CN1865440A - Novel pathogenetic gene mutation of hypertrophic cardiomyopathy and use thereof - Google Patents

Abstract

The present invention relates to some new mutations of pathogenic genes of hypertrophic cardiomyopathy and their application in the treatment and diagnosis of hypertrophic cardiomyopathy.

Description

Novel mutations and uses of genes causing hypertrophic cardiomyopathy

technical field

The present invention relates to some new mutations of pathogenic genes of hypertrophic cardiomyopathy and their application in the treatment and diagnosis of hypertrophic cardiomyopathy.

Background technique

As early as the mid-19th century, the Frenchman A.Vulpian observed and described hypertrophic cardiomyopathy under the microscope, but it was not until the late 1950s that the unique clinical features of hypertrophic cardiomyopathy were systematically discussed. It is usually characterized by asymmetric left and/or right ventricular hypertrophy, can involve different parts of the ventricle, especially the ventricular septum, and the left ventricular volume can be normal or reduced. The typical morphological change is disordered arrangement of muscle fibers. Clinical manifestations often include dyspnea, angina pectoris, syncope, arrhythmia, and sudden death. Hypertrophic cardiomyopathy was considered to be a rare heart disease in the past, but the epidemiological survey conducted by Maron et al. showed that the incidence rate of young people was as high as 0.2% (Maron BJ, Gardin JM, Flack JM, et al. Prevalence of hypertrophic cardiomyopathy in a general population of young adults: echocardiographic analysis of 4111 subjects in the CARDIA study. Circulation. 1995; 92: 785-789). Our echocardiographic Doppler examination of 8080 normal people in nine provinces and cities in China showed that hypertrophic cardiomyopathy is also very common in Chinese, and the number of patients can be as high as 0.08% (Zou Y. et al., Am.J.Med.2004 , 116:14-18). At least 60%-70% of hypertrophic cardiomyopathy is autosomal dominant, which is called familial hypertrophic cardiomyopathy (FHC). Molecular genetic studies have found that mutations in genes encoding sarcomere structural proteins are the fundamental cause of familial hypertrophic cardiomyopathy [Seidman JG, Seidman C. The genetic basis for cardiomyopathy: from mutation identification to mechanistic paradigms. Cell. 2001; ):557-67; Bonne, Carrier L, Richard P. Familial hypertrophic cardiomyopathy: from mutations to functional defects. Cir Res. 1998;83:580-593].

The sarcomere is the basic unit of myocardial contraction and is mainly composed of thick and thin myofilaments. The thick filaments are mainly myosin and myosin binding protein C (MyBPC), the former includes two myosin heavy chains (MHC) and two light chains - essential/basic light chains (MELC, MLC-1) and regulatory/phosphorylated light chains (MRLC, MLC-2). Thin filaments include actin, α-tropomyosin (α-Tm) and troponin complexes (TnT, TnC, TnI). In addition, sarcomere components include titin and so on.

1. Genes causing familial hypertrophic cardiomyopathy

1. At present, it has been confirmed that mutations of more than 160 sites in 13 genes are related to the pathogenesis of FHC, 10 of which are genes encoding sarcomere structural proteins, and most of the mutations are located in these genes (Marian AJ, Roberts R.The molecular menetic basis for hypertrophic cardiomyopathy. J Mol Cell Cardio. 2001; 33: 655670) (Table 1).

Table 1. Occurrence frequency and mutation types of genes causing familial hypertrophic cardiomyopathy protein Gene chromosome location frequency mutation β-MHCcMyBPCcTnTα-TmcTnIMLC-1MLC-2Titinα-Actinα-MHCPKA potassium channel mitochondria MYH7MYBPC3TNNT2TPM1TNNI3MYL3MYL2TTNACTCMYH6PRKAG2KCNQ4MTTI 14q11.2-q1211p11.21q3215q2219p13.2-q13.23p21.2-p21.312q23-q24.32q24.111q14q7q22-q31.11p34 mitochondria ~50% ~20% ~15% ~5% ~5% <5% <5% <5% <5% rare? rare rare 74 missenses, 3 deletions, 1 nonsense mutation 13 missenses, 12 splice sites, 8 insertion/deletion mutations 12 missenses, 1 splice site, 1 deletion mutation 6 missense mutations 7 missense mutations, 1 deletion mutation, 2 missense mutations, 7 missense mutations, 1 nonsense mutation, 1 missense mutation, 5 missense mutations, 1 missense mutation, 1 frameshift mutation, 1 missense mutation (merged WPW) 1 deletion mutation (combined deafness) isoleucine, glycine tRNA mutation

However, these mutations do not include all genetic mutations causing cardiac hypertrophy.

The purpose of the present invention is to discover new mutations to facilitate early diagnosis and prevention of hypertrophic cardiomyopathy.

Contents of the invention

The present invention is based on the discovery that some previously unreported mutations exist in the causative gene of hypertrophic cardiomyopathy. The invention provides isolated nucleic acids containing such mutations. The present invention also provides oligonucleotides useful for detecting these mutations, and methods of detecting mutations using such oligonucleotides. The present invention provides methods for the diagnosis of hypertrophic cardiomyopathy or drug screening or evaluation using such mutations. The invention additionally relates to microarrays.

Detailed ways

In the present invention, the term "sample" refers to any biological sample derived from a human individual, including but not limited to cells, tissue samples such as body soft tissues such as bone tissue, hair, etc., biological fluids such as lymph, blood, saliva , urine and other body excretions. The term "isolated" means not occurring in nature. An "isolated nucleic acid" refers to a nucleic acid that is not contiguous to adjacent 5' and/or 3' flanking sequences in the genome of the individual from which it is derived. Thus, an "isolated nucleic acid" also includes a nucleic acid inserted into a vector, incorporated into the genome of a heterologous cell, or present on its own. The term also includes recombinant nucleic acids linked to other nucleic acid sequences. Such recombinant nucleic acids may be used to produce fusion proteins.

In the present invention, the wild-type gene refers to the normal gene corresponding to the pathogenic gene of hypertrophic cardiomyopathy. Their sequences are known in the prior art and listed eg in the Genbank database. For example, the sequence of the wild-type β-MHC gene is disclosed with Genbank accession number gi: M57965.1, and represented in the present invention as SEQ ID NO: 1; the sequence of the wild-type cMYBPC gene is disclosed with Genbank accession number gi: Y10629, and is disclosed in this invention. It is represented by SEQ ID NO: 2 in the invention; the sequence of the wild-type cTnT gene is disclosed by Genbank accession number gi: X90780, and represented by SEQ ID NO: 3 in the present invention. However, those of ordinary skill in the art are well aware that due to some errors that may exist in the gene sequencing process, the sequence of the wild-type gene will be different from the sequence indicated by the accession numbers listed herein, or the sequence of nucleotide residues will be different. Numbering has changed. At this point, the final sequence will prevail.

The expression method of gene mutation among the present invention is as follows:

"x nt1>nt2" indicates that a nucleotide substitution has occurred in the reference sequence. Where "x" refers to the site where the mutation occurs, which follows the nucleotide residue numbering of the wild-type gene given by the corresponding accession number in Genbank. ">" indicates that a substitution has occurred. "nt1" refers to the nucleotide before the mutation, that is, the nucleotide residue at the corresponding position in the normal gene (wild type gene). "nt2" refers to the mutated nucleotide residue. For example, "4693C>T" in the cTnT gene indicates that a substitution mutation has occurred in the cTnT gene, resulting in a nucleotide residue at position 4693 in the sequence corresponding to Genbank accession number gi:X90780 (SEQ ID NO:3) listed The "C" at the base site is substituted with a "T".

"x1-x2 del nt1...nt2" indicates a deletion at the nucleotide position "x1" to "x2" of the reference sequence, thereby deleting the core of "nt1" to "nt2" nucleotide residues. For example, "14262-14264delAAG" indicates that nucleotides "AAG" from position 14262 to position 14264 are deleted.

"x1^x2 ins nt1...nt2" indicates that "nt1...nt2" nucleotide residues are inserted between the nucleotide positions at "x1" and "x2" of the reference sequence. For example, "18115^18116insGCAAG" indicates that "GCAAG" is inserted between the 18115th and 18116th nucleotide positions.

In one embodiment, the present invention provides a mutated β-MHC gene or fragment thereof, which corresponds to the following position in the sequence (SEQ ID NO: 1) listed in Genbank accession number gi: M57965.1 , containing one or more mutations selected from the group consisting of: 10353G>A, 10429A>C, 10895A>T, 11811A>C, 13653A>C, 14449T>C, 14452G>A, 14557G>A.

In one embodiment, the present invention provides a mutated cMYBPC gene or a fragment thereof, which contains a gene selected from the group consisting of One or more mutations in the following group: 4826G>A, 8697G>A, 8887G>A, 12101G>A, 13261G>A, 14262-14264delAAG, 14364delG, 18115^18116insGCAAG.

In one embodiment, the present invention provides a mutated cTnT gene or a fragment thereof, which contains selected One or more mutations from the following group: 4693C>T, 4769G>A.

In a further embodiment, the mutated gene or fragment of the present invention also contains additional mutations, including any known or future known mutations or nucleotide polymorphisms.

The present invention also relates to mRNA or cDNA or encoded protein derived from the above mutant gene or fragment.

The present invention also relates to nucleic acids, such as oligonucleotides, which can specifically hybridize to any of the above-mentioned mutant genes or fragments or mRNA or cDNA derived therefrom or their complementary sequences, but cannot hybridize to the corresponding wild-type gene sequence or its complementary sequences. Nucleotides, which can be used as probes or primers to specifically detect the above mutations in human individuals. It is clear to those skilled in the art that the probe generally has a sequence that is complementary to the sequence of the target nucleic acid and specifically hybridizes to the nucleic acid containing the mutation to be detected but not to the nucleic acid having the wild-type gene sequence. The oligonucleotide is generally located around the mutation site, and specifically detects the presence or absence of the above mutation in the sample. Preferably, said nucleic acid or its complement contains any one or more mutations as defined above. As an example, the oligonucleotide primer or probe can be designed such that the mutation site to be detected is located at its 5' or 3' end or in the middle, or the mutation site to be detected is located at 5' to the complementary sequence of the primer or probe. ' or 3' end or in the middle. In the context of the present invention, the term "specifically detects" means that the oligonucleotide hybridizes only to nucleic acids comprising the mutations defined above or their complements, but not to nucleic acids having wild-type gene sequences or their complements, respectively. nucleic acids hybridize.

When multiple nucleotides are involved in the mutation site, the oligonucleotide probe or primer can be designed so that the whole or part of the nucleotide sequence of the mutation site or its complementary sequence is included in the nucleoside of the probe or primer. acid sequence. Thus, it is possible to use the oligonucleotides thus prepared as probes to determine the presence or absence of mutations by hybridization or the presence or absence of differential hybridization, or as primers by amplification followed by detection (by e.g. sequencing, restriction enzyme digestion). etc.) the presence of mutations.

Oligonucleotide probes or primers are generally greater than about 10 bp in length, such as 10-100 nucleotides, preferably, 15-75 nucleotides in length, more preferably 18-50 nucleotides in length, especially preferably 20-35 nucleotides, most preferably 23-30 nucleotides. Those skilled in the art can easily design suitable oligonucleotide probes or primers for the detection of mutation sites. As for its preparation, it can be carried out by methods well known to those skilled in the art, including but not limited to, for example, chemical synthesis. Preferably, the probe/primer is detectably labeled to facilitate detection. Labels can be, for example, biotin, fluorescent dyes, radioactive isotopes, etc., but are not limited thereto. They are known in the art and can be attached to probes/primers by conventional methods. The nucleic acid may be single-stranded or double-stranded.

Primers can be used to extend nucleic acid sequences by adding free nucleotides, as is well known in the art. Hybridization refers to selective binding to a nucleic acid sequence under stringent conditions. Herein, stringent conditions are conditions that allow primers to hybridize only to the mutation-containing nucleic acid sequence or its complement, but not to the wild-type nucleic acid sequence or its complement. According to the primers used, those of ordinary skill can determine the corresponding stringent hybridization conditions through routine experiments. Numerous prior art references are available in this regard, including, for example, Sambrook et al., Molecular Cloning Laboratory Handbook, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

The present invention also relates to mutation detection using primers designed to, upon amplification, result in a fragment comprising the mutation to be detected. The primers are generally located around the mutation site or within the range of 1000bp, preferably 500bp, more preferably 250bp, especially preferably 200bp, most preferably 100bp in the 5' or 3' direction of the mutation site, and its length is generally greater than about 10bp, preferably 15-100bp , more preferably 18-75bp, especially preferably 20-50bp, most preferably 25-40bp.

It is also possible to link some additional nucleotide sequences at the 5' and/or 3' ends of the oligonucleotide sequences of the invention. Such oligonucleotides are also within the scope of the invention.

Using such oligonucleotides, the presence or absence of such mutations in a human individual can be detected by amplification and subsequent analysis of genetic material from said human individual. For example, the amplification product can be subjected to agarose gel electrophoresis, followed by staining with ethidium bromide, SYBR green solution, or the like, thereby detecting the amplification product as a band (DNA fragment). Thus, a gene fragment including a mutation site can be detected as a DNA fragment. In addition to agarose gel electrophoresis, polyacrylamide gel electrophoresis or capillary electrophoresis can also be performed. Further, the amplified products can be isolated and then analyzed for the presence or absence of mutations by sequencing or restriction enzyme digestion. It is also possible to perform PCR using primers previously labeled with a substance such as a fluorescent dye, and to detect the amplified product. A detection method that does not require electrophoresis can also be used; in this method, the amplification product is bound to a solid support such as a microplate, and the DNA fragment of interest is detected by means of fluorescence, enzyme reaction, and the like.

The present invention provides a method for detecting the above-mentioned mutations, comprising the detection of biological samples using the oligonucleotide probes or primers described above, or using a primer pair designed to obtain a fragment containing the mutation after amplification. Steps for biological samples and testing. In the detection, biological samples of the individual to be tested are usually used, such as blood thereof, but not limited thereto. In one embodiment, the biological sample is used directly for detection. For example, using the above-mentioned nucleic acid as a primer, a fragment containing the mutation site to be detected is amplified by PCR. In another embodiment, but not limited to, genetic material, such as genomic DNA, is first isolated from the biological sample prior to detection. Detection can be routinely performed by methods such as nucleic acid hybridization, PCR method, sequencing method, restriction fragment length polymorphism assay, etc., but is not limited to these methods. The conditions are known to those of ordinary skill in the art, or can be determined quite easily. Preferably, a DNA fragment encoding a mutation site to be detected is amplified with a DNA polymerase using the oligonucleotides prepared as described above as primers. Alternatively, probes prepared as described above are hybridized to template DNA, thereby detecting those DNAs with mutations of interest. Template DNA can be prepared according to conventional methods, such as cesium chloride gradient centrifugation, SDS cleavage, or phenol/chloroform extraction.

The present invention also relates to a microarray containing the above-mentioned oligonucleotide probes or primers. The oligonucleotide can be immobilized on a solid support. In one embodiment, the microarray is in the form of a DNA chip, gene chip, microchip, bead array, or the like. The DNA chip arranges and immobilizes a variety of DNA probes on a substrate such as a glass slide, and then performs hybridization of labeled DNA on it, and distinguishably detects complete matches and nucleotide mismatches by detecting the label signals on the probes, Nucleic acid is thereby detected. Conventional methods for the preparation of chips are known to those skilled in the art.

The present invention also relates to a kit that can be used to detect the above-mentioned mutations, including one or more relevant diagnostic primers or probes and/or oligonucleotide primers specific to the mutant gene, such as the oligonucleotides described above acid probe or primer. Also included in the kit are containers, instructions for use, buffers, enzymes, and/or other reaction components. Preferably, a positive control and a negative control are also included in order to further confirm the detection results. In one embodiment, the kits of the invention comprise any and all of the component enzymes or components for a predetermined assay. Examples of these components include, but are not limited to, oligonucleotides, polymerases (such as Taq polymerase), buffers (such as Tris buffer), dNTPs, control reagents (such as tissue samples, target oligonucleotides for positive and negative controls) nucleotides, etc.), labeling and/or detection reagents (fluorescent dyes such as VIC, FAM), solid supports, instructions, illustrative charts and/or product information, inhibitors, etc. The kit of the present invention may be a partial kit comprising only some of the necessary components. In this case, the user can provide the remaining ingredients. Kits of the invention may comprise two or more separate containers, each containing a portion of the components to be used. For example, in a kit, the first container contains the enzyme and the second container contains the oligonucleotide. Specific examples of enzymes include structure-specific cleavage enzymes in appropriate storage buffers or containers.

The present invention also relates to a method for diagnosing hypertrophic cardiomyopathy or determining the prognostic risk of hypertrophic cardiomyopathy, comprising the step of detecting one or more of the following mutations in an in vitro sample of an individual: the following mutations in the β-MHC gene: 10353G>A, 10429A>C, 10895A>T, 11811A>C, 13653A>C, 14449T>C, 14452G>A, 14557G>A; the following mutations in the cMYBPC gene: 4826G>A, 8697G>A, 8887G> A, 12101G>A, 13261G>A, 14262-14264delAAG, 14364delG, 18115-18116insGCAAG; or the following mutations in the cTnT gene: 4693C>T, 4769G>A. Preferably, the detection step is performed according to the mutation detection method described above. For example, the nucleic acid fragment containing the mutation site to be detected is first subjected to PCR amplification using oligonucleotides located upstream and downstream of the mutation site. Detection can be performed by, for example, PCR method, sequencing method, restriction fragment length polymorphism and the like.

The present invention also relates to a method for screening or evaluating drugs using the above-mentioned mutant gene as a target, including, for example, administering one or more drugs to cells, tissues, organs or animal individuals carrying the mutation, and evaluating one or more indications thereof Indicator of hypertrophic cardiomyopathy status to indicate disease improvement status. Preferably, the cells, tissues, organs or individual animals are obtained by gene knockout (gene knockout) or knockin (gene knockin) or transgenic methods. The methods of gene knockout or knockin or transgene are well known to those skilled in the art, including, for example, designing two flanking fragments homologous to the endogenous gene. In the case of gene knockout, a mutation intended to inactivate an endogenous gene in the body is contained between the two homologous fragments, so that the corresponding endogenous gene can be inactivated after being introduced into the body. In the case of gene knock-in, the nucleic acid intended to be introduced into the endogenous gene in the body is contained between the two homologous fragments, so that after the introduction into the body, the target nucleic acid can be inserted into the predetermined site. The subject may be an individual, such as a mouse. Alternatively, the object of operation may also be cells, such as human cardiomyocytes. In this way, drugs can be screened or evaluated at the individual or cellular level.

The present invention is illustrated below by means of examples. These examples are exemplary and they do not limit the scope of the invention in any way.

Example

The extraction of embodiment 1 genome DNA

1 Selection of research objects

The subjects of the study were 100 patients with hypertrophic cardiomyopathy who were clearly diagnosed in the outpatient department or hospitalized in the Fuwai Cardiovascular Hospital of the Chinese Academy of Medical Sciences. There was no blood relationship through family investigation. Among them, there were 51 probands in a family with hypertrophic cardiomyopathy, 356 related family members, and 49 sporadic cases. 120 normal study controls were healthy people matched in sex and age. The selected candidates fill in the "Molecular Genetics Research Case Investigation Form", which includes: patient contact information, personal general conditions, clinical symptoms, physical examination results, family history investigation, laboratory examination, electrocardiogram (Holter if necessary), chest X-ray report, ultrasound Cardiogram report (cardiac MRI or left ventricle and coronary angiography results if necessary). Build a data database.

Diagnostic criteria for hypertrophic cardiomyopathy (published by WHO/ISFC in 1996)

(1) Basic criteria: unexplained cardiac left ventricular wall thickness greater than or equal to 13mm diagnosed by echocardiography; or apical hypertrophy or right ventricular hypertrophy.

(2) Reference standard: left ventricular high voltage on electrocardiogram, with/without pathological Q wave and ST-T changes in leads II, III, avF, avL and V ₄ V ₅ .

(3) Exclusion criteria: long-term hypertension, ischemic cardiomyopathy and other primary diseases that can cause ventricular hypertrophy.

(4) Determination criteria for familial hypertrophic cardiomyopathy: There is a family genetic tendency of cardiomyopathy, that is, multiple patients in the patient (proband) family have the disease, and it conforms to the characteristics of autosomal dominant inheritance.

2 Isolation of genomic DNA

Take 5ml of blood from the patient's cubital vein, anticoagulate with EDTA (0.5M), and centrifuge at 2500rpm for 10min. Carefully draw the upper layer of plasma, and use 1.5ML Eppendorf tubes. Add 3 times the volume of hemolysate (NH ₄ Cl 8.29g, KHCO ₃ 1.0g, EDTA-Na ₂ 0.03734g, add double distilled water to 1000ml, adjust the pH to 7.4) into the blood cells, shake well, and place in ice bath for 15min. Centrifuge at 2500rpm for 10min, discard the supernatant. Add 10ml of hemolysate to the cell pellet, shake well, and ice-bath for 15min. Centrifuge at 3000rpm for 10min, discard the supernatant to obtain white blood cells, which can be directly used to extract genomic DNA, or frozen at -80°C for later use.

Genomic DNA was extracted as follows: take white blood cells, add 4.5ml SE (NaCl 4.383g, EDTA-Na ₂ 9.306g, add double distilled water to 1000ml, adjust the pH value to 8.0), vigorously blow and mix. Add 0.5 ml of 10% SDS along the tube wall, then add 50 μl of protease E (100 mg/ml, MilliQ water), shake gently from head to tail for 10 min, mix well, and then place in a water bath at 37°C overnight. Take it out the next day, add 3ml of saturated NaCl along the tube wall, shake it quickly by hand for 15s to denature and precipitate the protein, and then centrifuge at 4000rpm for 10min. Take the supernatant, transfer it to another tube, add an equal volume of chloroform, shake for 10 minutes, and mix well. Centrifuge at 2500rpm for 10min. Take the supernatant, transfer it to another tube, add an equal volume of chloroform, shake for 10 minutes, and mix well. Centrifuge at 2500rpm for 10min, take the supernatant, transfer it to a 50ml tube, add 3 times the volume of absolute ethanol, shake gently from head to tail, and see DNA flocculent precipitate. Pick the DNA to the wall of the tube and wash it twice with 75% ethanol. DNA was picked into 0.5ml tubes and lyophilized. Finally, 300 μl of TE (10 mM Tris-HCl, 1 mM EDTA, adjusted to pH 8.0) was added to dissolve the DNA, and the OD value was measured and quantified by an ultraviolet spectrophotometer, and stored at 4°C.

