the ocular condition. 8 9 · If the application of patent application No. 8 8 is used, the eye disease is selected from the group consisting of glaucoma, open angle glaucoma, angle closure glaucoma, and iris-free glaucoma. Congenital glaucoma, juvenile glaucoma, glaucoma caused by crystals, neovascular glaucoma, post-traumatic glaucoma, steroid-induced glaucoma, Sturge-Weber syndrome glaucoma, uveitis-induced glaucoma, diabetic retina Disease, macular degeneration, macular degeneration, choroidal angiogenesis, vascular leakage and/or retinal edema, bacterial conjunctivitis, fungal conjunctivitis, viral conjunctivitis, uveitis, post-corneal sedimentation, macular edema, intraocular lens implantation Post-inflammatory response, grape-19- 201207108 Membrane inflammatory syndrome, retinal vasculitis, sarcoma-like disease, Eales' disease, acute retinal necrosis, Vogt Koyanaki Harada disease, ocular toxoplasmosis, radiation retina Disease, proliferative vitreoretinopathy, endophthalmitis, glaucoma, ischemic optic neuropathy, thyroid associated eyelid disease, orbital pseudotumor, pigment dispersal syndrome (pigmentation glaucoma), scleritis, superficial scleritis choroidal disease, retina Disease (eg cystoid macular edema, centrality) Serous choroidal disease, suspected ocular plasmin syndrome, retinal vascular disease, retinal artery occlusion, retinal vein occlusion, retinopathy of prematurity, retinitis pigmentosa, familial exudative vitreoretinopathy (FEVR), spontaneous polypoid choroidal vasculature Disease, preretinal macula or cataract. The use of claim 88 or 89, wherein the carrier is administered intravitreally. 9 1. The use of any one of claims 72 to 90, wherein the mammal is a human or a non-human animal. 92. The use of any one of the items 72 to 91, wherein the vector is selected from the group consisting of a plastid, an adenovirus, a retrovirus, an adeno-associated virus, a poxvirus, a baculovirus, a vaccinia virus, and herpes simplex. Virus, EB (Epstein-Barr) virus, adenovirus, gemini virus 'cauliflower mosaic virus, liposome, charged lipid (cell transfectin), DNA_protein complex or biopolymer. 9 3 . The use of any one of claims 7 to 92, wherein the vector is an adenoviral vector. 94. For example, in the application of any one of the claims 72 to 93, the gene is related to the EeR-based gene switch in -20-201207108. 9 5. The use of any one of the claims of the invention, wherein the polynucleotide encoding the gene switch comprises a first transcription factor sequence under the control of the first promoter and a second under the control of the second promoter A transcription factor sequence in which proteins encoded by the first transcription factor sequence and the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. 9 6. The use of any one of the claims 72 to 95, wherein the polynucleotide encoding the gene switch comprises a first transcription factor sequence and a second transcription factor sequence under the control of a promoter, wherein the first The transcription factor sequence and the protein encoded by the second transcription factor sequence interact to form a protein complex that functions as a ligand-dependent transcription factor. 97. The use of claim 96, wherein the first transcription factor sequence and the second transcription factor sequence are linked by an EMCV internal ribosome entry site (IRES). 9 8 . The use of any one of claims 7 to 9 of the patent application, wherein the system is based on a ruthenium. 99.  The use of any one of claims 72 to 97, wherein the system is selected from the group consisting of RG-115819, RG-115932 or RG-115830. 100.  The use of any one of claims 72 to 97, wherein the system is amidoxime or oxadiazepine. •twenty one -