Example 2 PCR method for screening mutations in the β-myosin heavy chain (B-MHC) gene

The primers and amplification conditions used are shown in Table 1 below (genome sequence is from Genebank database, accession number is M57965):

Table 1 Exon PCR fragment length Primer PCR reaction conditions Exon3Exon4Exon5Exon6Exon7Exon8Exon9Exon10Exon11Exon12Exon13Exon14Exon15Exon16Exon17Exon18Exon19Exon20Exon21Exon22Exon23Exon24Exon25Exon26Exon27 245bp248bp200bp146bp244bp240bp223bp226bp229bp398bp267bp213bp232bp438bp180bp474bp271bp298bp285bp300bp400bp282bp221bp151bp420bp F 5'act cca ggc aca gcc atg gg 3'R 5'gtg gac tct cac atc agc ct 3'F 5'gaa agc cca ggc tga cag ga3'R 5'ggg tgg aca tgg atg gag ca 3'F 5' gca act ggc aag tca ctg ct 3'R 5'ctc tca cct gtc agc atg ta 3'F 5'gag aag ccc cac gag agc at 3'R 5'tgg agg ctg ggg atc agg gag 3'F 5'ctg att tga ggc ttg ctg gt 3'R 5'aag aag gag gca ggt gag ag 3'F 5'cgc aga agg gag gga gaa ga 3'R 5'agt ggt gat gag ttg ggg ga 3'F 5'gga ctt gga ctg gtg gag ga 3'R 5'ggg gag aga gag aga gga ca 3'F 5'att aat ttt cct tgt gcc ca 3'R 5'caa gca gag ggg acc agg tt 3'F 5'cct cca tga gag ccg ggg gc 3'R 5'ctg ctt ttg gac ccc tgt tt 3'F 5'gga tct cac tta ccc atc at 3'R 5'ggg agg ctg agg cag gag a 3'F 5'tta cag gca tga acc aca cac c 3'R 5'gtg aac ttg aaa act ctc atc cc 3'F 5'gtc tct cct cca cct tgc ag 3'R 5'cga gtg agt gat tgt tct cc 3'F 5'acc cac ttt ctg act gct cc 3 'R 5'tgg taa ggc caa aga ggc ac 3'F 5'ttg acc ata gag cag aat cc 3'R 5'ctc aga acc ttg gga gaa tc 3'F 5'gct gtg atc tta ctc aca cc 3'R 5'tgg gca gat ggg gag cca ag 3'F 5'tct ctc ctc ttt ccc ttc tgt ctc 3'R 5'tgc ctt aca cat cac cct cat tac 3'F 5'aat cag tga caa agc cag ga 3'R 5'gca gat ccc att ccc atc ag 3'F 5'cct gat ggg aat ggg atc tg 3'R 5'atg att aca aca gga aaa gc 3'F 5'tcc taa ggt aat ccc cac ca 3'R 5' ctt ttt ttc ctg aca ctg cc 3'F 5'gca gag gct cag cac tcc tt 3'R 5'gcc tca ccg cct gca ctt gg 3'F 5'ccc tcc tat ttg agt gat gtg c 3'R 5'ggt cag tat ggt ctg aga gtc c 3'F 5'aca gat ggc acc aag ctg gt 3'R 5'gat aga cat ggc ata tca ag 3'F 5'ggc tcc ttc tct cta cca gc 3'R 5'tgc ttg tac tgt tat ggg ct 3'F 5'acg agt ctc cct tac ctc ac 3'R 5'cgt ggg tgt cca gac ctc ac 3'F 5'gtg tct gac cag gca ctc at 3'R 5'agg ccg agc aga gcc tac ct 3' 95℃5', 94℃30", 61.5℃30", 72℃30"(35cycles);72℃7'95℃5',94℃30",60℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",58℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",60℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",56℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",61℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",59.2℃30",72℃30"(35cycles);72℃7'95℃5',94℃30", 55 ℃30", 72℃30"(35cycles);72℃7'95℃5',94℃30",64℃30",72℃30"(35cycles);72℃7'95℃5', 94℃ 30", 53℃30", 72℃30"(35cycles);72℃7'95℃5',94℃30",61℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",61℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",60℃30",72℃30"(35cycles); 72℃7 '95℃5', 94℃30", 55℃30", 72℃30"(35cycles);72℃7'95℃5',94℃30",63℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",68℃30",72℃50"(45cycles);72℃7'95℃5',94℃30",57℃30", 72℃30 "(35cycles);72℃7'95℃5',94℃30",55℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",58℃30" , 72℃30”(35cycles); 72℃7’95℃5’, 94℃30”, 60℃30”, 72℃30”(35cycles); 72℃7’95℃5’, 94℃1’, 59.5℃40", 70℃1'(35cycles);70℃10'95℃5',94℃30",57.4℃30",72℃30"(35cycles);72℃7'95℃5', 94 ℃30", 60.6℃30", 72℃30"(35cycles);72℃7'95℃5',94℃30",62℃30",72℃30"(35cycles);72℃7'95℃5', 94°C 30", 60°C 30", 72°C 30"(35cycles); 72°C 7'

Prepare the basic PCR reaction system (50 μl) as follows:

Primer (concentration 20pmol/μl) 1μl

10×buffer 5μl

TaqDNA polymerase (2u/μl) 1μl

dNTP(10mM) 1μl

Human Genomic DNA 1μl

ddH ₂ O 41μl

According to the PCR conditions in Table 1, the genomic DNA of each patient or control was amplified by PCR, and the PCR reaction products were purified before sequencing according to the following steps:

(1) Mix 50 μl of the PCR reaction product with 6 times its volume of a mixed solution of absolute ethanol and 5M ammonium acetate (5:1), and place at -80°C for more than 30 minutes.

(2) Centrifuge at 14,000 rpm at 4°C for 15 minutes, and discard the supernatant.

(3) Add 200 μl of 75% ethanol, centrifuge at 14000 rpm for 5 minutes at 4° C., and discard the supernatant.

(4) After room temperature or lyophilization, add 20 μl TE to dissolve.

(5) Aspirate 2 μl for spotting and quantification.

The sequencing reaction was performed as follows:

(1) Reaction system (20μl)

BigDye ^™ 4 μl

BigDye ^™ Buffer 2μl

Primers (same as PCR amplification primers) 10pmol/μl

(2) Reaction conditions (depending on the annealing temperature of the primer):

(3) Purification of sequencing products:

Add 25 μl ammonium acetate and 125 μl ethanol to 20 μl product, place at -70°C for 1 hour, centrifuge at 14,000 rpm for 10 minutes, discard the supernatant, add 100 μl of 75% ethanol, centrifuge at 14,000 rpm for 5 minutes, discard the supernatant, bake at 95°C for 1 minute After drying, add 2μl loading buffer, mix well, denature at 94°C for 5 minutes, and then quickly put it on ice.

(4) The purified sequencing reaction product was sequenced on the ABI PRISM377 automatic sequencer.

(5) Use GeneJockey, AssemblyLIGN ^TM , Sequencing Analysis 3.4 software to read and check the sequencing map, and compare with the published gene sequence. The gene mutations thus screened are listed in Table 5.

Embodiment 3 PCR method screens the mutation in the cardiac troponin T (cTnT) gene

The primers and amplification conditions used are shown in Table 2 below (genome sequence is from Genebank database, accession number is AY044273):

Table 2 Exon PCR fragment length Primer PCR reaction conditions Exon2Exon3, 4Exon5, 6Exon7Exon8, 9Exon10, 11Exon12, 13Exon14Exon15, 16 203bp272bp596bp292bp682bp1357bp743bp315bp667bp F 5'tcg ctg atc tct gta caa cgg ggg-3'R 5'ggt gtc agg gca gcg gcg g-3'F 5'atg tgt gtg cag gcg tgt gtg ttt-3'R 5'cca ggg ctc cca gga ttt cca-3'F 5'tct ctg ctc tgg gtt ctg cct gat a-3'R 5'att ctc ctc caa agc tgc tgt ga-3'F 5'gcc ccc gct gcc ttc att c-3'R 5'cac cat act gca ccc tgt tcc atc a-3F 5'ggg cag tgc tgg aag att ctc tag g-3'R 5'gag gaa ggc tgt ctg gag gag gt-3'F 5'gtc cgt gga gct ggt tga aag tg- 3'R 5'aat ctc ttc ctg gga cct gac cta aa-3'F 5'ctt tgt ctt cct gcc ttc tct ctg tgt-3'R 5'cag aaa gga gtt ggc ccg acc-3'F 5'gga agg agg gct ggg atg ctg-3'R 5'ctg aag ggg gct gtt ggg ga-3'F 5'cac tca gcc ccc ttc tcc cca-3'R 5'cca gct ccc cat ttc caa aca-3' 95℃5', 94℃30", 61℃30", 72℃30"(35cycles);72℃10'95℃5',94℃30",61℃30",72℃30"(35cycles);72℃10'95℃5',94℃30",61℃30",72℃30"(35cycles);72℃10'95℃5',94℃30",63℃30",72℃30"(35cycles);72℃10'95℃5',94℃30",63℃30",72℃1'(35cycles);72℃10'95℃5',94℃30",61℃30",72℃1'(35cycles);72℃10'95℃5',94℃30",61℃30",72℃1'(35cycles);72℃10'95℃5',94℃30", 61 ℃30", 72℃30"(35cycles);72℃10'95℃5',94℃30",63℃30",72℃1'(35cycles);72℃10'

A PCR reaction basic system (50 μl) basically similar to Example 2 was prepared, except that the primers of this example were used instead.

The genomic DNA of each patient or control was amplified by PCR according to the PCR conditions in Table 2, and the amplified products were separated and sequenced by conventional methods. The gene mutations thus screened are listed in Table 5.

Example 4 PCR method for screening mutations in cardiac troponin I gene

The primers and amplification conditions used are shown in Table 3 below (genome sequence is from Genebank database, accession number is X90780):

table 3 Exon PCR fragment length Primer PCR reaction conditions Exon3Exon4Exon5Exon6Exon7Exon8 128bp163bp239bp251bp287bp239bp F 5'ggg tcc cca ctc ctc cta-3'R 5'tcc ggc gcc tgt act ctg-3'F 5'tgc tgg ggg tgt ctt gag gt-3'R 5'cac ctg cct gct ctt tcc ca-3' F 5'gag ggt tta gaa ggg cag agg-3'R 5'ccg gga cta gaa acc tcg ca-3'F 5'ggt ctc cct gtt ttt ggt tcc c-3'R 5'tgg gtc tcc tgg gat gtg ca -3F 5'atg gag gag ttg ggt gtg cg-3'R 5'aca gcc ctt ccc ctc agc at-3'F 5'aga ctg gag agg aag aag aga ccc-3'R 5'tcc tgc cta agc cct ggg taa t-3' 94℃5', 94℃30", 60℃30", 72℃30"(35cycles);72℃7'94℃5',94℃30",63℃30",72℃30"(35cycles);72℃7'94℃5',94℃30",60℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",60℃30",72℃30"(35cycles);72℃10'95℃5',94℃30",64℃30",72℃1'(35cycles);72℃10'95℃5',94℃30",60℃30", 72°C 1'(35cycles); 72°C 10'

A PCR reaction basic system (50 μl) basically similar to Example 2 was prepared, except that the primers of this example were used instead.

The genomic DNA of each patient or control was subjected to PCR amplification according to the PCR conditions in Table 3, and the amplified products were separated and sequenced according to the method in Example 2. The gene mutations thus screened are listed in Table 5.

The PCR amplification of embodiment 5 cardiac myosin binding protein C gene (cMYBPC)

The primers and amplification conditions used are shown in Table 4 below (genome sequence is from Genebank database, accession number is Y10629):

Table 4 Exon PCR fragment length Primer PCR reaction conditions Exon1Exon2Exon3-5Exon6Exon7-9Exon11.12exon13exon14-17exon18exon19.20exon21.22exon23exon24.25exon26exon27exon28.29exon30-.32exon33.34 145bp550bp1016bp344bp674bp594bp315bp915bp537bp537bp623bp525bp619bp567bp275bp770bp996bp605bp F 5'atg gtg agt agc ctg gtg tga c 3'R 5'ccc cct caa gaa ctc ctc ct 3'F 5'tgt cct cct ctg aga aat gaa cag 3'R 5'cag caa agg caa gaa agt gtg a 3 'F 5'gtg gag ggg tct ctg gtt agt 3'R 5'tct gtg tgc ctt gtg cct tct3'F 5'ggg tgc gga gcc ttg tct3'R 5'gcc ttg ggg ttt ctc tca cac3'F 5'cag ggc agg gct tct caa a3'R 5'gac aga caa gga aac caa ctc aga3'F 5'ggg tgg ggt cca ggt ctt t3'R 5'atg ccc tct cct ctc ctg tgt3'F 5'gcg gca cag agg gga ttg3' R 5'ggg cag gag gca agg cta t3'F 5'tct cgg ggc tca ctt cct c3'R 5'gtc acc tca gca tcg tca ttt tag3'F 5'cag cct aaa cct cat cct tct ca3'R 5'tgt ttc ctg tgt ctc tct ctg t3'F 5'cac ttt tac gct gcc tac ctc tg3'R 5'atc ctt gct ttt ccc tct gtg a3'F 5['tgg ctc tcc cgt ttc tct ga3'R 5'agc aac aca ccc gac caa gt3'F 5'agt cgg tgc cca gag atg aat3'R 5'cag cag tgt cgc agg aaa tct3'F 5'ctg tgg cgg tta gtt gga gtg3'R 5'agg atg aaa gga gac tgt3'F 5 'act tga act cgg gag gca gaa3'R 5'ccc tct ctg cac ttt ttc cct a3'F 5'cag tgg gag tgg ggt gtc ag3'R 5'ggt gtc aat ggc ggg tct t3'F 5'agg gag gtt gga gct gtg ga3'R 5'tgg gga gga ctt ggc ttg t3'F 5'gga ccc aac cca cag ctc ac3'R 5'agg ctg ggg aga gga ctg c3'F 5'gga ccc ccc gag tag aaa ca3'R 5 'gcc cct aca gcc tcc cat tt3' 95℃5', 94℃30", 58℃30", 72℃30"(38cycles);72℃7'95℃5',94℃30",62℃30",72℃40"(38cycles);72℃7'95℃5',94℃30",62℃30",72℃40"(35cycles);72℃7'95℃5',94℃30",62℃30",72℃30"(35cycles);72℃7'95℃5',94℃30",63.9℃30",72℃40"(35cycles);72℃7'95℃5',94℃30",62.5℃30",72℃40”(39cycles); 72℃7’95℃5’, 94℃30”, 56℃30”, 72℃30”(38cycles); 72℃7’95℃5’, 94℃30”, 63 ℃30", 72℃90"(35cycles);72℃7'95℃5',94℃30",58℃30",72℃40"(36cycles);72℃7'95℃5', 94℃ 30", 58℃30", 72℃40"(38cycles);72℃7'95℃5',94℃30",56℃30",72℃40"(35cycles);72℃7'95℃5',94℃30",53℃30",72℃50"(36cycles);72℃7'95℃5',94℃30",58℃30",72℃40"(38cycles); 72℃7 '95℃5', 94℃30", 55.5℃30", 72℃30"(35cycles);72℃7'95℃5',94℃30",60℃30",72℃40"(40cycles);72℃7'95℃5',94℃30",62℃30",72℃50"(40cycles);72℃7'95℃5',94℃30",66.5℃30", 72℃120 "(40cycles);72℃7'95℃5',94℃30",56.3℃30",72℃50"(35cycles);72℃7'

A PCR reaction basic system (50 μl) basically similar to Example 2 was prepared, except that the primers of this example were used instead.

The genomic DNA of each patient or control was subjected to PCR amplification according to the PCR conditions in Table 4, and the amplified products were separated and sequenced according to the method in Example 2. The gene mutations thus screened are listed in Table 5.

Example 6 Restriction Fragment Length Polypeptidic (RFLP) Detection

Due to the base change at a certain position of the gene, a specific enzyme cutting site that can be recognized by a restriction endonuclease is destroyed or established. When the sample is treated with a specific restriction endonuclease, because of its different Digestion fragments with different lengths can show different migration speeds during gel electrophoresis. Based on this principle, gene mutation screening of family members of patients with known mutation sites can be carried out. Promega enzyme digestion kit was used in this detection, and it was carried out according to the attached product instructions.

Table 5 Screening results of β-MHC, cMYBPC, cTnT and cTnI gene mutations in 100 patients with hypertrophic cardiomyopathy: disease-causing gene Intragenic position Nucleotide change amino acid changes Transcript changes frequency Patient percentage (%) MYH7 (β-MHC) gi: M57965.1 exon 3exon 5exon 14exon 14exon 15exon 16exon 18exon 18exon 19exon 20exon 20exon 22exon 22exon 22exon 23exon 23 5797 C＞T6739 G＞A10353 G＞A10429 A＞C10895 A＞T11811 A＞C10600 C＞T10601 G＞A13463 G＞A13653 A＞C13673 G＞C14449 T＞C14452 G＞A14557 G＞A15373 G＞A15391 G＞A A26VR143QG425RK450TI511FK615QR663CR663HR719QQ734PG741RM822TG823ER858HE924KE930K missense missense missense missense missense missense missense missense missense missense missense missense missense missense missense 4211111411111121 twenty four MYBPC3 (cMYBPC) gi: Y10629 exon 6exon 7exon 15intron 15exon 21exon 23exon 25exon 25exon 29 4295 G＞A4826 G＞A8697 G＞A8887 G＞A12101 G＞A13261 G＞A14262-14264delAAG14364delG18115^18116insGCAAG E258KG263RG416SR668HG758D missense missense missense splicing missense missense codon deletion frame shift 211111111 13 TNNI 3 (cTnT) gi: X90780 TNNT2 (cTnI) gi: AY044273 exon 7exon 7exon 10 4693 C＞T4769 G＞A14035 T＞C R145WR170QR130C missense missense missense 211 31

Note: Those with heavy colors are all new mutations, a total of 19.

2. MHC: G425R, Q734P, M822T, G823E; cMYBPC: G263R, 8887 G>A (intron15), R668H, G758D, 18115^18116 insGCAAG (exon 29); cTnT: R145W and other mutations were found in family patients, and hypertrophy Cardiomyopathy showed a co-segregation trend.

3. No mutations of the above sites were found in 120 cases of normal controls.

The above-mentioned embodiments are only for illustration, not limitation of the present invention. It will be apparent to those skilled in the relevant art that other suitable modifications and variations can be made to what is described herein and that such modifications and variations can be made within the scope of the invention or any embodiment thereof. Such modifications and changes all fall within the protection scope of the present invention.

sequence listing

<110> Fuwai Cardiovascular Hospital, Chinese Academy of Medical Sciences

<120>New mutations of hypertrophic cardiomyopathy gene and its application

<130>idc040150

<160>4

<170>PatentIn version 3.2

<210>1

<211>28438

<212>DNA

<213>β-MHC gene

<220>

<221>misc_feature

<222>(11380)..(11380)

<223> n is a, c, g, or t

<400>1

gatctatctg gtccagacca tcctttgaac caagtggact tagtggggct gaaggaggtg 60

tggggacaa gaacctatgg cagggcaacc ggccctcctg tctgcagcct ggcttcctca 120

attttgtgtt cctaacctaa gagctgccat tcagtgaggg cccacagtgt gtctgctgtg 180

tgtagactct gcttgtgcaa tctctaatgg cctgagtagc cctggaaaca aaatgtggct 240

atccccatgt tacagatgac aaaactggct caaagcaatt atatgactga cgtagaccac 300

acagctagaa aatggaatca ctgggtttca aagtggattt ttgtgactga ctccaaagct 360

gaccagagtg ctcatcctgt atgcttctga ggcctctggc cctccagacc ggtcactccc 420

cacaaagtct cccttttggt gagttgtgtc cctccaaaca ctccctgaag gcaagatcaa 480

cctgccctaa ctaccccaaa gtgaactagg ccagccccaa gcctgtctcc ggccctccat 540

ctctctcttt gttcacatcc atcagaaagc agagacctag agtagggaat gagagggggt 600

gcacaagtac aaaagatttt gaagcagcag gaagacgaaa gaagtacaag aaggagcagc 660

tgtttgtaaa agggaaaatc tgagacaggc agaaagctca gctccctaat cctaggctcg 720

tgtggagcca gggcagggtc ccccaaggtc agggattctg ggggcaggcg gatgggagag 780

tcagcctcca ctcctgggtg gctcccctct gccctgtgca caaacaacga gtgcctcatt 840

cctcccctgc accaagtgcc ctgcctcttc ctgccccgtt tcttccttcc ctttgggtac 900

cggatctttt ttctgattcc cctctctcca tgctgacatc gcttctaatt cgccctggtg 960

tctccctccc tcacaattac tgggagtaat tttagcaact tttcctctga actaccgctg 1020

gcgtgttcac agcaaatggc ccctgttttg gcttgacact tgatcagggt ggtgacagag 1080

tgcagacaaa gccggcacaa tgcggggagg ggaggaccgg ggcagatcac cttcctttca 1140

cttcagggcc ttatttggtc taccttgccg aagcggccct ccttccagcc tctctttat 1200

tatagcattt tgcttgctct cctttatctt gctatcactt gtagcacact ctgcacaatc 1260

cccacccacc ccagcatggt ggggcccctg gctcactctc cccacaagga atgtgcatgt 1320

gtgcaaaggg taggcctgga tgccatttct acaaacagtt gactaaagca gtccagagca 1380

gggatctttc tttgggtcat tgttatgtcc ttaattgacca gaataatgcc tggcacatag 1440

taggcactca ataaatactt gttgaatgga ggaatctgca aagctgctca gtcttcagac 1500

agcagcaagc agcaggaaga gaatcatgat gtaaccaaga gcaggtgctg gaggagctag 1560

ggagtgagtg gggatcttta aggacagcag gccctctgat ctgtctgagc gtgtgagcaa 1620

agcctagtta gggaggtgag ttctatctgc aatgagaggg ggcatgggtg gagacagata 1680

acaccaacac caacagacca gacctcccaa acccgaaagg agagacaaag tgtgattgat 1740

ttcccaaatg cagaaaagac tacccaagtc cccagccaag aagaatcgct ggcctgtgga 1800

tgaagacact ggctttgaaa tggctcaact acgtcccaag caccctttgt gcatcttaat 1860

tgccctggga aatcaggggag gggtggcagt ttggagcttg gaaagctgga aggtggggac 1920

gaaaggagtt tttgcccctc atcgccaatc cttgaggttc cctacagggaa cctcagacaa 1980

cagctgtctt agggggtttcc ttccagctcc aagcagctgt attatctata gctttgagag 2040

ggtctgtctc tgtcaatggg gctgattcct cagcacctgg cagggcctgg ggggactgc 2100

cagagtctgt tctgccgggc atgttagtag cacagactac tgggataacc tcagcaactg 2160

tataccttag cactgcagag ctgcggtgtt gtcagacaca ggcccttgca gttcgtggtc 2220

taccccccact tgcctgcacc ccccttatag ctgctcaatg tcctcagact gtaccttagt 2280

gccttggtta ccaagcaacc cccacagagg ccctcctcct ggattttccc ttccctgcta 2340

tgcccacccc accttcaaaa tgtacctgct gcacccatcc cccaggcctg cctcactcct 2400

tccaagctga cagctaatca gatggactaa tcgcatgcgc ggaagagctg tgctcgggaa 2460

gaagggggag atttcacacc aaatcgtgca gtggttgcct agacacggag ccagtcaagc 2520

ttgcatgccc cgttgctaga agagcatggc atcggttgtg ttctgtgctc gtaggctggg 2580

ccccagctcc ctttcctccc ctcctttgtg gctgttcctc aggcctgcag agggggaagat 2640

ttttcctgtc ctgttactct gcctcacttg ccagaacact agtcgcctcc agctctggat 2700

aaagctgagg ctgggtgggc cagacaacag ctgtcttagg ggtctccttc cagctccaag 2760

tagttgtatt atctatagct ctgagagggt ctgtctctgg gctcaggcct ttagtacctg 2820

tggggcagag ccaggaacgt gagtcaggca aaccgctcca aactaggacc aatgcagctg 2880

ggtcctgctc tctgcgcata gggggtcccc acatctttgg aatcccagcc cacctttcca 2940

ggctaccctc cacaggccgg gctccactcc catctgctga ggtttcccac cttgaatcct 3000

gctgccccga ttagctgtat aaccttaaag aatgcactgt ccctctccat taaatgaagt 3060

gcttggatgg attgctaaag gcctgtctgg ctcggaggct tggtgcctca aacacattgcc 3120

tgctggtcca aggaaatcag tgcctgagcc agagtcccca tctctaagct ccatggttat 3180

tgttcttgcc acctggctag gaaatgtcct tccagctgcc ccagtctagc tgcctcaccc 3240

tggggccatg ccccaactct gtcctaccct tctctgctgc tgacactcag ccccttccca 3300

gcttccagtt ggatacagga cctgggccag gagagcaggg aggacactgt ggaaatgcgg 3360

ccaggccatc aggggcctcg cagcaggggga ctggaggggg agcagtgtcc aggggccagaa 3420

gtgccctgcg ggagagccag gacattggct gcctgtggtc ttggtggtcg tggtcagttc 3480

cctctcctgc cagctgtgga atgtgaggcc tggcctggga gatatttttg ctgcactttg 3540

agccaccccg ccccctggaa ctcagaccct gcacagtcca tgccataaca atgacgacca 3600

cttccaattg tttcctagct ggagaggcgg ggaggggagc actgtttggg aaggggggga 3660

gcctggggga aatgcttcta gtgacaacag ccctttctaa atccggctag ggactgggtg 3720

ccgttggggg tgggggtgcc ctgctgcccc atatatacag cccctgagac caggtctggc 3780

tccacagctc tgtcctgctc tgtgtctttc cctgctgctc tcaggtagga gcgggagctg 3840

gaggctttac tctgggataa gggggctcca ggcttaggaa gggattcctc ttggaatagc 3900

aagcttcatg caggacttca tgcagagtac caggtccagt cactgggcac acatgtgcag 3960

gtctaaacat gggcgtatgt gccacaggag ttccataggg gcagatatct gcatctgagc 4020

atatgggacc aatatgcatt acagggtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgga 4080

caagcctgca tagtctgaat gtgagaaagg gtatgtacct cactgactga tacagaaata 4140

tcaagtggga gatgtgggtg tatgatcacg tctgtgaacg aggaggttcc tttgcaggga 4200

gggagaaaga atggtggaga aggattctag gaagtgtgta tgccttcatg tgattctgtg 4260

aggcctggaa tgtgtagcta cccctgtcca tgcatgcaga ggttcttgtc tatgcctcca 4320

cgagtaatga ctgttatagt atctgtaccc tctgcagaag gaactcacag gcacgggggt 4380

gtaggatgtt ccaggtcttt gtgtactgaa gaacacgcag atgtgatcta cttacgtggc 4440

acacatcctc aaatgtgcac atttgctgtt gaggcatata tggcaccctg gggttggggt 4500

gcggattgag gtatgggaag gaagcctgtc ctcatgtgag aggcatgccc aagcagcagg 4560

catgagtgtt catcagaggg cttgtggagg gtgtttggct atgtgatggg tgtgtctgtg 4620

agaggaagcc cacagtgtct tctctctgag aatgtccaca aagtctgaca acaaaatggg 4680

gacaggatgg aaaagaagtc tctttagagt catcagactt gcattcaagt cttggctctc 4740

ccacttgttg tcatgggctc ttggccaagc aagccatta acctcgattt cctcatctgt 4800

caaatggggt gaagactgag taggagaaag tgcattgaag tgcttttaga ctggaaagtg 4860

ccgaacaaat gtgaccatta gttgtctctc agatggctca aagaaaggta ccactagatg 4920

ggtacaggca ggggtgggac catctcgcag gagtgaagag gattagggtt caggttgagt 4980

tgacacagga agctgttaga actagttctt gaggaaaaag ggaatagggt ttgaaaaaca 5040

agaaggatgg gttgggccag agaagctgac caccctttcc accttgtttt ggaagagggt 5100

ggatgctgaa gaacagagcc aggagaagc aggaaggtgg gactggtcag gtgggcacag 5160

cctgccctga cacagcctct tccctctctc caggtcccct gcaggccttg gcccctttcc 5220

tcatctgtag acacacttga gtagcccagg taagaaaagc tgaagctaga gcgttgaaaa 5280

tctagtaaga ctgggcatta gagccccaaa gccagcctat ggaactctag tttctgccat 5340

gtgctggaga gaatatcttg gctatcaaga gctacttacc tgtgaccagg gggtccaggg 5400

atagatgagg gtcagagatg gaaatagtta gatttctgct cctacagggg acctgcaggg 5460

agctcagaat gctcttcctt cccctaggag agccccagct tgaacaatag ttagttgttc 5520

cttttaccct ggcaggtttg ctaggctgct cctctggtta aggggtaacc tcaaaaggaa 5580

gggactcact ggtaactcct cttgactctt gagcatggtg ctaggttttg gggctcccac 5640

tgaaggggag agcccaggga gggaagggaa gaatgggcag atgggagggc agccagcttc 5700

tgctcactcc aggcacagcc atgggagatt cggagatggc agtctttggg gctgccgccc 5760

cctacctgcg caagtcagag aaggagcggc tagaagcgca gaccaggcct tttgacctca 5820

agaaggatgt cttcgtgcct gatgacaaac aggagtttgt caaggccaag atcgtgtctc 5880

gagagggtgg caaagtcact gccgagactg agtatggcaa ggtgggtgtc aggctgatgt 5940

gagagtccac cctggccacc tgtacacctg ggtggacaga gaggggtaca cccatgcccc 6000

atgcccatgc agacctgagg atggtcccat tctccttccc tttgggaaga atccagaccc 6060

tccaaagagc cctctccttc ccaatatccc ttctgtcttc cctgtgagat cctggttcct 6120

tctctcttga gcactattgc cctgtcactc accaactcct aaccctcttg aggaaggagg 6180

gaaagcccag gctgacagga gggcttgggt gggggctctt gcagacagtg accgtgaagg 6240

aggacccaggt gatgcagcag aacccaccca agttcgacaa aatcgaggac atggccatgc 6300

tgaccttcct gcatgagccc gcggtgctct acaacctcaa ggatcgctac ggctcctgga 6360

tgatctacgt gagtgctgca cctggcccta cgttgggatc tctgttcttg ctccatccat 6420

gtccacccca ggagccaaga gtgtcttctg tttgtgtcta ggcagggtta cactctaacc 6480

tcgtcccaac atccttggtt caattccaac actctgggga ctggcattac tcagattgag 6540

tgcatgcaga ggttttcctt tctcttcttt ctctcctggg atctttctct aactcccaaa 6600

atcaccagcc ctcccccttc gcaactggca agtcactgct ccttttctat ccccagacct 6660

actcgggcct cttctgtgtc accgtcaacc cttacaagtg gctgccggtg tacactcctg 6720

aggtggtggc tgcctaccgg ggcaagaaga ggagcgaggc cccgccccac atcttctcca 6780

tctccgacaa cgcctatcag tacatgctga caggtgagag gccctggaag gtcttcctga 6840

agggaactgg gataggccgg gagggagggg gagaaggaag ggagaagccc cacgagagca 6900

tcctgtgcag ctcctgacct ttcctcccca ccctctcccc acagacagag aaaaccagtc 6960

catcctgatc acgtgagtgt agctgctact gtattccctt tcagaatctc cctgatccca 7020

gcctccagcc ccatcaggtg tcccaggccc cgaggcatag actcagcctc tttcccccca 7080

acctcgtgcc catctccttc ttctctgacc attaccctga cccctctctt tgctctccct 7140

cttcctttct gctttccatc cccttcatgt ttttcctctc ccacctcctt cacccactct 7200

ccaactcatc acgcgtttat ccagaagctc attatcgcca tgtcttcacc tctgcatttt 7260

ctttcccttc agaaacattt cccattcttc cagccctacc aacttttccc aaaacagccc 7320

aggcccatct cccctgttta ctttgcccag ccaagcacag gtcagcctgc ccaaaccagt 7380

gccttctggt ggcatccttc ctggtgccag ccctgacctc tgtgcatcag aagacagttg 7440

tggattttgg gtgtagataa cctggcagct tcttgcccag ggctcctact ctcgggtagg 7500

cccaggcatt ctctcctgat ttgaggcttg ctggtctcca gtagtattgt tcactgccca 7560

ataagcccct gtcttcacag cggagaatcc ggagcaggga agacagtcaa caccaagagg 7620

gtcatccagt actttgctgt tattgcagcc attggggacc gcagcaagaa ggaccagagc 7680

ccgggcaagg taggcctgct gccctccaag gtcctgtacc gcagaaggga gggagaagag 7740

ctctcacctg cctccttctt ggcctctgca gggcaccctg gaggaccaga tcatccaggc 7800

caaccctgct ctggaggcct ttggcaatgc caagaccgtc cggaacgaca actcctcccg 7860

cttcgtgagt ggtccctgac cttggccttg ggacttggac tggtggagga atggtctcag 7920

atatgagcct tcccccaact catcaccact ctcttccatc tctccagggg aaattcattc 7980

gaattcattt tggggcaaca ggaaagttgg catctgcaga catagagacc tgtgagtgcc 8040

atgaatctgc taggctcagc ctaagctcac ccttgctcta gaccatctgg tcttgacctc 8100

tctctctctc ccctccctcc ctctgttttt ctcctcttta agtctctgtc tgtaggtgtc 8160

tctgtcttca ggtctacata tctgtctctc tctgagactt cctctgcatc tttctccatt 8220

tctgtctctg catggctagg tgtctttctc tgggattctct ctctgagact atttctctcc 8280

ttctgggtct ctgtttccat ctctctgtgt gatctctttg tgtctgtcca actagtctct 8340

ctggctcttc ccttccctct gccttttgct tgctacattt atcattaatt ttccttgtgc 8400

ccaaacccta acttttcttt ctctccttct tctccccacc tgttcacaga tcttctggaa 8460

aaatccagag ttaattttcca gctgaaagca gagagagatt atcacatttt ctaccaaatc 8520

ctgtctaaca aaaagcctga gctgctgggt gagtcagagc caccgactga gaccaactat 8580

ctccatggca acctggtccc ctctgcttgt ggctggactc agctggcatg ccctgggttt 8640

catggcactg ctggattcaa ttcagctggg atgtggggcc aagccaagcc ctgtcctgtg 8700

ctgcttcctc aggccatgtg ctgtggcgag cagcctccat gagagccggg ggcttgtgtc 8760

ccaccttaac catgtttttt cccctagaca tgctgctgat caccaacaac ccctacgatt 8820

atgcattcat ctcccaagga gagaccaccg tggcctccat tgatgacgct gaggagctca 8880

tggccactga tgtgagtgtg tgaggaccca gccagggggt gggaggattg gcagtgaggg 8940

gcaaacaggg gtccaaaagc agagctaaga ctctggccat tggttgttta aagtggtatg 9000

gacctcctga gagccagaat gtgccctgtg atcagggtgt actggggaat ggatgaggtg 9060

agacaggact ggccagtatc tggtgttcta tcaaaccagg aagagctgaa agccatgaga 9120

cagagagaca cacaagccaa gaaacaagca tcaccgaggg agcaggacga ggagggaagg 9180

ccatgcagac ggcatggggc tggaggggct gttaggtgaa cagatgcaga caggtatgga 9240

aggccaggca ggaagcaagt gtagggccag gaagcataag tgggtaactc agaaaagctg 9300

tggccactga cactggggct gcggccaact gacgtctgct ctgctcttgc atcttacatc 9360

ctttgttttg gagttccctg catcttacat cctttgtttt ggagctcccc actgggtatg 9420

ggagtctcag aacccacagg gattaaggag acaagttttc tctccaactt acaagggatc 9480

tcacttaccc atcatacttc tttttctggg gtccgccaat atggggcctc cctacagaac 9540

gcttttgatg tgctgggctt cacttcagag gagaaaaact ccatgtataa gctgacaggc 9600

gccatcatgc actttggaaa catgaagttc aagctgaagc agcgggagga gcaggcggag 9660

ccagacggca ctgaaggtgg gaggcaggat tcttggggca gctgtaagtc atgaggcagt 9720

ctgctcagca gtatctctct tctcttttct tttttttttt ttttttgaga tggagtcttg 9780

ctctgttgcc caggctggag cgcagtggca cgatcttggc tcactgccac ctccacatcc 9840

tgggttcaag tgattctcct gcctcagcct cccgagtagc tggaattaca ggcatgcacc 9900

accatgcctg gctaattttt gtatttttag tagagatggg gtttcatcat gttggccagg 9960

ttggtctcaa actcctgacc tcaagtgatc cgcctgcctt ggcctcccaa agtgctggga 10020

ttacaggcat gaaccaccac acctggccag cagtcatctc tttaccaact ttgctacttg 10080

ccttttcctt ccagaggctg acaagtctgc ctacctcatg gggctgaact cagccgacct 10140

gctcaagggg ctgtgccacc ctcgggtgaa agtgggcaat gagtacgtca ccaaggggca 10200

gaatgtccag caggtgggtc catcttcaga tgataatggg tgggcagggt agggagactg 10260

gcatggtggg atgagagttt tcaagttcac tcttcccaac aaccctgctc aatatgggtc 10320

tctcctccac cttgcaggtg atatatgcca ctggggcact ggccaaggca gtgtatgaga 10380

ggatgttcaa ctggatggtg acgcgcatca atgccaccct ggagaccaag cagccacgcc 10440

agtacttcat aggagtcctg gacatcgctg gcttcgagat cttcgatgtg agttgggacc 10500

cctgggagtg ggagaacaat cactcactcg ctcccacatt caacagctat ttgcttagag 10560

ccagctgtgg accagacatg ggaaggcagt ggggactgtg tggtgacaga ggcagtcatt 10620

ttctctgtct tcaggggaag ccctccttca ctgccttgac atggagggga ccagccacgc 10680

cctgctgggc tcaggcacag tggacgggca cagccccaat ggccactcac accactttc 10740

tgactgctcc cacccctcat gccccctgca gttcaacagc tttgagcagc tctgcatcaa 10800

cttcaccaac gagaagctgc agcagttctt caaccaccac atgtttgtgc tggagcagga 10860

ggagtacaag aaggagggca tcgagtggac attcattgac tttggcatgg acctgcaggc 10920

ctgcattgac ctcatcgaga aggtgcctct ttggccttac cacctgaatt ctccctgcac 10980

acccaacaag aacaccatag acaaaatagc agcccctctc ccttctgggg aatataaaaa 11040

tagaaggggc tgaaggatcc gagcccttgg ttccagagcc tatgttgtgc tgagcaccca 11100

gcacagggtg gcaccaggga cagctgattt cccaggggct tgaaagtgga cacatggagg 11160

atgggcacct gcatgatgac ctcccacacc tgcatgttta ttgggcctgg tttatgcatc 11220

caaggatcag gaagtgtgag ggtggtgtag gggatcataa gagtgcacct attttcaacc 11280

ctagcatctc aggcatctgg gtcgtggagt ggtgtgtaca gcatcgatag aatccatatt 11340

cccagacttt cacaaagtcc ttctgtcatc agacaaaatn ctccaccttg aaccggttcc 11400

agtcagtaga taactgtact cagagctgag cctactacct taacacccaa catggcacct 11460

ccacgagcaa gtatattgac catagagcag aatccatgtc cacctgtgtg aaggacactc 11520

agtgatgctc tctcctgctt cctcagccca tgggcatcat gtccatcctg gaagaggagt 11580

gcatgttccc caaggccacc gacatgacct tcaaggccaa gctgtttgac aaccacctgg 11640

gcaaatccgc caacttccag aagccacgca atatcaaggg gaagcctgaa gcccacttct 11700

ccctgatcca ctatgccggc atcgtggact acaacatcat tggctggctg cagaagaaca 11760

aggatcctct caatgagact gtcgtgggct tgtatcagaa gtcttccctc aagctgctca 11820

gcaccctgtt tgccaactat gctggggctg atgcgcgtaa gtagggactg aggctcccgg 11880

tacagaggag cagggattct gccaaggttc tgagccaggt caattgctac accccaccac 11940

ttcaatgtca ggttgtaccc caaggtttac agactcagtg ggatggaact gggtgaagaa 12000

actgaggcag ccacattgaa ggcctaccag gggcaaatgc cagcaaggat gtaaagaggg 12060

gctgtgatct tactcacacc ctacctcccc acactgatgc ttcttttgtt gactctcctt 12120

cctgcagcta ttgagaaggg caaaggcaag gccaagaaag gctcgtcctt tcagactgtg 12180

tcagctctgc acagggtgag tgggacacag ccccagccaa cttggctccc catctgccca 12240

accccaccca gccccaccct tccctgcctt gttcatcccc tactcctccc gttccctgtc 12300

tccttggtgc attcgggacc attttcactc tgtcttctct tcccgtcatc tcctggcctc 12360

ttcacttatt tacttcccat ctcccttctc tcctctttcc cttctgtctc ccaccctctc 12420

tccttatccc tgtctccccc agggcccctt catctctgtg acttctcgaa ttctctccat 12480

ctctcttttc cttccttctt ctcctctctt cttcctgcat ctctttctgg cattttcttt 12540

acttttctct attgcatttt tggccacagg aaaatctgaa caagctgatg accaacttgc 12600

gctccaccca tccccacttt gtacgttgta tcatccctaa tgagacaaag tctccaggtg 12660

aggccacaaa ctagggccaa cccactgctg ggcatacacc gccctgggaa caggacctcc 12720

taggacatct ccctacctac caccacagtg gtttccaaac cacattcttc tcctcccatc 12780

ccgcatgagg cctcccaggg tacctccctc tcagcgctac cttgcacttg cagtgtgtaa 12840

tgagggtgat gtgtaaggca ctctgatgga cattacctca tcagtggccc tacaaccccc 12900

tggatagctg gttgtccaag acagaatctt agttcaactt taaatttacc tgcagaacca 12960

cttaagcccc caatgagaca tattcaaatt gttaatccat ttttggctaa ttaggagaca 13020

gaggctcagg gacctgggtt caagacctgg ctctgttctt gatcagctta gtgaacttgg 13080

cctgtcccgt aacccctctg ggcttaattt actcatctag aacattggat atatctgttc 13140

tggctaccac ctatgataat aggtgtgaat catataaaaa aggaatgtga aagcctttta 13200

gttaaaaatg tggctagaag aaaatgaaac aaatgattat tactaactga cttgctaaga 13260

ttacaagcta atcagtgaca aagccaggat cagaacccag aacttcagtc cagtgttctc 13320

acagactcct cctacttcct tcttgccaca ggcgtgatgg acaaccccct ggtcatgcac 13380

cagctgcgct gcaatggtgt gctggagggc atccgcatct gcaggaaagg cttccccaac 13440

cgcatcctct acggggactt ccggcagagg tgggtatgag ggtgcaccag agctcataga 13500

acagggggag ccaggctgcc ctgatgggaa tgggatctgc aggtgaccct agaattctat 13560

gggcagagca gatcactgca gagcatgggt gactctggac acttccctcc tcaggtatcg 13620

catcctgaac ccagcggcca tccctgaggg acagttcatt gatagcagga aggggggcaga 13680

gaagctgctc agctccctgg acattgatca caaccagtac aagtttggcc acaccaaggt 13740

gaggaaagga gactaattaa ttaaaggaag acatctcttt tccattgact cctctgatgc 13800

ttttcctgtt gtaatcatct tagcaaaatc tcttacctgt atgctacccc tcccagtgga 13860

acatctagca ccactcccct catcccagct ccagctgcca ttgacctcct ccctgcaatc 13920

cttttctagg ctgttaccct tcctaaggta atccccacca tctctttccc tcgtacccct 13980

ccctagtcat ggccaacaca caccttgcct gcaggtgttc ttcaaggccg ggctgctggg 14040

gctgctggag gaaatgaggg acgagaggct gagccgcatc atcacgcgta tccaggccca 14100

gtcccgaggt gtgctcgcca gaatggagta caaaaagctg ctggaacgta ggtgagagat 14160

ctcaagagga ggtttcccgc ttctctgagg cccaggctgg ttcaggggca gtgtcaggaa 14220

aaaaagctca gcagatcttc aaacacagag acctgcagga ggggctcata tgaacacact 14280

gcagtcacag ggtcagaggc ctcaggaagg gtgggaggga tgaaggaaat gggatattcc 14340

caaggtttca ggacctcagg taggaaggag gcagaggctc agcactcctt tcaatgggcc 14400

cctgcagaga ctccctgctg gtaatccagt ggaacattcg ggccttcatg ggggtcaaga 14460

attggccctg gatgaagctc tacttcaaga tcaagccgct gctgaagagt gcagaaagag 14520

agaaggagat ggcctccatg aaggaggagt tcacacgcct caaagaggcg ctagagaagt 14580

ccgaggctcg ccgcaaggag ctggaggaga agatggtgtc cctgctgcag gagaagaatg 14640

acctgcagct ccaagtgcag gcggtgaggc tcctgggcta ctgttggctc ttccaccctg 14700

ctctgccttc acttcccaca accctgcacc tccctgcaca ggggctcact gtcttgttcc 14760

ttcagtcaga ggactctggg atcaacactt ctaaagacct ccaaagaggt ccctcaggag 14820

gaggaaaggg cagagggaaa agagctgact tttaaaagca caggctttct acttgggttc 14880

aacctagata cacagcttaa ttattttgtg accttgggca gaccatgcat catctttgag 14940

cctcaatttc ctctagacag ggaataaagc tgtccatttc attaagccat catacataag 15000

tgcttggcat cagcaaatgc aattcccacc cttcgatgga ggagaccaga ggaagaggtc 15060

cagatgaaga tttctggttc ctttcctctc tgctcccctc cccagtgttc ccaagttata 15120

ctatcctgaa actcttcccc tccccatact tctgaagctc ttgccaagtg gggatatcct 15180

agagttaccc tcctatttga gtgatgtgcc tctccttccc tctacctgca agaatgagga 15240

ccttaccccc tgaacagcct cccctctgtt cctcaccttc aggaacaaga caacctggca 15300

gatgctgagg agcgctgtga tcagctgatc aaaaacaaga ttcagctgga ggctaaggtg 15360

aaggagatga acgagaggct ggaggatgag gaggagatga atgctgagct cactgccaag 15420

aagcgcaagt tggaagatga gtgctcagag ctcaaaaggg acatcgatga tctggagctg 15480

acactggcca aagtggagaa ggagaaacac gcaacagaga acaaggtaag ggcagctccc 15540

tttggctcca gcccgggtct catcaggact ctcagaccat actgaccttg acccaggcta 15600

gccactcgca tccagagagc agcatggacc ttgatcatgg agctccccac agatggcacc 15660

aagctggtga cctttgaccc taaaggagat gggattcttg gtcggcaggt gaaaaacctg 15720

acagaggaga tggctgggct ggatgagatc attgccaagc tgaccaagga gaagaaagct 15780

ctgcaagagg cccaccaaca ggctctggat gaccttcagg ccgaggagga caaggtcaac 15840

accctgacta aggccaaagt caagctggag cagcaagtgg atgatgtgag tagattgaga 15900

gttgtggggc ctagatatgc catgtctatc tgtgcccaga gctctgtgtg tgtgtgtgtg 15960

tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tttgtgttta tgtttagagg ggggtagggt 16020

tagaggatgt taacttttct gatatctttc tagaaaggag ctggggaatg aaaagtcaga 16080

ggtggtgatc aattctgaat tatgtatcta atctcactgg tatgatcaat aatagagatg 16140

agttttctta tttaatttaa gtgcaatccc atatatctct gcctttgaca caagatttag 16200

agggtcatct cataagaaca tgataaattt ctttggaaat tttgtgaggc tccccaagta 16260

gaatttttga aattccaata tttaaggcag aatctgatca ttgttccctc tgatgcacat 16320

atgtctgttg actcacttat cactctgtat tttaattatt taagcttcta tctcagccat 16380

aagactacaa gcttccttta gataggaact tagatctatt caatttgggg actgcagtgc 16440

ttagcaagag cctgatgccc agtaggtgtt tataggatgc tgaaagaatg aatacgcgcg 16500

gccagccggt ggctcacacc tgtaatccca gcactttggg aggccaaggc gggtggatca 16560

cgaggtcaag agatcgagac catcctggcc aatatggtga aaccccatct ctactaaaaa 16620

tacaaaaatt agctggatgt ggtggcgtgc gcctgtagtc ccagctactc aggaggctga 16680

ggcagaataa tcccttgaac cagggaggcg gaggttgcag tgagccgaga tcgcgccact 16740

gcactccagc ctggcgacag agtgagactc cataaaaaaa aaaaaaaaaa aaaggaggaa 16800

ggaaggaagg aaggagagaa agaaaggaga gaaagaaaga aagaaagaaa gggaaagaaa 16860

gaaagaatac ggctttgggg tttcatccca cagtcattta ttcctttatc tataaaatat 16920

tgattcagca tcccctatat ctaggcaatc tcacagtccc ctaataacat ttggcccact 16980

ctggggaagg tttcccaagt cctgaacaca aagatttacc aagtcctgag gtaactgaac 17040

aacaaaatca ccaagtcagg atgctttgcc tcactgggcc tggctccttc tctctaccag 17100

ctggaaggat ccctggagca agagaagaag gtgcgcatgg acctggagcg agcgaagcgg 17160

aagctggagg gcgacctgaa gctgacccag gagagcatca tggacctgga gaatgacaag 17220

cagcagctgg atgagcggct gaaaaagtac tgtgtcccct ccctgctgcc cttcctcctc 17280

cccagcccat aacagtacaa gcagacccaa gacagccatc ctctgaggca acaacctcac 17340

gcagcctcca caagtggagg cgcaaaccat gggaaaaggc ctcccaggca agggctttcc 17400

tcccctagtc ccagatatca gggttagtta gatggtcctc cctggcaaaa agtccattgt 17460

atcccatgag acctttcttc ttgttaggca ctcagtggga atgaaaataa cagccttccc 17520

ccttcataca catctctctt tggagaccag ttccttgaca tggaggaagg ctccacatga 17580

gagcctctta ggaaagtccc tttgtggctt ttcacagaga gcccacagggg caccaagaaa 17640

tgccctcaca ttaaagaagt caggggatca gggccatttc tttgttttgc ttaacttgtg 17700

gttccaattc ttagtgtaag ttcttccaaa atgtatagac tttccagact tggtaaacag 17760

tgagcttgca gaacatattt tgataattga aggtcttatg gtgtttcagg ctcaggactt 17820

tgaaaggctt tgagaatcat cataatctac atcctcatca agtttgtgtt tgcacaggaa 17880

tcttcatacc ttcccaggca cagaatgtgt tgaataaatg aataaataca cgatcctatt 17940

tgattctcaa aacacctttt aaggatggta tctttaattt cactttacca aagaggaaaa 18000

taagtttcag aaaggttagc tgagttaccc aaagcaatag tgacaaaatt aaagaccatc 18060

agagctaaaa aggacctttg aaacaatcta gtccgacccc ctcattttac attatgagaa 18120

aacaaggacc aaagatggga atgtcctgta gagcccaggc cccatcaccc tgtctacagc 18180

actcctccca ttacattctt gttggatttc aatctaccgc aggtgttaca cttccaggga 18240

tcctacatca cccccagaag ctgtttttct aatgaaatcc tactctttac ctgtatcatt 18300

accattttca accactggtg gaccctcctg aggccccacg agtctccctt acctcaccat 18360

ccctccttcc ccacagaaaa gactttgagc tgaatgctct caacgcaagg attgaggatg 18420

aacaggccct cggcagccag ctgcagaaga agctcaagga gcttcaggtg aggtctggac 18480

accacgcgg gatgctgagt ccctgggctg gttctgtttc ccctgccccc tcctctaggg 18540

tgtcctgttc acagcattac aggaactgat tatgtgtcca tggaggcagg cacgcagcta 18600

tcacagtgtg cccatggata catccaaaag gggtcaggtg atccacagag accctgaaaa 18660

ggtgcacagg cctggagggc agaattcctg gcttcctgtc tcagctctgc cttagttggg 18720

tgacctctgg gagccccactc cagatctctg ggtctccatg ccctcccctg ttacatggag 18780

gtttttaaga gtccttcgac tctgatggca tgactttcct ccttactcta gagcagaagc 18840

tgtgttaggt ctttagttgt ctggtcagac aaatacggct ctctcatttt caacttttgt 18900

tattttttta tgactaaact gcttaaaata tccccagaac acaggtctct ttgtccatct 18960

ttaagagcaa gtttcaacca ctttccttcc aataaagagc catgtagact gtcaggccct 19020

cgcctaaagg aaggctcttc aaactatctt ccccaaatcc tttaattccc acctctccag 19080

tggaagagct aaactgactt gctgttccag agaagccgag agccttttag agccgggggg 19140

attccagtgg agggggtccag gcggtgggtc tgagcccttt gtgtctgacc aggcacgcat 19200

cgaggagctg gaggaggagc tggagtccga gcgcaccgcc agggctaagg tggagaagct 19260

gcgctcagac ctgtctcggg agctggagga gatcagcgag cggctggaag aggccggcgg 19320

ggccacgtcc gtgcagatcg agatgaacaa gaagcgcgag gccgagttcc agaagatgcg 19380

gcgggacctg gaggaggcca cgctgcagca cgaggccact gccgcggccc tgcgcaagaa 19440

gcacgccgac agcgtggccg agctgggcga gcagatcgac aacctgcagc gggtgaagca 19500

gaagctggag aaggagaaga gcgagttcaa gctggagctg gatgacgtca cctccaacat 19560

ggagcagatc atcaaggcca aggcaggctc tgctcggcct cccctcctcc aacttcctcc 19620

tcccacctcc cctttctgtc catgtagtgt ctcttccttc atggttcatt ttcccacttc 19680

ccttcctctg cctggttctc agatcccctc cttcttttca caccttccct cttcttttct 19740

ccttcagttg cacctcttac accccttcat tccccccgcc cccaacatcc atcatataac 19800

tctccttccc ttctcaggct aacctggaga agatgtgccg gaccttggaa gaccagatga 19860

atgagcaccg gagcaaggcg gaggagaccc agcgttctgt caacgacctc accagccagc 19920

gggccaagtt gcaaaccgag aatggtgagc ctagagcagg ggctccatgg ttcccaccac 19980

agtctcccca acctcctgag ccactccagc ctcgggcaca agccaataca cgcacacaca 20040

gacacagagt ctctccagaa aatggggcct gagggctaga ggaggaggtg gggatagaga 20100

ggagtgctga tctagaatgg tgcttcccca ggtgagctgt cccggcagct ggatgagaag 20160

gaggcactga tctcccagct gacccgaggc aagctcacct aacccagca gctggaggac 20220

ctcaagaggc agctggagga ggaggttaag gtaagggcta gactcgggcc acctggccca 20280

agcaaggagc acactgacta gccttgcatc aaatcacttc ccttcccagc aataaggctg 20340

gctgtggacc aacagttctc caagaattct aaaaagaaat cacagcccaa accctgccac 20400

atgccatcta ctccccaggc tgaggccctc tgcttctgct gcatccctgg acgggaggca 20460

gggtgtgaga aggaggagag aaaatgaact agggaggcac gagatggcct tgcccgggca 20520

cggctagggg atgtagggac tgcacaaatg ggtcctggat tgctagggcc accctgaaaa 20580

ctggagaaag aaattcaact ggagcctatt cctagcactt tgctggactc atgaaaaagg 20640

agaaagaaca tgatttttaa aaaactccag ggttcatact aacctgcctc ttatagctca 20700

gtgatcttag gttagggaag tcccctaacc tttctgagcc tgtttccctc tccccagcca 20760

ggtaaaatgg cggtgatgat gaccatgtct ggtgtggggga ggagctcagg gccaatggcc 20820

atgaagcttt tcttaaatag ttcactgctg tattggcatt ctttgttatc agcaaacagt 20880

aaaccatgac aggtccactg ggggtttgga catagatgag gccaagggat gatgttgaga 20940

ttggggtgag gagaagggca agggtggggt tgctttatgg agaaagctga accccacctcc 21000

tggtgcccac ccctccccag gcgaagaacg ccctggccca cgcactgcag tcggcccggc 21060

atgactgcga cctgctgcgg gagcagtacg aggaggagac ggaggccaag gccgagctgc 21120

agcgcgtcct ttccaaggcc aactcggagg tggcccagtg gaggaccaag tatgagacgg 21180

acgccattca gcggactgag gagctcgagg aggccaagtg agttctgagc agcctgactt 21240

ctggctgagg cccctttgca ggcaggactc agcccagccc cagcctcagc agatcccaca 21300

caggcgatgc ttagctagtg tttgacaaca caggaggact ctgccccggc cccacctcct 21360

tctcctctca gggaagcttt ttgctcgaat tatgtttctg atccgaatat aagacgaaca 21420

aaaggtttgt ctgagggcag agtgcttccg tctggggcag ggcactgtgg cagggaaagg 21480

cagtggggag ggctgcagaa gcccatacct cctcaatgtc catagcgcag aggctgggcc 21540

agggtcagag ggtgcctggg tctccacgcc ctgctgggat cctctgcctg atgttctcgg 21600

cccctgggac ctgtcctcag gcttctccag ctacacttct gaggtttcaa ggattgtctt 21660

tgagaaggtt catgttgttt acctcttgtc cccatccaca ccctccatcc tccccaccct 21720

ctgccacctc ccctgggcag gaagaagctg gcccagcggc tgcaggaagc tgaggaggcc 21780

gtggaggctg ttaatgccaa gtgctcctcg ctggagaaga ccaagcaccg gctacagaat 21840

gagatcgagg acttgatggt ggacgtagag cgctccaatg ctgctgctgc agccctggac 21900

aagaagcaga ggaacttcga caaggtgggc cctgggtggg ggccgcagcc agcatgcagg 21960

gcaagggggc atgaggggtt cagtgagagg ccagagccat cctccttgga ggtgggggag 22020

gagggctgag cccaggcagg tcctgagaca gaccctggac atggggctga ggctgggggg 22080

ctgaagagtg agccttgtcc ccgggcagat cctggccgag tggaagcaga agtatgagga 22140

gtcgcagtcg gagctggagt cctcgcagaa ggaggctcgc tccctcagca cagagctctt 22200

caaactcaag aacgcctatg aggagtccct ggaacatctg gagaccttca agcgggagaa 22260

caaaaacctg cagggtgtgc tggggggccca agaggctggg gaggggctgc actggcagtg 22320

ttcccatatg gtgccacca agggctccaa gaggcgtctg ggcaaagagg cgtgtccctc 22380

caactccact ggacctcagc agccctcaac cgagttaccg tgttccccac acaggaga 22440

tctccgactt gactgagcag ttgggttcca gcggaaagac tatccatgag ctggagaagg 22500

tccgaaagca gctggaggcc gagaagatgg agctgcagtc agccctggag gaggccgagg 22560

tgtgtgtgtg tgcagggcac ggggtgcagg gagctgagct ccaggatttt ggctccctgt 22620

tctcatcccc cgctttgtcg ttttgttcac tgtcccatcc ccagagccta ggacagagcc 22680

tggcacatag taggcattca gtgaagattt gtggagtgaa tgaatgacca gtcactgaac 22740

tctcttcaca taggcatttt acccartgca tcttagctga gcttcccatt tccacatacc 22800

tcccggttcc cacctctgcc tccctctgtt ttcttttata ctctatacct gattgcctct 22860

gtttccttgg caacactttt ttcttcattt tcctccttat cttcatagct ggctctcccc 22920

tgtgagggcc tgaatcactt tctcttaggc ttatttttat tttctcttag attctctttt 22980

cctccacctt ctgcttcttt gaaccactta caccactctt gaagtcactt cgtatccatg 23040

attagtgagc aggcccccac cctgccctgt gccctgactg tctgcctgca tcccctcccc 23100

caaccccttc ccaggcctcc ctggagcacg aggagggcaa gatcctccgg gcccagctgg 23160

agttcaacca gatcaaggca gagatcgagc ggaagctggc agagaaggac gaggagatgg 23220

aacaggccaa gcgcaaccac ctgcgggtgg tggactcgct gcagacctcc ctggacgcag 23280

agacacgcag ccgcaacgag gccctgaggg tgaagaagaa gatggaagga gacctcaatg 23340

agatggagat ccagctcagc cacgccaacc gcatggccgc cgaggcccag aagcaagtca 23400

agagcctcca gagcttgttg aaggtactca cccagagggg actggcctcc acgtggcctg 23460

gcgaagcagt agtgtcttga tacaggcacc agattcctcc tgcccctagg ttactgcagg 23520

gacctctgac aggtgccttt agtgaaggga accgaggctg gctccctgct catgcccact 23580

ctcctgatcc tcaggacaccc cagattcagc tggacgatgc agtccgtgcc aacgacgacc 23640

tgaaggagaa catcgccatc gtggagcggc gcaacaacct gctgcaggct gagctggagg 23700

agttgcgtgc cgtggtggag cagacagagc ggtcccggaa gctggcggag caggagctga 23760

ttgagactag tgagcgggtg cagctgctgc attcccaggt gagcagctcc cctgctcatt 23820

cctgaaggga gcacaggctg gggctcagca agcaaggctt gagagctatg catagatgct 23880

caatgctttt cctgctctgc ccaaccctcc cccaacccag aacaccagcc tcatcaacca 23940

gaagaagaag atggatgctg acctgtccca gctccagact gaagtggagg aggcagtgca 24000

ggagtgcagg aatgctgagg agaaggccaa gaaggccatc acggatgtaa gtcccccact 24060

ccaccgaccc gatccagacc agtgtctctc cgtgggctgg gcagcaagtg tgtgaggact 24120

tgaccagacc atgtgccacc tctctcctgc acacaggccg ccatgatggc agaggagctg 24180

aagaaggagc aggacaccag cgcccacctg gagcgcatga agaagaacat ggaacagacc 24240

attaaggacc tgcagcaccg gctggacgaa gccgagcaga tcgccctcaa gggcggcaag 24300

aagcagctgc agaagctgga agcgcgggtg cgggagctgg agaatgagct ggaggccgag 24360

cagaagcgca acgcagagtc ggtgaagggc atgaggaaga gcgagcggcg catcaaggag 24420

ctcacctacc aggtgcgacg ggcggtgact ccaggcagag ccctggcacc atagccacag 24480

tgacaaccag ctgaggagaa tgaagagttt gctcttagcc tcttccaggg cgaggatggg 24540

aatgcagccc ccgtttaaac tttgcctagc cctgccccac tctgaatgtc ccctagctca 24600

gaggtcagtc tctgagcctc ctggcctggg agctccatct caacacccac tcttagccca 24660

tgcgccaggg gacacacaca gaattccaga caaagctcac ccagtacagc tcaccagaaa 24720

gcaaatatgt agccagggtc accccaaaa gacaccaaaa acacaccat cccattcaaa 24780

caagcataaa aactttttca ttgcccaggg cagtgtggac tctagctttc tgaggtgttt 24840

tctagaatca gactctgaat tagaatttgt ttcttttcca atccaggatg tcactatctc 24900

tctaagcaca ctttgtcctt aacttagatt aagtcagtgt tttcctggcc tgcattgcag 24960

cccaggaaat tccatcacac tcatagcccc aaagtcactc ccagcatgtg ccgaggtcca 25020

gagcaggggga cttcatcccg catggggtcc ccggaccctg aaaaacctgc tccctggagc 25080

actgcacaca cacaattttg tgcataattt caggagtccc actagttctt tggactcttt 25140

cctggtacag atcatttaaa atatttacac atatttggta agaaattata aggagaattc 25200

aagtgtttag tgaggatcag aaagtagaat tgggtcagga tatcagatga agcagggcag 25260

ggcggagggg atgctacctt ctatgactgt gccatcttca ccccctgcct accctctggc 25320

ccccagacgg aggaggacag gaaaaacctg ctgcggctgc aggacctggt agacaagctg 25380

cagctaaagg tcaaggccta caagcgccag gccgaggagg cggtgagtga ccctgctggg 25440

gactaggccc aggggaggca taggagct cgtcccccaa gccaggaggtc tgagaaccca 25500

ggccccctct cacctcatgc tcccacctcc cgcaggagga gcaagccaac accaacctgt 25560

ccaagttccg caaggtgcag cacgagctgg atgaggcaga ggagcgggcg gacatcgccg 25620

agtcccaggt caacaagctg cgggccaaga gccgtgacat tggcacgaag gtgggtccct 25680

cttttgggct ttgctagtca cccccacagc aggcataccc agacagagca ccctcaaacc 25740

cgggatgctt ccttttcatt tattccaacac acttgagcca catggcccca gaggccaagg 25800

taatgcagtt ctctgtgatt tcaaggattc ttccagctct aacttttttt tttttttttt 25860

ttgagacggt gtctcactct gccgcccagg ctggagtgca gtggcacaat ctcagctcac 25920

tgcaacctct gcctcctggg ttctcctgcc tcagcctccc aagtagctgg gattataggc 25980

atgtgctacc atgtctagct aatttttata ttttagtaga gatgcggttt caccatgttg 26040

gccaggctgg tcttgaactc ctgaccttgt gatccactcg cctcagcctc ccaaagtacg 26100

ggattacagg cgtggagcac cactcccagc tcagctctaa gtttcagtgg ttctgggaat 26160

ttcagcgtta gggaggggtg ccacctagaa gaccctaatt tatatacaac cagtatatcc 26220

cagtttcccc ccaactgcta gtttccagtt cccttccacc taattttgtc ctgcaatcat 26280

tgtaactaaa acctaaaatg gacatcccag agaacccggg cctggctttg ttctcgccca 26340

ggcaacttcc ctctgaccaa ccccagaacc ctaccctcca gaacttgccc cccgaccatc 26400

gatccctgga aacacatgtg ctgctgccac aggggcttcc acctgcccac aaagcaggcc 26460

ccccaccatc agaccctct cacctttgtt cccatgccct gtccctgccc aataccatct 26520

ctccaaggac tgattggact ttgtttcctt tcaaaagggc ttgaatgagg agtagctttg 26580

ccacatcttg atctgctcag ccctggaggt gccagcaaag ccccatgctg gagcctgtgt 26640

aacagctcct tgggaggaag cagaataaag caattttcct tgaagccgag atcctgactc 26700

cagactcttc ttcactgccc tgaggacccg gggatggggt ctggcatagg ggaggtggaa 26760

atgcacggga gtgggggcaa attcaaaggc ctggcctagg aggccttcgt ggagcagcac 26820

tgaaaacccc tttcacttcc cccaaaccag ccaaatgcag tagaggcgac acctctcctc 26880

ccagccttct ccctggtcca gccaggattt aaaggagatt tagggagacc cctaaagttc 26940

acctagagca ccctgagccc atctactgtg gacctgcacc ccagcttccg tcaaccctgc 27000

cccacccacc accatggata ctgcccaggg ctgggcaaag ggcagcggtt cggggtgtgg 27060

gtgctggcca gtgccctttc tgagcccgga gaagttgtga accactgggc caggtgaggc 27120

aatggctgcc taagtgccag tcccaggcga gtctctgaga gtacacaagg attcacagct 27180

gtagcctgca tcccctttta aatagagcca ccaccctctc cagccaccgc cagaagccag 27240

cataccccct gccccaccaca ggtggagaaa cacaccctca gccccacctgc cgtcctcctc 27300

agctcctcct ggtgccgagc ccccttggcc cagggtgctc ccttcttccc actcatttct 27360

tcccattgta attcccatta ccaagccctt tgtcctctgg gcttgtggga agatggggtt 27420

ctcctggccc atttatggca agccattctt gaggggaggc aagacagatga ggggctgggg 27480

aaggttatgt gtgaacagga ctctggggag agggagggaa gggatggctc ccgagagggc 27540

caggaggtgg aggaaagagt aagtctcctc caactaaaca agttcagccc ctttggagtt 27600

cctgccacag ctggattgct taggcaggcc tggtactgtg ggccacagag gtcaggggct 27660

tgaaaggcac tggggctgtc ggaccccact gggaacctgg gaaggagtgg ttcaaccctg 27720

gactggatct cccccaaccc accaaccata cccactcgga gcccactcct gctctctacc 27780

cagctctgag ctctgaggac tgcggcagca gagccaaggg cctaagccgg agaaggagaa 27840

acactactgg acaatagcag gtgacagcag aggagaaaaa gggggtaggg agggggacaag 27900

gaaagaacct gagggggtgg cagtggaca agagattgag caaccacagg acaaggtaac 27960

cagaggaaac agagacaagg aaaaaagatg caggaaatga ggaggcccca gaaagatccg 28020

cagcgccgcc acctttctcc tcatagccac ctccattctc cttccctagg ccaacccaga 28080

cagagctcct caaaacaggg atgcttcctt ttcatttatc cagctcaatt gaaccacaca 28140

gccccagagg ccaaggtaat tcagtatctc tgtgatttct aggattcttt cagtcctttt 28200

tgtttttgtt tttgtttttg tgacggagtc tcactctgct gcccaggctg gagtgcagtg 28260

gcacaacctc ggcttgggcc gagatcccta gggccattcc tctggctagg aacagacttg 28320

gacagcaatg caagagataa ctagaaacaa aggagagaac ccttgaaaag ggagctatgc 28380

agagagaccc agggcaccta agcaaccaga gcaagagaga ccgcaagaga tacagatc 28438

<210>2

<211>20662

<212>DNA

<213> cMYBPC gene

<220>

<221>misc_feature

<222>(7506)..(7516)

<223> n is a, c, g, or t

<400>2

catggtgagt agcctggtgt gacgtctctc aggatgcctg agccggggaa gaagccaggt 60

agctttagga ctggggttgg gtctaagtgt gggagcatgg gggtgtctac aattggggag 120

caggctagga ggagttcttg agggggttga cgaggacgtg gcctctgagc cccaggacag 180

gggcagccag tcctccaggg tccttttttg attctcggtt tctttttccc atccctctga 240

tgagaggtga aggtaaaccc aaggtcaggg acgcagagag gggacacagc caagacagga 300

aagcacccgg ggtcaaccca ttctgagaat gcaaagggac cccttccctg tcccattgcc 360

ttcaccttcc acaagcaaggt tccccgcgga cagaacccac gtttagtatg gagctgagcc 420

aggctcacgg catgccttca gaccagtctc taccactat gggtctcagt ctcccaacct 480

ggggtgccgg gggtggtggt ggcattgggt tggttcctcc tggctgtgac attatctgct 540

tcatgagcct gtacggtgtg gcccccatgt tgagggtgac acagcccctc ttctctgaga 600

gaggacagtc aggagcccct gaacccaggc caagggcagg gaaaatcaca gcccagggac 660

ctgggcctgg gctgtgccct tatgaaggtt gtccagggca gagatccagg tcccatcccc 720

cagcgccctc ccaggccagc tttatctcag ttgctgatac ccagcctgag gtccttggag 780

gacaaatttg gccacattcc aagtaggcag acttccccgc cccctgccaa gggcccgaaa 840

tggttggggg aaagggagct gggttcatca cgggtgtctc caagcaggga aagtggacac 900

atgcagaaaa agtcttgttg ggaggtcagg accacagccc tggctctccc gactgctagc 960

tgtgtgatct tggaggagcc gcttagcttc tctgagcctc agtgtcctcc tctgagaaat 1020

gaacagaata actgcaacta cctcagagaa ctggtagagagt attacgtgag gggatgcata 1080

gaaagtgcta gcacagtatt tactgagagg ggctgcaggg gcggggtgca cgctccaacc 1140

aggggccgct gtagcctcca cctggccctt cagtctcagc ctttagcaag aagccacggt 1200

cagtggaagt ggccgcaggc agccctgccg tgttcgaggc cgagacagag cgggcaggag 1260

tgaaggtgcg ctggcagcgc ggaggcagtg acatcagcgc cagcaacaag tacggcctgg 1320

ccacagaggg cacacggcat acgctgacag tgcgggaagt gggccctgcc gaccagggat 1380

ccttacgcagt cattgctggc tcctccaagg tcaagttcga cctcaaggtc atagaggcag 1440

gtaagatcct gatcagcctt ccctgagttt gggctgctgg gggagggcag cccagcgact 1500

ctccatccat ccagggaaca ggaggtgctt tcacactttc ttgcctttgc tgtggctgtg 1560

ccccccgcca ggaacgccct ttcccccttt tctgcacatg acctctctga tacccccatg 1620

tgccatcctg gcttgttcca gccatggggt agtggggagag atgaacaaga cacaacccca 1680

ctctccccct gcagacctcc ctggggagat gacctgcaca gacggccggg aagccccagg 1740

agctgtggat gaggggagtg cagcccagtc tgaggggctg ctctaaggct ggggcagagc 1800

agggatggac ctggagctgt tcagggcaga tgcagaggac atgggaagtg atggcctggg 1860

acggggagga gaatgtgaga ggcaggctgg ctctccagct cagcatgagc tgcctctgag 1920

gggcccaggc agcgggagga cagccatggc agactttcct catccacagc gggctcatgg 1980

gtccaatgct caggagagag agggtggagg ggtctctggt tagtggctgc accacgactc 2040

ccatctgatc ccctccagag aaggcagagc ccatgctggc ccctgcccct gcccctgctg 2100

aggccactgg agcccctgga gaagccccgg ccccagccgc tgagctggga gaaagtgccc 2160

caagtcccaa aggtgagagg ggctgggcag ggaggggcag gatccgtggg tagggcgtgt 2220

ccagggcagg tctcaaaagc ctttgctggg ccaggtgcgg tggctcaggc ctataatccc 2280

cagtactttg ggaggccgag gtgggcggat catgaggtca ggagattgag accatcctgg 2340

ctaacatgat gaaaccccat ctctactaaa aatacaaaaa attagccggg cgtggtggcg 2400

ggcacctgcg gtcccagcta acttgggagg ctgaggcagg agaatggtgt gaacctggga 2460

ggcggagctt gcagtgagcc gagattgtgc ctctgcactc cagcctgggt gacagagcaa 2520

gactccatct caaacaaaca gaaaaagcct ttgctcacag ggtcaagctc agcagctctc 2580

aatggtccta cccctggagc ccccgatgac cccattggcc tcttcgtgat gcggccacag 2640

gatggcgagg tgaccgtggg tgagtgtgag ctgctgtgcc cagcattggg gtgggaaggg 2700

ggggcagcag gacactcccc aagccgggcc tgtcgccctg cctttgcagg tggcagcatc 2760

accttctcag cccgcgtggc cggcgccagc ctcctgaagc cgcctgtggt caagtggttc 2820

aagggcaaat gggtggacct gagcagcaag gtgggccagc acctgcagct gcacgacagc 2880

tacgaccgcg ccagcaaggt gggccagcac ctgcagctgc acgacagcta cgaccgcgcc 2940

agcaaggtgg gtccaccggg gtcgtggaga cacggagaga ggggacatgg agagccctag 3000

aaggcacaag gcacacagag ggcaccatga gacccagagg caacacacgag ctgcaagcaa 3060

ggggtgtggg gaatctggga ggcaggtgga ggggccaggg gcccacaggg agatcccgga 3120

ggcctggggg ccagggagtg catgaggctc atgggagcac tgtcggggac ctgggagact 3180

tggtggacat gatcacctca aggggggcacc taataaggtt tggccgggggg tggggggggcg 3240

cccagggtgg tcaggaagcc tggaggacac tgtgaagcat gggggcacca gctagtgggg 3300

ttgtaaagcc ccgggatggg tgagtatcag ggctaagggc tcaagggact ctcagggatt 3360

aaccagaaca attcctcatt atacagacac ggaaaccaaa gcttgagagg ttaagtggct 3420

gctccaaggt cacttagaca taggcctaga accttaagcc ttcgtttttt gtgttttgtt 3480

tttttccgcc ctgccccacca gaaccttaag gcttaagtcc ccttgtttgt tcactccaag 3540

cctctgcttt tcccaacctt ccccagtcct gggctaggca aggggggcctc tagcaagaga 3600

ggccaacatg ttctgctgtg cggggctgtg cattgcacaa aagcctgtat ttcaccacaa 3660

ccgtgtgcca gcagatggca atcatgggtt tcgagatagt cccctttctc tgtacaaaaa 3720

ctctgtatgg gttaactaaa gcctgagggt gcgaagatcg atgagcattt gtcactccca 3780

gcctccttta aatatatata tatatatata atttgagaca gggtctcgct ctgtcgctca 3840

ggctggagtg cagtggctca atctcggctc actgcaacct ccacctcctg ggttcaagcg 3900

attctcccac ctcagcctcc cgagtagctg aaactagagg tgtgcaccac catacttggc 3960

taattttcgt tatttttagt agagacggaa tttcaccacg ttggccaggc tggtctggag 4020

ctcctggtct tatgtgatcc gcccgcctca gcctcccaaa gtggggatta caggcctgag 4080

ccaccgcgcc cggccactcc cagtctcctt taagggtgcg gagccttgtc tcccggcccc 4140

tggtgtcccc tgacgccccg tccctccatg cacacaggtc tatctgttcg agctgcacat 4200

caccgatgcc cagcctgcct tcactggcag ctaccgctgt gaggtgtcca ccaaggacaa 4260

atttgaatgc tccaacttca atctcactgt ccacggtgag ggggccctgg tgtctgtcct 4320

gggctcgggc tccccatggg tcctggtctc ctacctcctt ttcccaacac taaggaggat 4380

gcctcgtccc atccagacat gagtgctggc cacgtgccca gtgctgcaca cacagggtgt 4440

gagagaaacc ccaaggcttg cagggtaggc gtgggggctt agggctgaat ccgggtctca 4500

tctgggtggg actctgtttt atcatcttgg tgtcacggct cctggcccac ggcctggcac 4560

tgtctgaatg tttgggcgat gggtgaaggt gagttgagag atgggtggga attggggaca 4620

ataaaccgtc ccacctgctg gtaccttgac cctgggaaag gctgggaagg tgagatcctg 4680

gggcagatgc ccagcctcat gggtcatga atgggcaagt ctgtgaatac tcagaggccg 4740

cccccagggc agggcttctc aaacggcccc tctgaagccc cttcccccat ctctccaccc 4800

tttgaaccca gaggccatgg gcaccggaga cctggacctc ctatcagcct tccgccgcac 4860

gtgagtggcc atcctcaggg cctgggggag ccagtgctgg agtctgaggc ccttcaggt 4920

ctcgactggg ggtcagggct ggggatgatt tgcggggcgg aactgaagtc agtgggggct 4980

gcaggggaga gcaagcttcc caggcaggtg aggatactga gtctaaccccc cacaggagcc 5040

tggctggagg tggtcggcgg atcaggtacc cctgccccag gccctacctg caccatccgc 5100

agacccccag gggtctgaga cgggtgggag cccaatctgg ctagtgtccc tttctccctc 5160

cccactccac tcccatcctg ctcctaatcc ctttccagtc cctcactgca gcctcactgg 5220

gggtccctct ccatagtgat agccatgagg acactgggat tctggacttc agctcactgc 5280

tgaaaaagag gtgagtcctg ggtggcagtt cctggggcat ctggtgaggc ctggctccaa 5340

atcccagcac tgtggtttgc cagctgtgtg accctgagca caacactgca cctctctgag 5400

ttggtttcct tgtctgtcga ggggatgatc ccagtgtgga aggagttaaa gggctctgca 5460

gacattatgt cccgtcaaca gtcatcctca cagtgtcccc cacggccact ccacacttga 5520

ggctgcaaaa gctccctccg taaactccgg ggacccagaa ggaacagaga gagggtcccc 5580

agagctgcag ggtctaccag gtcggcccaa ctgacttatg ttctctctgc cctctctctc 5640

cctctccccc ggcccctcca ttatggctgc tgctgctgtg gcccagagag taagaatcgg 5700

ggtctgggtg tttgggggcg gcagggcgct ggtgggtgca gaggggattc ccgtcccctc 5760

tctgagaggc tgtggtgaga gactcagagc agggtgcggc tccccacgga caggggtgag 5820

tctctgtgcc ttgggcttct cctgccacccc acctatctgt cagagcttag gagggataag 5880

aaaaagtaat gcacacagga ggtcctgcca gagcctccgg agcagcatgg gtgccccaca 5940

acacaggggag tgcaagtgcg gaaaataggg aggaagtaaa ggcccatggg tggggtccag 6000

gtctttgagg gaaggtggcc atacctctca tgtgccacct accctttctc ccaccccccac 6060

ccctggagca gcagtttccg gaccccgagg tgagtgccca acatagcaca gcaccctcat 6120

cacccctaat tctgccagga gcccctccag atccccagcc cctcttcagc tcccttggcc 6180

cccagtcctg cctggcctgg gacccagagc agctccagct gccccaccag aagggaaggg 6240

gcagagggac aggggtggct acagctcctt ggtcctgggc ccaggaagcc ccgcccaggg 6300

ggctgcagtc ttgcccccgg ccacagccta gactgcggga cacagggact cgaagctgga 6360

ggcaccagca gaggaggacg tgtgggagat cctacggcag gcacccccat ctgagtacga 6420

gcgcatcgcc ttccagtacg gcgtcactga cctgcgcggc atgctaaaga ggctcaaggg 6480

catgaggcgc gatgagaaga agagcacagg ttagcccttc ctcagaggga gaggaagagg 6540

gcacaggcta gcccttccct acacaggaga ggagaggggca taggttagcc cttccccaca 6600

ggggagagga gagggcacag gttagccccc cacctcgtcc acatacatgg aaagagcgga 6660

gtccggcttt agcacagaga ctcacaggaa acaagagccc aggttaaccc ctccttgtgt 6720

agaaaaagcc aagaatgccc aggttagccc cttaccacag agatagaaga gaagagcaaa 6780

agttaggcca ggacagccac aaggaaaagg acagagctca ggttaaccct cctcaccaag 6840

gaaaaatcag gctacacttt ctttctctac cctcttctga aaagaaatga agcccggtta 6900

accctctcca cacccaaaag aaaagggaag aggcccgcta agcctggaga gccacacaca 6960

ggcaaagaac agagcctggc taatctttct ctcaggcaaa caaagcagga tggcttgggt 7020

taactaccct cacccccagg cagatggaaa gaggagagcc caggaatagc ccttctccac 7080

tgaggggttg acagggcggg tgtgaccggg aaagcagggg tctcacatgc cttctcccca 7140

ccatgtctgg gtctagcaca gggcctggca cacacctcga gcgctctata aacaccgact 7200

gaatggaggg gtcagcttcc atgcaggata tgggagacaa gcccctccca aagcacacac 7260

acaaaacaca cacgatcatg gggacggggga gaggagctgg ttagtccctc cctgctgacc 7320

tcccccaggc cccttatccc tctgccctgc ctccctgttg tctcagcctc actgggtttc 7380

ccatcacccc atcgctaggt ggcctggccc ttaccggcac gtggcccatt taagccaggg 7440

atcccctgaa tgccacaact gtctctggta ctcctgaact gctgagccca gaagggcaca 7500

agccannnnn nnnnnntggg caacagaggt gctagtggag cgctgggaag gagtacacac 7560

aactcagggt gctgagagaa gagaaggcac agcgtcccag ccccagttct ctgacaagtt 7620

ctttctttct ggaagcccta acccatcctg agctgagcag gtggagagga ccccaaggaa 7680

gacagaagga caagtcagac aagacagac aaggaggtgg gttgacacat aagacagat 7740

aaagaaatga agacaccatg cctgtaatcc cagcactttg ggaggctgag gcgggcgcat 7800

cacctgaggt caggatttca agaccagccc ggccaacatg gtaaaaccccc gtctctacta 7860

aaaatacaaa aattagctgg gcgtggtggc acatgcctgt aatcccagct atcgggaggc 7920

cgaggcagga gaatcacttg aacctgggag gcagaggttg cagtgagcca agatggcggc 7980

cattgcatcc agcctggccc acagcaag atttgtccga aaacaaccaa cagacacacc 8040

aaccaaccta aagaaataga gacacagaga agtggaagcg gggcggcaca gaggggattg 8100

gaggggggga cctgggcccc agccacagcc acagtagctt ggcctggggg agcagggtgc 8160

gggcggtggg gtggccgggg ctgaggggtg gtgctcagcc tttcagaaga agctggagcc 8220

ggcctaccag gtgagcaaag gccacaagat ccggctgacc gtggaactgg ctgaccatga 8280

cgctgaggtc aaatggctca agaatggcca ggagatccag atgagcggca ggtgcagcct 8340

ggggtgggga gggggctcgg gtggaggtgg ggacccccat agccttgcct cctgcccctc 8400

tccacatgcg tatctctgac tcggtgtggc tctcagcccc atctctctgg gcctaatttc 8460

ccatcctttt gctcctgccg gtccctctct ctctctcctt ttgtctcggg gctcacttcc 8520

tcccgtcggg aacacttcaa cggccccttc tgttctacag caagtaagtt cccctctgga 8580

tggcttgggg tggggggcac agggattatc acggggcgca ctgggcccgg agggggtgtcc 8640

gcagctttcc tgccacttcc ctgcggcccc cacccaggta catctttgag tccatcggtg 8700

ccaagcgtac cctgaccatc agccagtgct cattggcgga cgacgcagcc taccagtgcg 8760

tggtgggtgg cgagaagtgt agcacggagc tctttgtgaa aggtggggcct gggacctgag 8820

gatgtgggaa cctggggagg agatggcctc aggggagcca accctcatgc tcaccctgcc 8880

tggacagagc cccctgtgct catcacgcgc cccttggagg accagctggt gatggtgggg 8940

cagcgggtgg agtttgagtg tgaagtatcg gaggaggggg cgcaagtcaa atggtgagtt 9000

ccagaagcac ggggcatggg tgttgggggc atctgcccag aagaggccac agcacttgcc 9060

accccaccac ccggctaggc tgaaggacgg ggtggagctg accccgggagg agaccttcaa 9120

ataccggttc aagaaggacg ggcagagaca ccacctgatc atcaacgagg ccatgctgga 9180

ggacgcgggg cactatgcac tgtgcactag cgggggccag gcgctggctg agctcattgt 9240

gcagggtgag cctggctggg ggggcacatg aggctttagg gcttggaccc ctcagccccc 9300

accccaccta gccctgtagg ggagcaggca aacctgggct caaggcctct gtgaccttgg 9360

gcctttgaca gcctaaacct catccttctc atctctaaaa tgacgatgct gaggtgaccg 9420

ctccagaggg tactgagagg gtcatacggg cctggtgagc atcaccaccc ctcagacact 9480

tgaggttcct tatgtgcact gcaccatcac aggcaaacat ggcacacaca ggcacacgtg 9540

ttttcacatg cccacatgca cccagacacg tgcgcaccag cgcccatggg cccacacgcc 9600

ctccacagggg attcacgcca cacccacaca ccctcccgag ctcaatggct ctgccctgcc 9660

ctgcagaaaa gaagctggag gtgtaccaga gcatcgcaga cctgatggtg ggcgcaaagg 9720

accaggcggt gttcaaatgt gaggtctcag atgagaatgt tcggggtgtg tggctgaaga 9780

atgggaagga gctggtgccc gacagccgca taaaggtgtc ccacatcggg cggtgagtgt 9840

gcagggcagg tggatgggac aggtggagac tgagatggag acagagagag acaggggag 9900

aaacagagag agagagataa agacaggtgg acagagaggc agatgcacag agacaggaca 9960

gagactgaga cagagaccca cagagaggga gaaacagaca gacccaaaaa gacaaagaga 10020

cagaaagaca gaaatagttc tatagagaca gaaagatatg cacagggagg cacacacata 10080

ggagagttcg tcagagacag agaagaacag agggaacagag acaaagaggc agcaggggag 10140

agacacggac ggggtcgaga gctccccagc accctccccc agcttgtgac cagggcaggg 10200

caggaggcca ccaaggaggc cggctctggc cagaggccct cactgcctct cccgccctct 10260

tgctcatccc tggagcaact gggctggcct gctgggccca agttcacaga gattaatgag 10320

acccagcccc tgcccagtct ccaagggccc agccaaggag atgctttgat gtagaaatca 10380

cttttacgct gcctacctct gaaggcacat taaaaagctg gaaaattaga taatatagaa 10440

aaaaaaaggc atttttaaaa atcagaatac caacaagcca ggacaaggtg agggcctcct 10500

gggaccctgg ctggggtatc tggcaaggcc agggggtgtgt ggcccagtgg ggtcccctga 10560

gccactgctc ccctgcaggg tccacaaact gaccattgac gacgtcacac ctgccgacga 10620

ggctgactac agctttgtgc ccgagggctt cgcctgcaac ctgtcagcca agctccactt 10680

catgggtgag cctgctccag ggtagggtgg gtcggggcag tggggccagg agcccctgtc 10740

actgggccct gcctttgccc ccgtgctact tgctcttcct tctcttgcag aggtcaagat 10800

tgacttcgta cccaggcagg gtaagtcttg gggcccctga gtcttggttc tgctcacttt 10860

cccgcaaccc accacgcag gcctcccctc cactcacaga gggaaaagca aggatcaaaa 10920

ttgggggtcc tggggcccag ctccagcttt gccactcatg accatgggtc aattataatc 10980

tctgtgagcc tcagtttcct cttctctaaa atgggaaccc tgatagtgcc acctcatagg 11040

atagttataa agatgaggtg acatgatgta tcagacctgt caagtgctgt cacactgtga 11100

ctgttattat tacatctttt cttcttcttc cttttttttt atttttattt tttttgagac 11160

ggattctccc tctgtcgccc acgctggaga gaagtggcgc gatctcggct cactgcaagc 11220

tccgcctccc aggttcacac cactctcctg cctcagcctc ctgagtagct gggactacag 11280

gcgcccgcca ccatgcccag ctaattattt tttgtatttt tagtagagac ggggtttcac 11340

catgttagcc aggatggttt cgatctcctg atctcatgat ctgcctgcct cggcctccca 11400

aagtgctggg attacaggcg tgagccactg cgtccagact gctttgcttt gtttaactga 11460

gggcagattc ttgatttatt tggccaggga acccttttaa tgtgtgcatg tgtgcgtgca 11520

tgtgtgcgcg cgtgcgtgtg tgtgcatgtg tctgtgtgtg tatgtatata tgtgtgtctg 11580

tgtctgtgtg tgtgcgtgta tgtatgtgta tttgtgtctg tgtgcatgtg tgtgtctgtg 11640

tgtgcatgtg tgtatgtgta tgtgtgtctg tgtgtatgtg tatatgcgtg tgtgtgtgtg 11700

tattgtgtgt gtgtgtatgt gtgtggcaag acactgatgt tcccccagtt ttcaaaatgc 11760

tatgctaggt gacctgacct gaatattaca agcctcccag ccaacgtcct ggggtgatgg 11820

gaatgttcca gatcttggtt ggattaataa ttgcatgggt gtattcatct gtcagaactc 11880

agcctactgt cagatctgag cataaactat acctcaatac aaatctaaaa aaccaataac 11940

caggggccca ggctcttgag cagccccgcc ccagtgacct gtgctcctcc tggctctccc 12000

gtttctctga actacattgt gtcttctgca gaacctccca agatccacct ggactgccca 12060

ggccgcatac cagacaccat tgtggttgta gctggaaata agctacgtct ggacgtccct 12120

atctctgggg accctgctcc cactgtgatc tggcagaagg ctatcacgca ggtactgtgg 12180

gtccctcctc agtctcccca tctataagat gggtgtgtgg aaccagccaa gtgctaggac 12240

ctgctgcaca ccccaccac atgtgcacgc cctgctgttg gctaccccat gggagagact 12300

ggtcaggggg caggcaaggt gggcagtgtg ggtcaggggg tggaagaatc agcagagggc 12360

cgcttccagg ctgagtcagc tcctctgctc cctacttccc tcctgccctg ttcccagggg 12420

aataaggccc cagccaggcc agccccagat gccccagagg acacaggtga cagcgatgag 12480

tgggtgtttg acaagaaggt gagtgagact gaggtcagga ggaggatcgt ttgtctttca 12540

tcactttcta cctacgtggt gccggccctt ggagaagtct ccataggtgc cacacttggt 12600

cgggtgtgtt gctgccccca gcagggctcc ctgtgggacc ctcaacacctc agccagaggg 12660

tctctgttga agttgctgat gcccatcaca aggatgtgga caccctctca ttaccaggaa 12720

cccctcatga ctgggctgct ccctggctcc cttcatccta agctgaccat gaccttgacc 12780

ttgtgctccc tctcaactca ggagtagttt ctgagacact tttctgccag tgtagctgcc 12840

ggggtcttgc tcctgcctgg cactgttggt tcttctgaag gctggggtct tgaggggtcc 12900

tcatgatagc tcacctgaga ggaacaggtg gttcccccag gtcccgtgcc aaatcaagaa 12960

aggcgagccc ctgccttcag tcggtgccca gagatgaatt gtactagaat attgtggatt 13020

cagtcttccc acagactgtg ttccccccat ggacccccgg gacatctggc tgttgggagg 13080

ctgcaaggac ctcctgggtc tgacttggat ctcaccccaa ctctgcaccc ccccagctgc 13140

tgtgtgagac cgagggccgg gtccgcgtgg agaccaccaa ggaccgcagc atcttcacgg 13200

tcgagggggc agagaaggaa gatgagggcg tctacacggt cacagtgaag aaccctgtgg 13260

gcgaggacca ggtcaacctc acagtcaagg tcatcggtga ggccggccgg ggtccaagct 13320

ggagaacaca gaggggcagc cccaaggagg gcccatccgt tcgctcattc agccactcga 13380

caaacatcac gggcatgctc cctgagccga gcggcaacag gacaggtttt tctgcccaaa 13440

aggagccccg ctctggggga gatggaggca gatgcatcca gcagatttcc tgcgacactg 13500

ctgtacatga gccatgtcca gagtgccgtg ggagcactga gcagagagcc atccctgtgc 13560

ctgggggtga tgcctttagg aggctgagaa gatggtgagg aaaggcatcc caggcagagg 13620

aaactgtgta caaaggggtg gaggagggaa agggcctttt gggtgtggag aatgaatgaa 13680

acatgtcagg agaggactgg gtggggttgg gaggatcaga gccaggacag gggtgaatat 13740

gggcagtgag aagccttaaa gggttgacgg agggctggcg acatgatcag aaatgcatcc 13800

tgttggagac cagcctggac aacacagtgg agactccata tcactaaaaa aaaaaaaaaa 13860

aaggaaaaga agaaagaaat gcattctaga aagatctctt tggatctggg ggtggtgggg 13920

gcagcctgtg gcggttagtt ggagtgggaa ggggacgagc aacgttactc aaggccctga 13980

gcggggcagg gctgatgtgg gtccatccca ccccatccag acgtgccaga cgcacctgcg 14040

gcccccaaga tcagcaacgt gggagaggac tcctgcacag tacagtggga gccgcctgcc 14100

tacgatggcg ggcagcccat cctgggtgag tgcaagggca ccggatggag gtgtgagggc 14160

gccaaacaga tccgagggaa ggtggtgtgg ggatgcctgg gttccagacc agagctgcca 14220

cctcccctga gccaggctac atcctggagc gcaagaagaa gaagagctac cggtggatgc 14280

agctgaactt cgacctgatt caggagctga gtcatgaagc gcggcgcatg atcgagggcg 14340

tggtgtacga gatgcgcgtc tacgcggtca acgccatcgg catgtccagg cccagccctg 14400

cctcccagcc cttcatgcct atcggtgagc ctgcctggcc tggtcctgcc ccccgccccc 14460

tccccagtta aaaacctcca ataatagcag gtgctctgca ggcagccatg ctagacactc 14520

acatccacag tctcctttca tcctcgactg gggatactca gccgcatttt acagatgagg 14580

aaacaggctc caagcagtta aatgacctgc ttgagatctc ccagctggtt gggtggagc 14640

tgactctcac actacgccga cccctagttc acgtttcaag cctctctttc catgtctaca 14700

aaatgggaat aatgacactg cctatctcag agagctgtta ggatgatatg ggatgaccag 14760

gcaggccatc tggcatctaa caggtgctca ataaatgttc attgtcccct gttctccaca 14820

cccttcaacc cagggttttc acctatggga tggggcaagg gctgtagtta gtggaggggt 14880

ctcaggggca tctgtctgtg gaaagagtcc tggaccaaga ctcagagatg gcagaggcag 14940

ctccatggtc cccggggggc caactggagt catccctccc tgaagcctca gcttcctcat 15000

ctgtataatg gggtctgagt gggcagaaca gtgttgagac tgtgcaggct cagagacacc 15060

tgtcccacta ggtggtgatc ggggcagctg ctgtgcaggc agcagacctg gccagatggg 15120

cacacttgga gatgcagatc ggctgtggga gaggtcagct ctcttttagt tttttttttt 15180

ttttttttgg agatggagtc tcgctctgtc acccagactg gagtgcagtc ctggcgcgat 15240

ctggggtcac tgtaaccttc acctcccagg ttcaagcaat tctcccacct cagccttctg 15300

agtagctggg attacaggca tccaccacca tgcccagcta atttttgtat ttttgtagag 15360

acggagtttc accatgttgg ccaggctggt cttgaactcc tgacctcagg tgatcctccc 15420

acctcggcct cccaaagtgc tgggattaca ggcgtgagcc accgtgcctg gctgctgttg 15480

ctgtttgttt ttaagccaat ttacagacat gtcttgtaac agggacaaaa tattccttaa 15540

agtggccaaa agccagctga acaggaagtg ccccctatgt gaccagtggg cagttcagag 15600

tctagggcat ggatctccag cttccccagg cttgctcaga cccctctctg acctctcctc 15660

tgcccaggtc cccccagcga accacccac ctggcagtag aggacgtctc tgacaccacg 15720

gtctccctca agtggcggcc cccagagcgc gtgggagcag gaggcctgga tggctacagc 15780

gtggagtact gcccagaggg ctgtgagtgt ccccgccccc aacccccctc cccaaaggaa 15840

gaacatgctc accttgccat tgagcagatt cacctgtagt caagtttttt aggcccactt 15900

ttgccgaaag ttgaggacac ccagagaaat atctgtcctg ttttcagaag taaaaaagct 15960

tgcctagtga aaaacaaatg cagagtaaag ctgactgtaa cacttctttg agcagtgcga 16020

aatcagcaac tacattttaa aaacatattt aaggccaggc gcgatggctt acgcctgtaa 16080

tcccagcact ttgggaggcc aaggtgggcg gatcacctga ggtcgggagt tcgagaccag 16140

tctgaccaac gtggagaaac cccgtctcta ctaaaaatac aaaattagcc gggtgtggtg 16200

gtgggcacct gtgatcccag ctacgcggga ggctgaggca ggagaatcac ttgaactcgg 16260

gaggcagaag ttgcggtgag ccaagatcat gccattgcac tccagcctgg gagacaagag 16320

cgaaactcca tctcaaaaat aaataaacaa ataaaaattt aaaaaaacat atttaaatcc 16380

tggagattcc tatcagagga gtgggcagtg ggagtggggt gtcagtggtg acacagcctg 16440

tggccttgcc tccccctccc cacccccagg ctcagagtgg gtggctgccc tgcaggggct 16500

gacagagcac acatcgatac tggtgaagga cctgcccacg ggggcccggc tgcttttccg 16560

agtgcgggca cacaatatgg cagggcctgg agcccctgtt accaccacgg agccggtgac 16620

agtgcaggag atcctgcgtg agtgcccctt tgtgcagtca caagacccgc cattgacacc 16680

ccaggccctt ggtgtccagt ggaggtgggg acagttgggg aggcccagaa tgctctgccc 16740

agaacgctgg gcagagacag gaggggttgg agccatcgtc catgcgctag gcctagggaa 16800

aaagtgcaga gagggtggcc aggcacagtg gctcaagcct gtaatcccag cactttagga 16860

ggccaaggca ggcagatcac ttgaggtcag gagtttgaga ccagcctggc caacatagtg 16920

aaaccctgtc tctactaaaa atacaaaaac tagccaggtg tggtggcgcg tgcctgtatt 16980

ccctcaacta cttggggagc tgaggcagaa gaatcgcttg gacccgggag gtggcagttg 17040

caatgagctg agatcacact attgcactcc agtctgggtg acagagtgag acaacgtctc 17100

aaaaaaaaaa aagtgcagaa agggctctct tggtgtatgt cagtccctta gttctggaac 17160

agggaaagta aaagccacag taccaagct ctcttccagg agtgtctcag ctccaacagg 17220

taaatattgt ctatgtagtc agattggtcc ctctgggatc cacttcccat tcctttgttc 17280

cttgtcccaa caaagggtcc atcttttgta aggattggtt ctcaaagctg gaacgaggca 17340

gcaaactcag gtggtaacaa cacagggcat agggtgtcaa agcccaggct tacaccctgg 17400

tctatgactt actagctgtg tgacttcagt caagttgctt accctctctg atccctgcct 17460

tttgaagtgg tgaggctcgg ggaagccagc tgggcttggg cagcctggag ttgctgtgtt 17520

agcaggagct aaaggaggcg cagggcccag caggcagctt ttttcttagc tgcagctctc 17580

tgggccttgt ctcaagggag gttggagctg tggagcccct cagagctgtg gcgggccctc 17640

acttagctac ccactttata cccacagaac ggccacggct tcagctgccc aggcacctgc 17700

gccagaccat tcagaagaag gtcggggagc ctgtgaacct tctcatccct ttccaggtgg 17760

gactggcccc cttccctgtc ccccaggggga gagaggctat agtgtgttgt tcccatccag 17820

tggactgatg tctcagggct ggcctgatct gaggtaccag aggctggggt ggcggccggc 17880

ccttggagtg atccaggtc agggttaagc ttttcctccc ctcagggcaa gccccggcct 17940

caggtgacct ggaccaaaga ggggcagccc ctggcaggcg aggaggtgag catccgcaac 18000

agccccacag acaccatcct gttcatccgg gccgctcgcc gcgtgcattc aggcacttac 18060

caggtgacgg tgcgcattga gaacatggag gacaaggcca cgctggtgct gcaggttgtt 18120

ggtgcgtggc caaggcctcc ttgagccccc ttgtttccct tccctgggct gggctgctgg 18180

ggccaagggt ggggccacgg ggacccaacc cacagctcac attttccagt ccactgcccc 18240

agcaggcctg gccctggttg gcaggggtgg ggtggtccct ggagccagtg accccctgct 18300

cactgtcagg aggcgtggtg acccaactgg gtctgtctcc cgcagacaag ccaagtcctc 18360

cccaggatct ccgggtgact gacgcctggg gtcttaatgt ggctctggag tggaagccac 18420

cccaggatgt cggcaacacg gaactctggg ggtacacagt gcagaaagcc gacaagaaga 18480

ccatggtgag cccagggtct gggtcccca cgtgcaccct gcctaggccc ggcagaccca 18540

ggccgcagct acccttcact gtcctcaccg tggacccctc accctccctc tgctgatctg 18600

aatccctcca tagtaggggt ggagatgcca agggcaccca ggagaggctc tcggcatcag 18660

gaagggcctg gcccagagat gcctcccctc cctccctgcc cccaggagtg gttcaccgtc 18720

ttggagcatt accgccgcac ccactgcgtg gtgccagagc tcatcattgg caatggctac 18780

tacttccgcg tcttcagcca gaatatggtt ggctttagtg acagcggc caccaccaag 18840

gagcccgtct ttatccccag accaggtgct gtaccctcat tcttccaacc aggggctggg 18900

gcagggaggc tgtgggaaca gggagagggg cctagctttg tgtggccctc tcggtaccaa 18960

gtcctgtcac caacaggcat cacctatgag ccacccaact ataaggccct ggacttctcc 19020

gaggccccaa gcttcaccca gcccctggtg aaccgctcgg tcatcgcggg ctacactgct 19080

atgctctgct gtgctgtccg gggtagcccc aaggtaggga actttaggcg ctggtccagg 19140

ccgaccagga ggcagggctc acaggccccg acgttgagca gtcctctccc cagcctgtct 19200

cccctgcttt ctctccacct tggggtgtat ggaccgggtt cctccccctg ggcagggcca 19260

tggtactcac tcttggttcc atgtttgttt ccagccttgg gcatagtcag ggactctcgt 19320

gggacccccc gagtagaaac acagatgtgt ctccctgggt ccctgccagg tcccctctca 19380

gcctggatgg cttccctccc tctctttacc ttattatag cccaagattt cctggttcaa 19440

gaatggcctg gacctgggag aagacgcccg cttccgcatg ttcagcaagc agggagtgtt 19500

gactctggag attagaaagc cctgccccctt tgacgggggc atctatgtct gcagggccac 19560

caacttacag ggcgaggcac ggtgtgagtg ccgcctggag gtgcgaggtg aggacccctcg 19620

gggccagggc ctgggatg ggaagagcgg gcggcagtgg ggaccctggg ctttgctccg 19680

ttgtcctcgg ccaagcacct gccctggctg caagggcacg gaccctgcca cgcccagatg 19740

ggcaatagct tccagaaggc tgggaggaca cagtgacatg gcctcctctt ctgcagtgcc 19800

tcagtgacca ggctggctcc tggggatggc caggtatgtc cacccgacac ccagggcaca 19860

agtcagtggc tgagccctgg cccttggtcc tggggcagcc atcagtaaat gggaggctgt 19920

aggggccctc cattcactcg taagataacc tgtgttgcag gtacaaccgg atgccagccc 19980

cgtgccagga gcctggaggg aagttgggga aacccctccc tactgttgga tgtatgtgtg 20040

acaagtgtgt ctcctgtgct gcgatggggg atcagcaggg cagttgtcgg gcagtcctga 20100

gtgggtgttg cacagactgg tccacagggc tcctgaagga agcccctgga tctttggggt 20160

aaaaggaggg tggcctcaag aaacaatgtc tggggacagg cctttctggc ctgctatgtc 20220

ttcccaatgt ttattgggca ataaaagata agtgcagtca cagagaactc actcttcatc 20280

acctgagctc accatagtct aatagaaaac aaccgaaacc aaataattca agcactgctt 20340

attacaattt tactgggtct ctattttacc ctcctacaag cctcagagta cttcgagtct 20400

cccttcacca tttccgacgg catctacggc tcaacatttt ttgtagccac aggcttccac 20460

ggacttcacg tcattattgg ctcaactttc ctcactatct gcttcatccg ccaactaata 20520

tttcacttta catccaaaca tcactttggc ttcgaagccg ccgcctgata ctggcatttt 20580

gtagatgtgg tttgactatt tctgtatgtc tccatctatt gatgagggtc ttaaaaaaaa 20640

aaaaaaaaaa aaggaattct cg 20662

<210>3

<211>6592

<212>DNA

<213> cTnT gene

<400>3

gaattccaaa ttccacttct gaggctctaa tgccagccta ggatctgaca ctgagtctca 60

gggccttgac tttggccccc tgttcccagg caccctgtgg ctgactaggg ctggggtgtc 120

tcctcaggag ggcctcagca cccagaggat ggttcaagta agcaagttgt gtccaccctt 180

ctgtgtgtcc aagcgtcttt cctgtcctat gtccgtctcc agggctggga gagaaaggcc 240

caggagtgct ggacagagga ggccagaaac cagattcctg ggtttccaag tggacgaggg 300

atggatttcc cagtgctatt gactgagagc tagccccgag cacagctgca tcctcaaatt 360

aatcaacaac atttattgag tggcctggac tcctgggtct gaggggagag ggattggggt 420

cctggactcc tcggtctgag gggagaggga ttggggtcct ggactcctgg tctgaggggag 480

gaggagctga gggtgcagac tcttgagtct gagagaggaa gtgtgctggg ggcctgcctg 540

gactcccttg tctgaggggag gggctggggg cctggaccccc tgggtctgag ggaggagggg 600

ctgggggcct ggactcctgg gtctgaggga ggaggggctg gggcctggac tcttgggtct 660

gagggaggag gggctggggc ctggactctt gggtctgagg gaagaggagc tgggggcctg 720

gactcttggg tctgagggag gaggggctgg gggcctggac tcttgggtct gagggaagag 780

gagctggggg cccagacact tgagtctgag ggaaaagggg gctgggggcc tggactcctt 840

gtgtgaggga gtggccaggg tctcccaggg tcctgggact gagggaggag gaggttggga 900

ggtcgggcat tctctgggtc tgagggagga ggggtctagg actccaggat cagagggagg 960

gtccggattc ctgtgcccgg aggtggctgg tgatccagct ggagcagggc ccaccccgcc 1020

cctcacgcgc ccccgttatc tggcatagtg ggcaggggga ggaggaggtg acagtatatt 1080

tagtctgtgt cctcgccctt tatctcagtg tcctcgggga gtctcaagca gcccggagga 1140

gactgacggt ccctgggacc ctgaaggtca cccgggcggc cccctcactg accctccaaa 1200

cgcccctgtc ctcgccctgc ctcctgccat tcccggcctg agtctcagca tggcggatgg 1260

gtgagtgatg ccccaaggca gtggggagtt gggggcgacc tcccgggttc ccaagagggg 1320

tcgcagctga gaggctggac ccaaggcctg cgaggatggc gtagggactc ttccctgaag 1380

agaggaagtg ggtttgcgag tcagactcct ggaacccaag gaaggggaag ctcggtcccc 1440

cgacctcttg ttcatagggg actccagggg tcccttagga gacaggacac agcccaccac 1500

taacccccct ccttggtttc tctccttcca ggagcagcga tgcggtgaga gcagcgggct 1560

aaggcgtggc tgggaccccc aggtccaggg tgggccctgc agtgaggggt ctgggtcggg 1620

aggctgcagc ctagcagagg gtgcggtacg gtaagggctg ggtggggtct tggtggtcat 1680

gggtcccca ctcctcctaa cccaggctag ggaacctcgc cctgcaccag ccccaatcag 1740

acgccgctcc tccaactacc gcgcttatgc cacggagccg cacgccaagg tgggacgggg 1800

cttcctgggg gcagagtaca ggcgccggag ggatccaaga ccctgggagt ggggggagga 1860

gccagggctg cgaagggggc ggggactacg cggaggggct tcaggggcgg agttttgcag 1920

agggtcatgc tcggattggt gacagcagcc tgcgggcgga actccgttgc cctcggactt 1980

gcttagggat agatgggaag tgcctatcca aaggaagaga cccagattgg tggatgggaa 2040

tgaggggcgg ggcctcccgt agactcaggg ctcaagttgg acgtgggccc aaatctggac 2100

cggctgggtt tgctgggggt gtcttgaggt cccctccacc gtcgtctccg aatccccctc 2160

catgatcctt ccttgctcca tctcaccctg gcagaaaaaa tctaagatct ccgcctcgag 2220

aaaattgcag ctgaaggtga ggacgggcgg gactgggaaa gagcaggcag gtgctcaggg 2280

ggcggacgtt gagaatgggt ggggctttcg gggtaggtgg gcggaagtgg gcggggtttg 2340

gccgctcggg gcgtggcttt agcggggagt actgctcggg gtgggacggg gccttggaac 2400

agtggagacc aaactggagg gtttagaagg gcagaggcgg ttccccacgc ctggtcttta 2460

tcctgaagcc ccgggtggcg tgcgcttccc tcccaccccct ctgcagactc tgctgctgca 2520

gattgcaaag caagagctgg agcgagaggc ggaggagcgg cgcggagaga aggggcgcgc 2580

tctgagcacc cgctgccagc cgctggagtt ggccgggctg ggcttcgcgg agctgcaggt 2640

accggctccc aaggatgcga ggtttctagt cccggaattc agcagtacag cctctatccc 2700

ctcttctgct cgggacccag gcgtccaata tggctgtcct tacccaatta tatatggttc 2760

gtgggactcc tggcccctaa caccctttgt gtgcaggtct gtggagtctt ggctccaacc 2820

tactccttca ggaccatgtg gccctcctat cccagacaga agcccaagcc ccagcccctc 2880

ctccctcaga ccctggagtc caggccccag cccctcctcc ctcagaccca ggagtccagt 2940

ccccagcccc tcctccctga gaccctgaag tccaggcccc agcccctcct ccctcagacc 3000

ctggagtcca ggccccagcc cctcctccct cagaccctgg agtccaggcc ccagcccctc 3060

ctccctcaga ccctggagtc caggccccca gcccctcctc cctcagaccc tggagtccag 3120

gccccagccc ctcctccctc agacccagga gtccagtccc cagcccctcc tccctcagac 3180

cctggaatcg aggccctcag cccctcctcc ctcagaccct gaagtccagg ccccagcccc 3240

tcctccctca gaccctgaag tccaggcccc agcccctcct ccctcagacc ctgaagtcca 3300

ggccccagcc cctcctccct cagacccagg agtccagtcc ccagcccctc ctccctcaga 3360

ccctggagtc caggccccag cccctcctcc ctcagaccct ggagtcgagg ccccagcctc 3420

tcctccctga gaccctggag tccaggcccc agcctctcct ccctcagacc ctggagtcca 3480

ggccccagcc cctcctccct cagaccctgg agtccaggcc ccagctcctc ctccctcaga 3540

ccctggagtc caggccccag cccctcctcc ctcagaccct ggagtccagg ccccagcccc 3600

tcctccctca gacccaggag tccagtcccc agcccctcct ccctcagacc ctgaagtcca 3660

ggccccagcc cctcctccct cagacccagg agtccagtcc ccagcccctc ctccctcaga 3720

cccaggagtc cagtccccag cccctcctcc ctcagaccct ggaatcgagg ccctcagccc 3780

ctcctccctc agacccagga gtccaggccc ccagcccctt ctcctccctc aaacccagga 3840

gtccaggccc ccaactcctc cctcagaccc aggagtccag gcccccagcc ccttctcctc 3900

cctcagaccc aggagtccag gccccgagcc ccttctccct cagacccagg agtccaggtc 3960

tccctgtttt tggttccccc aacaacacac accacgttcc tcctccagga cttgtgccga 4020

cagctccacg cccgtgtgga caaggtggat gaagagagat acgacataga ggcaaaagtc 4080

accaagaaca tcacggaggt gggacgcatg ggcagctcgg gtaccttcgg ggtagggtga 4140

gatggctggg acttggtctc tgcctgaccc cttgcagctg cttttggctg cacatcccag 4200

gagacccagg acaactgtga gcctggcagg gctggggcag aaggatgagt acaatatagt 4260

caaggaaagc tgttctaggc agagggaaca gcacatgcaa ggccatgggt tgggaaacag 4320

aaaataagtt agtgaacatg ctcagggcat cacatgttgg taaattagct caggcactgg 4380

ccagggaatt gtgatttgca tgtagctgga ccaggttatg ccagtggttt tgagaggtga 4440

ggctggagca tatgaggagg gggattcagt tccaggatta gaagcctaga ctgggagcct 4500

aagccgggaa gagactggta aggcctcggt actggaagac gagataagga gaataaaaaa 4560

ggagtgtagg atggaggagt tgggtgtgcg ggaaatggaa ggagaagtac ccaccccctc 4620

gtgtgccccc agattgcaga tctgactcag aagatctttg accttcgagg caagtttaag 4680

cggcccaccc tgcggagagt gaggatctct gcagatgcca tgatgcaggc gctgctgggg 4740

gcccgggcta aggagtccct ggacctgcgg gccccacctca agcaggtgaa gaaggaggac 4800

accgagaagg tgagtgtggg ctaaggccag gaaagaggat gctgaggggga agggctgtgg 4860

gtgccaacaa ccctaggcct gagggcagat ggtgcttgga gttggaggta gaagcagcta 4920

gtaaggggtc ctcagaaatg caagaggaag acaggaagta gaaggggaag acaggaagtg 4980

cattagggct acaggaagtc catgtaagag caaagaggta catgaaggtc caggtgcagt 5040

ggcttatgcc tgtaatcttt gaggtttaaa aaaaattttt tttaaatttc cttttttttt 5100

tttttgagat ggagtctctg ctgtccaggt gcagttgctt atgcctgtaa tctttgaggt 5160

ttaaaaaaaa ttttttttta atttcctttt tttttttttt ttgcgatgga gtctctgctg 5220

cccaggctgg agtgcaatga caccatctcg gttgctggag tgcagtgaca ccatctcggc 5280

tcactgcaac ctctgcctcc caggatcaag tgattctcat gcctcagcct cccgagtagc 5340

tgggaactaca ggtgcacacc accatgttgt tgttttaatt tctacaaatt tcttttttaa 5400

attagccagt cacggtggtg gcatgcagct tctcaggagg ctgaggcagg aaaatcgctt 5460

gagcccatga gatcgagggt gcagtgagct atgactgcac cactgcactc cagcccaggc 5520

aacactggga gactccatct caaaacaaaa caaaacagaa caaaagttag ctgggcatgg 5580

tggcacacac ctgtggtccc agctcctcag gagtctgagg tgagaggatg gcttgagccc 5640

aggaagttga ggctgcagtg agccgagatt gcacgactgc actccagctt ggatgaggca 5700

gccagaccct gtctcaaata ataataataa taaaatagaa caattataac agattgtaat 5760

aaaactcatg aatgtggtct ctttctcaaa atatcttata gcactgtaat cacccttctt 5820

ttccttgtga tgtaaaatga cagtgcctat gtgctgagat gaggtgaggt ggatgacata 5880

ggcattatga cctggcgtta ggctactatt gacctgagaa tccatcaaac ttatgaattg 5940

tttatttctg gaattttcca tttaatattt ttgggccatg gtttacctca ggtaactgaa 6000

accacacaaa gtaaaattgc agaaaaggag ggactactat aataagaaga gaaggaagga 6060

gacaggaagg gcataaggca gacaggaagt ggaggggaaa gataggaagt gcaggaagga 6120

gacaggaagt ggaggggaaa gacaggaagt gcatgaggga gacaggaagt ggatggggaa 6180

agacagaaag tacatgaggg agacaggaag tgctggggaa aagacaggaag tgcatgaggg 6240

ggacaggaag tgcatgaggg agacaggaag tgcatgggga aaattggcag ggattatctt 6300

gaaaagacag gaagtgctcc agaactagat acttaggcat ccagggtaga gtggccccac 6360

aggctggagg gaagacaggg attcttgaga gactggagac caagaagaga ccctaacctc 6420

tgactcatcg ccatcctcca ggaaaaccgg gaggtggggag actggcgcaa gaacatcgat 6480

gcactgagtg gaatggaggg ccgcaagaaa aagtttgaga gctgagcctt cctgcctact 6540

gcccctgccc tgaggagggc cctgaggaat aaagcttctc tctgagctga aa 6592

<210>4

<211>26875

<212> DNA

<213> cTnI gene

<400>4

attttaaagc cgttagggga tgacctttga cagtccgcat cacccctctc acacaacgag 60

cgcctgttca aggttcttga ctggaagttc taccttgtat ctggcctcct gtagcagttt 120

cagtccattc cctgtgagga gggtgtgcca catggctttg ggggtcatgg agaagacca 180

ccttgcagat gtcctcactg gggctggcag agccggcaac ctgcctaagg ctgctcagtc 240

cattagagc cagtagcctg gaagatgtct ttaccccccag catcagttca agtggagcag 300

cacataactc ttgccctctg ccttccaaga ttctggtgct gagacttatg gagtgtcttg 360

gaggttgcct tctgcccccc aaccctgctc ccagctggcc ctcccaggcc tgggttgctg 420

gcctctgctt tatcaggatt ctcaagaggg acagctggtt tatgttgcat gactgttccc 480

tgcatatctg ctctggtttt aaatagctta tctgagcagc tggaggacca catgggctta 540

tatggcgtgg ggtacatgtt cctgtagcct tgtccctggc acctgccaaa atagcagcca 600

acacccccca cccccaccgc catccccctg ccccacccgt cccctgtcgc aattcctcc 660

ctccgcaggg ctggctcacc aggccccagc ccacatgcct gcttaaagcc ctctccatcc 720

tctgcctcac ccagtccccg ctgagactga gcagacgcct ccaggatctg tcggcagctg 780

ctgttctgag gtaaggctcg ggcagggctc tggggaagag gagagcagag aatggacggg 840

gagatgtgag ggtcttgggc cctggcatat ttaccccagag tctgcctgtg tccgcagaag 900

tccatggccc ctcctggtgg aggccacact tcagaggaca ggttgccagg tctgggctcc 960

aagattggta caatagagca gagagaaaga gagaatccca tggtagccat tggagactag 1020

ggttggagtc ctggacctgc cactaaagat gtagagttta aactcttgga tgagtcactt 1080

tccctcccca ggcctcagtt ttctcatctg tagagtgggc taaatatttg ctaggcctct 1140

gcctgctcta aatacaatga ggctctaacc aacagagact cccctaaatt gcttgttcca 1200

ggccgagggt cacctatacc tttccagtca ggtccttctt gagtcccagg gagcagagcg 1260

agactgcagc cctgtcagtc aagtgagtct cctggtcata ttctcaaggc taggaactca 1320

gaaactgtcc tggaagtact gagaacgacc agatgtgcag ggtcctctgc agcgcctctc 1380

caggccaacg cgggaattcc atttgatgcc gctgactcag tgccattccc tcccagagct 1440

gagctgagct cccagaccct gagctgagct gcacttgggg ggcagatggg gaaaggcccc 1500

actgggagaa cctgtccttg ggtgtgagcc cagctggttg gcattaaagg gtttaacctg 1560

ggcccccgta agtgtaaccc ttgtcccatc ccaccccaac acccccatctt gggacaagcc 1620

ttgctctgtg cccaggagtg gtagagaaag attcccttgg tttctagttt ctcctgggca 1680

ggctgaggac tattcaggga gggggttgcc tggtgtgtaa ggaatttcag agcacattcc 1740

tggcctgctg tagccgggaa gctacctgcc ctcccaaatt ccttttctga cccatgtctt 1800

agagggcgaa tcttccttct ctctgcaaaa gtggaatctg agaggatcag gttgctggct 1860

gcctcctatt tctgaccccc acttccactc ttcctgccct ctccccatcc ccttacccccc 1920

gctctcctgg tgggctaaca gctgttggct taggaaaatc ccattcgagt gctggggaca 1980

aggactgagg ggtggggcct gtgtttctca gaaactcgga gactgtttct acagggacag 2040

aactcaaggc tctcctgctt tgtagttggt taactgaagt cagagaggct aggtgactta 2100

aataaggcca cacagcaaaa tctgggtttt aagccccagat tttttgaccc caaggtcagt 2160

atgccttatg ccttctattg tctttgcttt actccccaag aaaggtgacc ctgtgaacgt 2220

gttaggcttc caacacgtgt tggctatgaa tgcattaggc tagaaagctg cctgtggaag 2280

tgggtgggac ttgggtgtga ggacacaggt gtttcaggac attctggttc cttgccctgc 2340

caggttcttg actctctccc attctgcagc ccccagtctc agccccacctc tttcccctgc 2400

atcaggtcct gcctttgagc cactggcatg gtggtgtcct ctgggctggc tgaaagccct 2460

cctccatcct gcatgtctgc atgctatttt tgggagctgg atggtgcccc caggaacagg 2520

gagaggagtc caagctgctc tgtcctcctg tgaggcccca gcaccatgcc cagtggctct 2580

gaactggtgg aagggcagag taggagga gagagaaagc tagaagctat gcgcccaggc 2640

tggccatgca cggttcctct gagaattttc tccctcccac tgagggacaa gcttctcctg 2700

gggctcaggt aggagctcct caagttctca agtcactagt tctggccgct cccctggggc 2760

tttgggtaac aattgtcagc cccacccatc ctagggttag gaaggctatt ctctttggcc 2820

actctcatcc taagacctat ttggagaacc tctggggttt gagtcttttt ttcagcagaa 2880

tgaggcttga tcccgcatta tagcacctcg cacatttgat gtctcttctt ctcacccact 2940

caccccaccc tgggggttgg ggcaaaaaag tggctcagag ctgcggttca gagctccttg 3000

taaacaaggc tcctccctca ctgtcctcac cctgctccag cagagggagc agcggaagga 3060

ccactctgct gcagccatgc ttgtttctaa cccagcagaa ctggacataa tgggaacagg 3120

gtctgaagac aatcaatcca ggctgcagtg ggtgctgagt ctggggaagc ctccacctgg 3180

aggggcagct gggcagtggc agctcccttg gaatggctca gcctctggac atcaccccac 3240

ccaaccagag ccctggctct tgctggatgt ccacagatga gtgcctggga ttggtctcag 3300

ccactatggg ggggatgtgc agggaggt gatgaggggag tgagcaggac tgtctatgtg 3360

cctctgtcct catcctgagg cttgggtctg aaattggtgc tgcagcactg gcacgcgtga 3420

gtggggcagg aggtgggcag caggggagga gcattggagg gtggtcctga gcatcatttg 3480

gattcttctc ccgcctcact ttagcctcac tctcacataa ccctgtccat tctctccctt 3540

ggaagccaac atcaggaagg ctgtggagct ggcatattgc actataacag aggctgctgt 3600

tcacgtttca aacaacgtga ttgtgtccct ggagcagtgc actgtcatgg ccccggaggg 3660

ttgggtcata gtagttatta gctgaatgcc tacacttcac actacctctg ttcctcacac 3720

ctacccaaga agagcagtaa tttcacagca aagagagctg agatgtgggg aggttaagtg 3780

accttgccaa ggtcgcaggc aaacgcatgc tggagccagc attcaaaacg agattagtct 3840

cacactcaag tgcttgctct ctgccccctc cctgtgatta cacgcgcact gtaacacctg 3900

acgttgccta atgccttccc cttttgttaa tgtcaactgg agcctcgtaa agaccctgag 3960

tagccaggat gtgtacactt tatctccatg agacagagaa aggactgagg tttggagatg 4020

ttaaaaagac tcgcctaaac accctcggca agctaatgga agaggatctg aatctagatt 4080

atctgttccc agtgcagtga gaagatgatc tcagaatgag ttccctgggg gaacattttt 4140

ttaatcggaa ttttcattga cataattaaa gactagtaca cagttgtctt taataacgag 4200

cgtttacaca cggtgcctag tttctcccga gggcaacatt tgcaaaatta gtactatatc 4260

aaaaccaaga ttggacatca atacaatcaa tctatgatct tattcagatt tgcccacaat 4320

tgcaccatgt gtatgtgaaa ctctaaacaa ctttaacctg tggaggttca cgtatccatc 4380

accacagtct acatactgaa aagatcccaa accacaagga tcccccctgt gtgctttgat 4440

ttatttattt agagacaagt tctcactcat ttgtccaggc cagtctcttg gggtgcccaa 4500

agaccttttg tccaggtctc acttttgtcc agcacacatg gaatgggtgc ggtggctcac 4560

gcctgtaatc tcagcacttc gagaggctga ggcaggagga tcccttgagc ctgggagttc 4620

gaaacagcct gggcaatgtg gcaaaatccc agctttacta aaaatacaaa aattagtcgg 4680

gcatggtggt gcacaactat agtcccagca actcaggagg ctgaggtggt agaatcgctt 4740

gagcccagaa ggttgaggct gcagtgaggc tttgtgcagt gaagatttgt acagtgctga 4800

acagatcctg tggagtgggg gtgggcagag tggatggaaa ggttgagaaa ctgcagacat 4860

tgtcttgagt gtcacaaatg gaaatgcagc atcccaggca gtattgcctg ttcaggggca 4920

aggaacgaag tggacatcag caggtggcct tgctgccatg tgggtgtcac tatctccccc 4980

agcaggggag aaaacaggct ttttgttgca ggtcacacag ctcatgaggg gtggaactag 5040

attcacccta ggcctcgctg atctctgtac aacgggggcc agagctcttc tgaggaaggc 5100

aggcttccct ttgtacctgc actgactttt ttctcctttt ggagggagag cagagaccat 5160

gtctgacata gaagaggtgg tggaagagta cgaggaggag tgagtatctg gagcatcttg 5220

cctgagtggg gtcctctccc gccgctgccc tgacacctgg tccaggagcc tcccagctgt 5280

ccctcggatt ctgggtagaa gtagctgtgt gtgttttggg caccccgagg agagatatag 5340

ccatttcctt tccactcctg tacccctggg ttgtagatcc caggcagagc acaggccgag 5400

agacttcggg aagcacagag gttcagctcc agggtcatca gtgtccttgg ccagtcttgg 5460

ctaggggcca tgaacagaga agctgagatg ttgggtctcc tgaaactgac tgaggaaggt 5520

tcagtgactc tccagaggca cagagctgat tatgcccccc aaacccccagc tcagaaccca 5580

gtgttatcac cctgtcttgg atccctactc tctcatgaca gtgccctgcc cacagtccta 5640

tcccagaggg aaaccagggg gtgggacaca gccaatccaa taactacaga ggatagtggt 5700

caccaggggag gagacagagg tataggcacg gctactgccc tcacagaact tactcttgtt 5760

gagatgttaa gacataaact gggagaagaa attagcaact gggggcctgg tgtggagggc 5820

gggctgttgg tagtgtagga gttctgagga gagggagatc aggggctgga atagtcaggg 5880

ggctgtgtgg agaaggcagg tcttacctgt gtagaatttg gaaagataag agatttggga 5940

gagggcagga ctttctagaa aggggtacag cttgggcaaa gatggaaaga gtaagtgagc 6000

agaacgctgg tgagaccaag ctggctgggg cggaggcttg cggaggatgg taggaggcag 6060

ccctgcatga gaacggcagg ccaggctagt gggtgtcatt gcaaggtggg cagggcagcg 6120

tggactccac taggcaacaa gggaaaagaa agggggatta tctttgggga aaggccagtg 6180

tgtgcatgtg tgtgcaggcg tgtgtgtttg catgtgcttg tgtgcgagct actgacagtg 6240

tttcctgttg ctctcaggga gcaggaaggt aagcgtaaac gtgtgtactc atttggatca 6300

aagacagcct ggttcgaaac tgacccacct cttctctctt ctcttgctgc ctggacttct 6360

cttctgagca gaagcagctg ttgaaggtat gtgcctcagg caggtggaga gccagaaagg 6420

ggcaggctca gcaatgtgga aatcctggga gccctgggaa agcagctcat ctgtccctgc 6480

tcctcactct cagctgagca agatcctcca acaacagtgc cttctaaaat gggaatcagt 6540

cctacctggg ttgctcgggt cttgaggcaa acgattaaa cctgtctctt ctgtctgcca 6600

tttgcttcaa acacactgat cacttttccc aatttttttt gcttcttctc ccttttatca 6660

tgggaaattt taaatgcaca taaaggtagc aataataata ataatggccc cccctgccca 6720

aggtacccat taccagcttc aacaatcatc atcttagggc cagtcttctt tcctctatac 6780

tccaccttcc accagttatttggagcaga catccaaaat tgttcccagg caccatgtca 6840

gttcacgctt caagatttcg ctacaccgta tctctaagag ataagcacta cttaaaaacc 6900

ataaccacaa tgccatgata tgccccaaaa gattccattt agtaatttct caaatgcgat 6960

tgtgagtatg aaattgctca gtgatctgag acgtcttatc tcggcatgaa gatttgtcat 7020

gattctgagc ttgcagaggc ccctcttgct gctcctatgc tgatcggtta acccatcgcc 7080

ctcctcagct gtgggtccca catctccctg aaagaacgcc ctcaccttcc cttcttcctg 7140

agagcttggc acatggaaat gactcagagg cttcactcca gtcctccccc tgcaccccag 7200

gcaggtggac cctgcctttt agtggctcag gtatcacttc agcatgtggc atgtagatat 7260

taggagaagg gtctctatga aggtccaagg ggggagccag gtggccccgc ccttgctccc 7320

catggctcct agggaaaatg gggaccctcc tgcacattta gggctgttcc agggctgtct 7380

ggccaaagag gagacagaag agaaattcac aggagaaact gaaagaggcc acaggcgtct 7440

tttttggttt ttaaggaaca ccagatactt cagaagctat cagaaatata tttttgaata 7500

aaaaatctaa ctggctccaa attctgctgt gttccctgaa cagactcttg atatgatatc 7560

ccttcctttg ttttggagga ttttccactt ccttataaca ggtttagggt ggtggagttt 7620

gagacataga tctctggatt tgggaccaat ctgcctgcca cacacagtag aatttggctg 7680

tgagttagca gagtcattct ccactgtgct ggggcctgag ccgtgtagca tctgccttgc 7740

ttgccatgaa tagacgagac ttgcctccca cacagttgag gatctggctg caaatgatca 7800

agagagagca acgtctagat gggtgtccac taagcctgca ggatagataa tgccaggtgt 7860

ccttcttcca ggagaggaat tgttccactt gctggaactg tttttttggg cagatatttg 7920

tacgtagaac aactgttgta gatgcagcct ctttccttag atttataatg ctggaccctt 7980

cttagatggg gcagcatcct cccaacaggc cgggcaaatc tcccctgggg ccaggctctc 8040

caatgaggct ttctaaagtg tgactcctgg aaactggaca tagtttgcag gatacccaag 8100

agcctcattc agagcaggaa catgggcagg ctcgttgagg gcaggaaatt tgatggaaga 8160

gaattgtccc ccactctctg ctctttaggg gcctctgtct tagtaagaag gaggccaagg 8220

ctctgtcgtc tagaggcatt gcagttacca cagatcgcat gctcctcttg tgccaggcac 8280

ctgctgggcg ctctatgcac gctaggtcat tcagtcctca ccacggctga gaagtgagta 8340

gaatgattcc cattttactg atgaggcacc tgaggctcag ggaagtaact gagagttgcc 8400

cctgttgctg gaaagtagtg gcttgggaat tggaatgccg gcctaactcc aaagcccaca 8460

ctcttaaaac cactgcgctg ggtggctgct cctgccgcgg gctctctgct cccagactaa 8520

cctgtctcgc ttttcccctc cgctgcggcc actccctgaa cctcagaaga ggaggactgg 8580

agagaggacg aagacggtag tacagccttt ccttctgtgg tgctttctgc tgcctgctgt 8640

cccaagtgca gcctccttgt ccaggggccc tgttctgggg gctggggggt gtgagtaggc 8700

ggcagggacg gagtgggtca gtcgtttcct cccctgcctc ccaggggcca ggatcacagt 8760

tccaagctga tcatcatgcc tgaacttcag ggccactggc tcctcttcct ggttcctgga 8820

tccttcccct gggttgggtg gtgggaggga ccaaggctag ctggtctccc aagctgctcc 8880

aaatgtggcc atagaggggga cctggcacct ggaagacagc agcgccaccc tgcccccgcg 8940

cacaccccag tgttccatgc tgggccctcc ttaggagctg gacagcacca agcagggtgg 9000

ccaggtgttg gttggggggt ctggggacag agtcctctgg agagcagcca gggagactgg 9060

aaatagccag agcaggaagg acatgacgtc agccttcaga tgcgccctgc tgatggggag 9120

cacaggacca aggcaaggga gtgagaccag ggcttaattt tagaaagtgc gttctgacag 9180

ctatatctca agatgcagta ggtgggttgg ggaggcagta agctccccat ccttgggatg 9240

ttgcatggcc actgggtggg ctggggtata ttatatagag aaatgacacg tcagatgagg 9300

tggcagaggt gtgaacaaga tggcctaaag gtagacccca caatggccat gtccttcctc 9360

gctagccctc tgctttcaca ctttctagaa ggatcatttt gaggtaagac tcctgcaggg 9420

atcaaataaa aggcatctcc aggcttcaga gaggactcag acctgggaag ggaggacaga 9480

gggaatgaag gccaaccctt ctcccacgat ggtgtcttcc tcctgtctga ggtttttctt 9540

ctccctctgc caagaggcac ccacttccca gtgcatcagt gaacaggcat cacagaacac 9600

ttcctacagg cacagcccag gccaggcggc atcctgtctc ccttatctgt ttgtctgtct 9660

gtctacacca agtgcctgag tgagatgggg gccacatcct gccaaacaaa tagatgaaaa 9720

tagaaaaatc ctgcccactt tctggatagt acgaagatcc agatattgag tgtgccatcc 9780

agccacacag tcgtactctt ttggtttctt tgagatgtgg attgagggtg tctagagtca 9840

ctaggttcag gacaggagct tctggacacc tcggcttaag cagagcatgt tctgtggtgc 9900

cagacacggg tgaaacttct gggctggaaa gttgggaatg aggtggagtc ccatcatcag 9960

ggagtttgcc gggatgagtg gggctaagct gggtggcggg aggtagccga cagtcaaaac 10020

caagaatggc agccccccag cacgttgttg cccccaaatt catctggacc tagacgtctt 10080

gacccccatc cccactcccc gctgcccatt ctctgctctg ggttctgcct gatagcatga 10140

gtggggcctg cttcttcact ctgtctgctt cacttctcca gagcaggagg aggcagcgga 10200

agaggatgct gaagcagagg ctgagaccga ggagaccagg gcagaaggta agtctctggt 10260

tgagggtgca ggggcctcag ggggtggcga gagcccctgg agccattcct gaggcagtaa 10320

gaaaccccccc agaacaggct ttcccatgtg cacctgactc tcctattgac taagattcaa 10380

ccattgattg cccaatcaac aaacgttgac ccagcgcttc tcttgtgtca ggcagtcttg 10440

ggtccagggg aatgtgtgtg tgagagacag ctctgtaacc ccaaaaacca acagagtgca 10500

tcttggctag ggcttatctg tggggaaggc tcttgctggt ggcggagggg gggacttgag 10560

ctcagaacaa ggaccttgat tgatttctcg tgtttcattt agaagatgaa gaagaagagg 10620

aagcaaagga ggctgaaggt aaggatatca ggagagaggt acaagggagg caaaggaacc 10680

cctcacagca gctttggagg agaatgccca cagtgctctt tccagggaag tggatgtgca 10740

ctgagaggt ttgggagagg agaggacaga acttggtggc acttccaaag ttttagatgt 10800

tgagctttca actttgcccc ttctccccaga gaagttgcat tggtggcacc cagtgggaga 10860

ggggaccatt gtccagcccc ctggtctgga cagagcaatg ggacccagaa gtcatatcca 10920

cacatgaagc tcctagagct taggtgaaat cacttgtagg atgtggtgtc cgtaggatga 10980

aatcccaatg catagatggt catggtgggt cattgctgca gtcatcgtca tgctgcagcc 11040

ttcaccaggg ttctaagtgg tcccagtcac ttctccaaag gggaggggac aaccagctgg 11100

ggcttggaat catgtcatca gcttctgccc tgtggccacc gggaagtcca aggggtaaca 11160

aagaaggcaa agacctggga ggggattcta gggccaggac ccttggggac ctttccaaag 11220

gccttgtcac tgtgaagcac aagagaagca aaattgcttt gcccagaact ggtgagtgag 11280

agaaggcttt gtgtgaagtt ccacactgcg tgagtgtgtg ccctgcagaa ctctatggga 11340

gggggagtgt ggcatctgca gcccccgctg ccttcattct cctggatgct tctggaatca 11400

gggctcccca gtgccggggag ggactcactg tgcagatggg gaaatggaaa tccacaggga 11460

tctagctcat cgagggcgga gttgggacca cttctggggc ccctgaccca tggctgtgtg 11520

cttctatctc ctccagatgg cccaatggag gagtccaaac caaagcccag gtgagtgggg 11580

aggcacctgg gtagagccgg gagcagaggc tcagggagag aggatgatga tggaacaggg 11640

tgcagtatgg tgcagtggac acgggctggg ggtttggtgg gcacagagca ccacagtaca 11700

gttgtacaag ttgtgtactg cacaagcgtc tcttcctaag ggagtgagtg agggctgaat 11760

tccacccaac actccatcac caatcgtgca ccctagcagg gatggggcta tgttcttcta 11820

gaggaagggc cctgaacgga cactctcctc tctcacagta tggccagaac accctcctcc 11880

atggccgagg tgggaccttg ggattaaggg aagcaaattg tggggagcca aacaatgaaa 11940

ccgtgccagc ataggcatgg cggcttcaga gagctgagag agggcaggac ggcttggtgc 12000

cacacacgtg aagccccacct gaaggtggtg tgtgctgggg ggggctgggg gataggggagc 12060

atttcccttt ttatgatgcc aaccctgcca gcagaggttg acccatggca cctctgccca 12120

ctcacaggct cttcccattt cagtggggcc ttctgaaacc agcccaagac atcccagatc 12180

catttatttg cccagtgcct tcccccttcc tggcctcttg tgcactgcgg tatctcattt 12240

cagctccaca cacctgaggg cagatgggga catgaagaat cagagaaccc ctgggtctca 12300

gtatgtgttg ggaccaggac caaactgcaa gactcccagg ttcatggcca gtttctgcgg 12360

ggtccaccag gggatttgaa gtccaagaag catggcatct tgtcacagct tctttgattc 12420

caagttgtgt ggctttgagc aagtcattct acctccctga gcctcagttc tcctaagtga 12480

gattaacggt accccacttca tgggcatatc atgagcatga agtgaaatag catatgtgaa 12540

agagctttgt gtattcccag gtgctgagca aggctgagga ctgccattgt tgacgtcagt 12600

gttactatca ttgctgtggt tggtcggggc agtgctggaa gattctctag gaaggatcag 12660

gccctgcctg tcctggacac cctcagtccc tgggtccaga atggggctga tgctgactat 12720

tcctctctcc aacaggtcgt tcatgcccaa cttggtgcct cccaagatcc ccgatggaga 12780

gagagtggac tttgatgtaa gcggtggctg tgggttgagt aggcctgggc tggggatagt 12840

cccaagggcc ctcagtgaca gagattttgt gaccttgggc tggggtagag cataggtggg 12900

atggggtgca gcacagccag gctgggtgcc catccatgag cgtggtcctg ggttcaggcc 12960

acagttaccc cttccctggc caggcgccat gccctgccag aggtcttttg cactgcgttg 13020

ggggtgtcta gcccacccat ctctcctctg gactctttgg agtggcagcc tctgagccgc 13080

agcggtccac ccacaggaca tccaccggaa gcgcatggag aaggacctga atgagttgca 13140

ggcgctgatc gaggctcact ttgagaacag gaagaaagag gaggaggagc tcgtttctct 13200

caaagacagg atcgtaggtg tccagtctgc ccacctaaca ttctctgatg gcccagtctg 13260

tcctccatcc cttttgtgag gccccacctc ctccagacag ccttcctcgg cgcctcaaac 13320

ccagctaggt gtgaggtctc cttcagaggc cctccagcat tctgcacagc tcctgcagcc 13380

ctgactggtt tctcgggtcc ctgccttccc tccccatcct actggatgct ccttgactgc 13440

cagagctgag tctgatctgt ttcaggcctc ccccagggcc taattcaggc ccgggctgtg 13500

gactgattgg tcacaggcct gctgcgccac ccctgccctt gcctcctctt cccccgcaga 13560

tgaaggggag tgtgacctgc agaatcccac atccttctcc tgcagagcgt ggaggctgct 13620

ttgattgctt ttgtttctcc ccatcaaact tgccttccca cagacatgct gctctggctg 13680

gagcaatgac ttggccatct atccaagcat atggcacaga gcagctgcca gggcagggtg 13740

ggatggaggt gggggggcag cccagaagga gcctgcctga tgtctcccag ttgtggaggg 13800

cagtctcacc atgaggaatg ttggaggtgg gtcactgcag gtaccctggt gtgcacgtcc 13860

gtggagctgg ttgaaagtgg aggcccttgg aggccgggca ccattgcttc aagactctgg 13920

aaggaaccct cccccaggaa ttcccttgcc accccatgca ggtttctgta cctgcgatgt 13980

caccttctcc ctatgcacac ctgggtgggc tttgcctctt ctccttagga gaaacgtcgg 14040

gcagagcggg ccgagcagca gcgcatccgg aatgagcggg agaaggagcg gcagaaccgc 14100

ctggctgtga gtgaccccta gcccaggccc atggaggccc cccatgctcc cagctcctgc 14160

tgggccccca ggcccctctc tattcatcaa catgaatgaa atcagtcaag aagacaatatt 14220

ggccgggtgc ggtggctcac gcctgtaatc ccagcacttt gggaggccaa ggcgggtaga 14280

tcacttgagg ttgggagttc gagaccatcc tgaccaacgg agaaaccctg tctctactaa 14340

aaatacaaaa ttagccaggc gtggtggcac atgccggtca tcctagctac tcaggaggct 14400

gaggcaggag aatcgcttga acccaggagg caaggttgca gtgagccgag gtcatgccat 14460

tgcactctag cctgggcaac aagagtgaaa ctccaactca aaaaaaaaaa aaaaggaaaa 14520

aaaagagaca atatcaatat caacctagga agtgctgggc attgtcttgg ctctgataat 14580

acatcagcaa acaagacaga aatgcctgcg tccttatgaa cgtgttattt tataagactc 14640

tccagccctc atcccttccc cagtgcagac ttcgtgggaa ccgaaggccc cgtagctctc 14700

ttcctgtgca gctgctgaag gcatttccag agcaacaggt cttccagggg cactcccagc 14760

ttaaggggat gacgagacct cacatttggg aatcaggacc tcattcaatc acgcagtcat 14820

ttctcagtca gtggactcct gtgggccagc actgtgctgg gagctaccct ctcagaaagc 14880

tccttgctga gcggagagaa agctgaactc acccataaag accacaagct tcagcccaga 14940

atcagggttt ccaatccttt cccctaattt gctttcttcc tccctgctgt aaatcaggaa 15000

gagagggctc gacgagagga ggaggagaac aggaggaagg ctgaggatga ggcccggaag 15060

aagaaggctt tgtccaacat gatgcatttt gggggttaca tccagaaggt aggtagggag 15120

cagcaggggt tgccaggga tcctagtata gccctgagga atgaggtgtc cactgcagca 15180

ggtagacttt aggtcaggtc ccaggaagag attcccagct gctgtgggtg gaagactgag 15240

gtctcctcat gcccagccct gagctggttt tcagcctgac cgcggcttgc actaggaggg 15300

gcttgctggg gtagggggct acaggcagag cctggggcac caatggtgcc ccccatgcct 15360

ggctccagca gtgctcagga gcccccaagg aggctgggtg cagctggaag agccttgtgc 15420

ctagagtcag agctctagtt ggagaccag ctctgccaca tacagttgtg tgaccttttg 15480

caacctctga gcctcggttt cctcaactgg cactcatatt gcctgccttg atgctttgag 15540

aagctcaaat gagctaccat ggaaagtgtc tatgtgccac acagatgtga ggttgtttgt 15600

tcctggaaac tcactctcga tccctaactt ttgtgcccca ttcctcctag gtcctccatt 15660

ctcgagggag gagggatatg tgaaattgat tttctctagg tagtgaagga atcaaaacat 15720

tttcttagat ttccagctta gtccctagct caggactagg ccaccccctg cctctgcccc 15780

catggtgacc acaccagcca agactttggc atcaaagtct tcatgctacc gaacagcccc 15840

tgctgtaacc ctcagaccca cgtcctagta aacccggctg actacagcga gtatctgaat 15900

cttctcttcc atgtctctcc ttgcctttgt cttcctgcct tctctctgtg tctctgttgc 15960

cgtcttggtc tttttctatg ggcctctctc tcatggtttc ccggtttagc aggcccaggt 16020

gggtaccaaa aacctcaagc cttcttcctc cccatagttt tggagagctc cctgggaggg 16080

gaaggcgcct tgctggaggg gaggagtgaa gagattgggc tggctgggga gggggaagc 16140

tggtgagagg tgaaagagat tagttaggca aagaaggaag agactgacgt gatccttctt 16200

gcccctacct cagtttccct agggaataca tgtagttggt agttgggggg tgtggcagga 16260

agaagagcat aagaacctgg ccctcctgca ggccttgctc cacccccagg gggtttgggg 16320

agggttaggg gacagggagg gggcaatctg gccagtttac tctgcttccc acaccctcca 16380

gacagagcgg aaaagtggga agaggcagac tgagcgggaa aagaagaaga agattctggc 16440

tgagaggagg aaggtgctgg ccattgacca cctgaatgaa gatcagctga ggtgggggct 16500

gggaggaatg gtcctggggc actgcccgca tctgctctgg gaggggtcgg gccaagggca 16560

cttcttgctg tgagccacca gagggcagga ctgcctcaca tattttccca tggcaagttt 16620

ggtgggacca gagctgggcc ccagcaggtc gggccaactc ctttctgggc tttgctgatg 16680

gggaaagaga actgagcagg tgccccaccc ctctgagctt cacctaccca cctgcacacg 16740

ggtgtcagga caggggattg aactagatgc tctcctgcgc cagaagggct tctcctgctc 16800

cccgcttgtt cttggaactt tgccctggca gtccctggtg tccgttcctg agcactgggt 16860

gctgccgtct ggtccccagg ctgggtcagg cccaggggggt gggctggggt acccttggca 16920

ggcctggagg tgggaaggag ggctgggatg ctgggggctg ggccgggacc aggacggagg 16980

cccaggaggg ccctttctta ctggacctcc ccattctcac ccttgcccgt gcagggagaa 17040

ggccaaggag ctgtggcaga cgatctataa cttggaggca gagaagttcg acctgcagga 17100

gaagttcaag cagcagaaat atgaggtggg ccgccatgct gtccccgccc agcatctcca 17160

tctcacactc ctcctggttc actgggtccg gtaatactgc tgcaggtccc tgggtccctg 17220

tccctattcc ccaacagccc ccttcagctc ctgcatctgc ccctgctgcc tggccttcag 17280

cagtgttcca tgtccaccgc attgacgact gcttgctgga agtgtctgag agctcgctgg 17340

ggctgagcag agacactttc ctggtgttcc aaccctgggg gtctccaaca cgttgaggca 17400

gcagctcagt gatctgagct ggttacaagg accaggatgc accaagccag gacccccagt 17460

ggaaggggag tgctgccaac agagggtgc ttctccccac atacacccaa ggtcctggtg 17520

tgggcacaat taggctgagc ctcaagctca cagtctttcg gagtcctatg tgcactaatg 17580

agggtcttag gtgaacagac accaggcaag gaaatggctt agaggacact gatgctgcat 17640

accggagctt agacctgggc gccagtccctt ccttaccccac cacccccgag ccccggtccc 17700

agggcctctt tgctacctaa gggaagaatt cagcttcccc tggaaggtct cctttgctgc 17760

tcctgccaaa ccactcctcc ctgggcaaga agcccctcgc tgggcaggct caggctgcag 17820

tggctacagg gtactcccac ctcccaatac ggagagaggc tgtattgcct ggtgacagtg 17880

gcatggactt tggagccata atgcctgggt tgaatttcta cctgtgcccc tcactggctg 17940

tgtgacattg ggtgagttag tccactgttc cttgcctcca tttccacagg taacactaca 18000

atatacttca gagggtgatt gtgagggtta cagagataat actaattgtt attattgcta 18060

tagtgttcca accactgttc caagcatgtc ccatgtatta acttactatg ccctcatagc 18120

agccctatgg gttcatatct gggaaggtgc tcaggacaga gcctggcacc cactaaatgc 18180

tcagcaggtg tcagccattg ttatggcctc tctagtcctg tgccttccac ttttttctct 18240

ttttttggtt ccaacactgaa ctctgcaccg gccaacagga cacagatttg ccaaactttg 18300

gggcagcacc ctgcaggggt ggtgcatggg gatgctactg ctcaaagggc acagcttccg 18360

ggatggtggg cagctgggca ggggtgcccc agggggtct ggggctgggc tgctaggagg 18420

gctccatgac acagcctcca gctttgtgcc cagctctcag aggcccttct tatgggactc 18480

tcatatcctg aacctattat ggccctggga ccccacagtg ggaggcccat gaggcatcct 18540

ggaagcttct ccttggcttc tgcctgtggt acacgggccc ctcctgaccc ttaactatcc 18600

taacccctcc tactcttcca tgctcctcct tctcctcctg cactgctgca ctcagccccc 18660

ttctccccat ccccaggcca cctgggacct gagccagtca gctccagcgt tgctctttgt 18720

ccttcccact tttcttgcag atcaatgttc tccgaaacag gatcaacgat aaccagaaag 18780

tgtaagtgtc tgaggtcatt ctcgcctagc cctccccctg ccccctcctt ccctaccttc 18840

cagctatccc attcctcctt ggagggcccc aggctcccag cccttcctcc tctgcttcag 18900

cccacaggtt tccttctctg gctcctccgg ggcctcgtgc ttctctccct gtggcctctg 18960

cttgctggtg ggggtgaaat gtggggcgga gaagcttccc tccagcctca ggctgctgtc 19020

ctattctgcc cctggaggca gctccagcct gctcctctcc cctttggcac cccagtccta 19080

ccccagccgc atggtgacct actaccctgc ctgtgtctcc atgtcactgc gtcctgcttc 19140

ccctgcagct ccaagacccg cgggaaggct aaagtcaccg ggcgctggaa atagagcctg 19200

gcctccttca ccaaagatct gctcctcgct cgcacctgcc tccggcctgc actcccccag 19260

ttcccgggcc ctcctgggca ccccaggcag ctcctgtttg gaaatgggga gctggcctag 19320

gtgggagcca ccactcctgc ctgcccccac accactcca caccagtaat aaaaagccac 19380

cacacactga ctggcatttc tgtcacgtcc ccttcagaga gagagagaaa ctgaggcgag 19440

accctgctct cacaggtccc cggtgtctgc caagctctaa ggagctttcc cctgtctggt 19500

ctccctgctt gcctctagct ggggagaggg tggagggccc agggtgcagc ccaggaggca 19560

ggagcagtgg aaagggcttc agatcattgc tcctgggagt caccaaaaca gcacatggga 19620

aaagtgctgt tgagggggac agagtgttcc ctgcatgagt cactcactgg agaacaatgc 19680

aggactcttc ggatggttcc cgttctcttt ctcctcctag ccagggccct aggaaccagg 19740

tgtctgaggg acaggagct cttgaggaag gaaggaaggc cagttttatt gggcagctat 19800

tcaggcctga gttgttgaga ggtgtgggtt agtctgggtg cctcagtttc cccctcccat 19860

gatctgcaca tcctcgaggg cttaagtcac agggtcctga gctgaagagg acagcttggt 19920

ggcatggatt tcagttggca gaaagacaag gaaacgatga gagggcacca tcctcctctc 19980

cagaggagag cctgagacga cacattctct tgaccctcca ttcctttgtc aggcgtggaa 20040

tttcccccaa ttcctgattc cttcccttgg ctccccactt gccatccatc agcaagtgcc 20100

tcatgagctg tgtctggaaa ggcttcctgt ggtctcccag ccagagcaaa tccatttctg 20160

cttgctagag catgtgctgt cacagctatg ccatccaatg ccacactctg ttagacgggt 20220

ctcttcctgc aggcgtggct ctcatgctcc caggagacaa tgcagacatg ttatcctgtg 20280

ctgtatgcca agcactggga tccatgtagt gagagatgaa gaggaaatgg tccttgggcc 20340

acaggggaggg gcttagcctg gggtctcggc ctcatttctt attcattcat tcgttcagtc 20400

attcaaatac ctcctgagca ccaactatgt gctagacact gtgttgggtg tggggtgctg 20460

tgatttaagc cagcacatcg ctcatggagc tcacagagct cacatccagc agcagggaga 20520

cagcaagcag aggtcccagc cctgggtgac aggggcgggt ggggggtgcg atgctgggga 20580

gaggcatagc cacgcaccac acctggccct ggcaggtggg aaggctttct ggaggaaaga 20640

gtgtctaggc cgagtaccag agggtgagga gggggaagtc aaggggaaga ggttgggggt 20700

ggggctggga acggcacata gaaagtttga gaagcaagag ccagcatggt gtgtttgaag 20760

agatatagtt attagcagaa tccaagcatc ttctggtggg gacaagaagg ctccatcata 20820

aatgccccca aaatggacag ggaggcagga cgcaggcctc aggcccaagg agggctgtca 20880

ccacgggctc tgggagaccc actggcagct cctggcacgc accagcccta ctatctcagt 20940

ttgccggata ggacagctgg ggcagtgccc tagcccctct ccccataaag gggcactgca 21000

cctctacccc cacctgccgc cttggctcca ccgcctatgc ctcagtgtct ccttggtgca 21060

acaagaggtc atgatgggcc gtgtgtctgg ggccctgtct cctgctcccc ctctcccagc 21120

agggtccttg tgaccatttc tctagtacag cggtccccaa cctttttggc accaaggacc 21180

agtttcttgg aagctattt ttccatggac aggggtgggg tggggaagaa aagtagggggt 21240

gggtgggtgg gggtggtttc aggatgaaac tattccactt cagatcgtca ggcaatagat 21300

tctcgtaagg agtatgcacc ctagatccct cgcatgcaca gttcacaata ggtttcacac 21360

tcctatgaga atctaatgct gctggctgct gatctgacaa gaggtggagc tcaggccgtc 21420

atgctcgccc accccacagct cacctgctgt gcagcctggt tcataacagg ctatagactg 21480

gtagagtctg gtccacggtg cagaggttgg ggacccctgc tctagtagat gtggtctctc 21540

ttttaaaaat tgtgcccagg ccgagcgcgg tggctcatgc ctgtaatccc agcactttgc 21600

gaggccgagg caggcacatc atgaggtcag gagatcaagg ccatcctggg caacatggtg 21660

aaaccctgtc tttactgcac tccagcctgg caacagaacg agactccgtc tcaaaaaaaa 21720

aattagctgg gcatggtggc gcgtgcctgt agtagggctc gggaggctga ggcagggga 21780

tcgctcaaaa aaaaaaaaca actgtgccca gaggtgggca cagaagacac ccttgccctc 21840

cgactctgaa aagcccctaa gaccagatcc cacttaatct cttttctcct aaggtcacct 21900

ggtgttcagg aaagaacttc ggagtgtccc aggcgctgcc cccatggggg cctcacccag 21960

ttgaggtccc catcccatca tcttaagagg aacaccacag gttaacccaa gcgctgcctc 22020

taggcaggag atctgcagac acctggtctg catggaacct ggagcactaa tccataagtg 22080

gagcctggga aggggacctt gtcctatccc aggacagagg agaaagcaag tgagtgtcag 22140

ggtgcagggg cacacctggt ccatcagccc agtgcaggga gtcctgccaa tttcaaaagg 22200

caagggtggg ggcttcagac caagtggtcc tgtgctctcc ccataatatt gctagggaac 22260

agaaagatca tggatgtgtg cacagaacac actgagatag cttcacagat agggcctcca 22320

cacaccctcc tcagttcacc caccacttcc atcattcttg tgcttatgat ggtcctgaac 22380

caaacgttga aggtgcagca gtggccaaga gctgtcctgg gcctcctgac aactgaggag 22440

ctggatgcag cggggtgagt tgacgcccac ccagacactc tgaccccac tgtccccact 22500

gtcccctgca gggacctccc tctatgcctc tcagatccca tctcactgta ctttcgatcc 22560

atgacccatt gcctcatggg cctctcagca ccctgtgtac accaaatgct cagggaaaat 22620

tacttgcagg ctcttctcag ccctgagtca gaggtgaaga ctcttccgcc atcctgtcat 22680

cagattcgtt ctcctaacaa gctcgcatcc tgggcagagc ttgttcgtgt gatgcccact 22740

ttagaggtag agctgggaga gaacaaggtg ttcccccagg gctccttcgc ctgggaatcc 22800

agctcctcca gctggagccc cgatgctctg aagctagagc ctcactttct cccatcaaac 22860

tctggctctg ccccttccca gctctgggat tttggtcaaa ttccttgcct tctctgtgct 22920

gaagttcccc tatctaaaca atggggattg ttatgaaggc taaatgaaat aaacatgcag 22980

caactagcac agtgcttagt gcatagtgtg taatcagcac gagccactgc tactgccttt 23040

tggatgaaaa cacgttatta gcctttctca agggctcaag gtctgcagaa gaagagttca 23100

cccatgctgg gtctgggctg gctcctcctt gcctgtgtgg ttttagggaa cccttcaatg 23160

gctcctcttt cctcctggat tccacggcct ggtcttctcc ctcaccagat aggccaccag 23220

gggttggggg tgggtggggat ttgctgctac ggactgtccc tcctggcacg tggttcagcg 23280

tgtgccactg tctggatgga ggtgccagaa cttgttccca ggccaggctc cactccctac 23340

tctgcagctt gatttctggg aaaggtgggg cagggcctga gcttccaggg aacctgccct 23400

ggttggggga aatagctggg ctcagggata ggtctcaggg ccaggctctc taataggctc 23460

ctgttgggta tggggcaggc tgggggtgca ggggagacca ggctggaccc catgttccca 23520

cagtgtagcc acgggcactc acttgcttct ctgggcttgg cttctcctct gaaagtccag 23580

tctgccctgt catccctacc aggaggaaga ctaagtgtga atttggggaa aatgctgctg 23640

atctgggtca gctcgtcttc tccccctgca cacataccct tcatgtgttt ctcgctctgc 23700

gccccccacg caggcactgg gccagctact ttacatatgt tctctccagc caaccatcca 23760

cagtccccag atcacaccgc atgttgcaga tgaggaaact gaggtacaga gagtttaggc 23820

aacatgccaa gggccatca gttagtaagt ggcaaagctg ggatttgaac ccaggtctat 23880

gtgacgccag agccaggatt cttcctaaca tgtggatctt gtagattgag gagcatccca 23940

aatctcccca gattgcagtg cttgggccag aatcagagcc tcccgtgggc tgacttgggg 24000

tgagggcaca gggcaggagt ccaggagctt gaccctctct gctgccctcc agctgcccac 24060

ctgggacctg acaggaggag atgtgctatg tctagcacat ggaggcggcc tctggaggga 24120

acagtagagg tggggttgag tggggctcta cccagggtgc aagagaatca ttggccagcc 24180

tcctccttct tgggagtaat tataccccat acctgtgtcc cagagtgtt ggggggatct 24240

gtgggtcatg gaagcacctg tcattacctt gtctgtgggt tacccctcga ctctgcaaag 24300

atagtgagta aacctcaggg tgggcagaat cctgggtcta gatgccagca gggcctatct 24360

ggctgttgtt gggggtggca gaggtgcttt agggaccctc tgcagtctac ctccagcctt 24420

acttttcagc caaatggagc tagctacttt tgtttctcca gcatgtccta tgctctccca 24480

cctctgcatg gctttttctt gcccttcttt ctgtctctgc ctatccaact gccctccgcc 24540

ttcctcaggg cctcgatgaa gtccctgttt ctccaagaag cctttgggac tccctcctga 24600

gcacctgggg cttcctgagc accagcgatg cctctggtga tgccttaaga gccgcacatt 24660

gaaggcccat tcactctggg cctcctgtgg ttcacagaaa ccaggactgg gtgcccagcc 24720

cagacctccc tgcctggcac tgttcccagg gcttcagccc cacccacctg ctcatcccag 24780

gttctcagct cccaccccag catctacagg aatttaaatt gctctcaaat gaagactcag 24840

gagaaaaatg tttccatact tgtgcagagc gccctgacct ggggcggggg ttgggggaag 24900

tagccccagc tccgccctgt ggccctgccc tcagtgctgg ccttggccac actggccaca 24960

cctgcagcac gccccaatcc caaagcagtt ttccccgtaa agccaggcct gagcaatgca 25020

aacctgaaga agcctggggc taggctgggg ctgctcttca ggaacgcagt gggcggggtg 25080

ggagaggtgg tgtgattccc aggctgagag gtgctgagat tgtgctgggt ttgctaaatg 25140

tgggacagggg agacctgttt aggatgagct gcctcacccc gtcctctccc cacccatccg 25200

taccctgcca attcccagaa gatgatgcca gctaggaatg caggttgatg aagataaagg 25260

cagaaggtca cttcacttct acctgggggc aatagacaaa gcagtgtttc cctctgaact 25320

tcacacagag actcctctgg ccccacatct cccacctgtt caggcccagg tgatgtggtg 25380

agcagtggtt tctgctggga gacacttggc ataatttgta agccagctac atgataatac 25440

taacaaatga tatttgcctc catatattag gcacctatcg tgtgccatat actatgttac 25500

gtaatttatg tccactctct aattacagta acatgcaaaa ttaggtccta tataataact 25560

ccattttata cacaaggaaa ctgaggctca gagaagttag gtgactcatc caaggtcata 25620

cagggcagga tcctgattac tgtattttta ttacaatgtt aaaattattt gaatggtaca 25680

cttcctacca aatcaagtca ggcatctgat agaaacaaag aaaagcatat tttcaagaag 25740

tcccttctgt attttagcca acaaatattt gaataatttc tgattatata atgcaaagta 25800

cgagtgcatc gatccttatc atagaaaaac ttgtgtgaaa taatgttcaa atttgtctat 25860

tgcaaaaatt tgatcactca cctgggaaat ctggaattag gaatgtaaga tcttaccact 25920

gcctcttgct catataaagt ggcttgtgaa ttcacctagc cctcatccca ctcagttgcg 25980

gttggtggtt tcctcatctc tcccctacat ctagcctggg cacttcctgg gacctggggt 26040

cctgggttgt tcatccgtag atcccagtgg cctccatgca tgggaatgaa tacaggagaa 26100

aaatcattct caaattagat gatcttgttt aaggtttgat cttttaaata tcaaatatca 26160

gagagggaaa atatggcaaa ctaatcacat tagagttgca ttgtctttca cagttgtcaa 26220

agcactgtaa caagtccata ggatagtaac tttagaaaaa gaagaccttt gatcttggaa 26280

gggtcatacg tttacctcgc tcacatcata tcgatgcaag cattaaatct gcttgataac 26340

caatgtttct attagaaaat agggggtcact gaacacattgg ttcagtcatt gtgtaaatcc 26400

aacacatatt tattaaacac atactctatt ctaggtactg gctagttatc agggttatac 26460

ctgaggacag accaagactg tttcctacct tcataaacaa gaggagaaaa gtattaaaca 26520

gacattagat cactggatag atagatagat gatagataga tagagata gatagataga 26580

tagagata gatagatagg agatagagat aggtgacaga tggatagatg gatagataga 26640

tagcagatag aaaagagata ggtagataga tgggaggaag gtagagat agagataggt 26700

ggatacatac atgcatatat gcatagacac agacacatgt atgtacagat atagatatac 26760

atatgcagtg cctgggtctc aaattgtaaa aagagaaacc cagaggaaaa cagcagtgtg 26820

ctgtgaaaga gaacaggttg acataactta aacggagttt ggatgaggaa gcgtc 26875

Claims (10)

1. an isolating sudden change β-mhc gene or its fragment, its following position in the sequence of listing corresponding to Genbank accession number gi:M57965.1 (SEQ ID NO:1), contain the one or more sudden changes that are selected from down group: 10353G＞A, 10429A＞C, 10895A＞T, 11811A＞C, 13653A＞C, 14449T＞C, 14452G＞A, 14557G＞A.

2. an isolating sudden change cMYBPC gene or its fragment, its following position in the sequence of listing corresponding to Genbank accession number gi:Y10629 (SEQ ID NO:2), contain the one or more sudden changes that are selected from down group: 4826G＞A, 8697G＞A, 8887G＞A, 12101G＞A, 13261G＞A, 14262-14264de1AAG, 14364de1G, 18115-18116insGCAAG.

3. an isolating sudden change cTnT gene or its fragment, the one or more sudden changes that are selected from down group are contained: 4693C＞T, 4769G＞A in its following position in the sequence of listing corresponding to Genbank accession number gi:X90780 (SEQ ID NO:3).

4. each mutator gene or fragment among the claim 1-3 wherein also comprise extra sudden change or nucleotide polymorphisms.

By claim 1-4 each mutator gene or the protein of fragment coding.

6. nucleic acid, its can with each mutator gene or fragment or its complementary sequence specific hybrid among the claim 1-4, but can not with the nucleic acid hybridization of corresponding wild type gene or its complementary sequence.

7. a test kit wherein comprises the described nucleic acid of claim 6, and working instructions.

8. a test right requires the method for the sudden change that each limited among the 1-3, comprise that application rights requires 6 described nucleic acid as probe or the primer detection of biological product that imitate, perhaps Application Design becomes will obtain comprising the step of segmental primer to increasing this biological sample and detecting of this sudden change after amplification.

9. a microarray wherein comprises the defined nucleic acid of claim 6.

One kind the screening or the assessment medicine method, comprise that cell, tissue, organ or animal individual to carrying each described mutator gene among the claim 1-4 give one or more medicines, and assess the index of its one or more indication hypertrophic cardiomyopathy situations, with indication amelioration of disease situation.