CN112094822A - Infectious cDNA clone based on EV71 strain and application thereof - Google Patents

Infectious cDNA clone based on EV71 strain and application thereof Download PDF

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CN112094822A
CN112094822A CN201910474088.3A CN201910474088A CN112094822A CN 112094822 A CN112094822 A CN 112094822A CN 201910474088 A CN201910474088 A CN 201910474088A CN 112094822 A CN112094822 A CN 112094822A
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易志刚
宋志刚
张惠英
袁正宏
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Abstract

The invention belongs to the field of biological medicine, and provides a stable infectious cDNA clone based on an EV71 strain which is clinically separated, derivative clones containing various reporter genes, and various mutant clones constructed by taking the infectious cDNA clone as a female parent; and various recombinant viruses, subunit viral particles produced using these clones; and animal models established by infecting animals with various recombinant viruses produced by the clones; and the use of these viral or subunit viral particles for vaccine development and diagnostic reagents; and the use of the virus as a gene therapy vector or an expression vector. The invention provides a new tool and a new way for detection, prevention and immunization of EV71 virus infection, and provides possibility for gene therapy and vaccine development by using the EV71 strain infectious clone as a virus vector.

Description

基于EV71毒株的感染性cDNA克隆及其应用Infectious cDNA Cloning Based on EV71 Strain and Its Application

技术领域technical field

本发明属于生物医药领域,具体涉及基于一株临床分离的EV71毒株(js1)的感染性cDNA克隆的构建,以及利用此cDNA克隆及其衍生克隆产生的病毒及其带有报告基因的病毒及建立的动物模型在抗病毒药物研发,疫苗研发,病毒诊断方面的应用。The invention belongs to the field of biomedicine, and in particular relates to the construction of an infectious cDNA clone based on a clinically isolated EV71 strain (js1), a virus produced by using the cDNA clone and its derivative clones, and a virus with a reporter gene and The established animal models are used in antiviral drug development, vaccine development, and virus diagnosis.

背景技术Background technique

现有技术公开了肠道病毒是一类病毒的总称,包括脊髓灰质炎病毒(Poliovirus)的3个分型、柯萨奇病毒(Coxsackie virus A)的23个分型、柯萨奇(Coxsackie virus B)病毒的6个分型、埃可病毒(ECHO virus)的31个分型、肠病毒(Enterovirus)的68-71个分型,共67种类型。传统分型以后发现的肠道病毒﹐按照发现顺序命名,现在已经发现的新型肠道病毒有68﹑69﹑70﹑71和72型肠道病毒。新型肠道病毒71型,简称EV71,属于小RNA病毒科,肠道病毒属,其于1969年从澳大利亚和美国分离出,于1973在日本分离出,并被认为是儿童手足口病爆发流行的主要病原体(Schmidt et al.J Infect Dis 1974,129:304-309;Hagiwara et al.Intervirology 1978,9:60-63.)。在1988年以前,EV71病毒主要在美国、日本、欧洲和澳大利亚地区引起婴幼儿手足口病的爆发流行(Weng et al.MicrobesInfect 2010;12:505-10;Tagaya et al.Jpn J Med Sci Biol 1975;28:231-4;Blomberget al.Lancet 1974;2:112;Nagy et al..Arch Virol 1982;71:217-27;Kennett etal.Bull World Health Organ 1974;51:609-15;Gilbert et al.Pediatr Infect Dis J1988;7:484-8)。自1990年以来,EV71病毒在亚洲-太平洋地区引起一系列的爆发流行(Chanet al.Clin Infect Dis 2000;31:678-83;Tu et al.Emerg Infect Dis 2007;13:1733-41;Jeong et al..Arch Virol 2010;155:1707-12)。截止2014年,EV71感染已波及到全球各个洲和国家。研究显示,EV71感染主要分布在亚洲-太平洋地区,在北美、南美、欧洲和澳大利亚也有EV71感染的分布。The prior art discloses that enterovirus is a general term for a class of viruses, including 3 types of poliovirus (Poliovirus), 23 types of Coxsackie virus (Coxsackie virus A), Coxsackie virus (Coxsackie virus A) B) 6 types of viruses, 31 types of ECHO virus, 68-71 types of enterovirus, a total of 67 types. Enteroviruses discovered after traditional typing are named according to the order of discovery. The new types of enteroviruses that have been discovered now include enteroviruses 68, 69, 70, 71 and 72. Novel enterovirus type 71, or EV71 for short, belongs to the Picornaviridae family, the genus Enterovirus, which was isolated from Australia and the United States in 1969, and was isolated in Japan in 1973, and is considered to be the epicenter of the outbreak of hand, foot and mouth disease in children. Major pathogen (Schmidt et al. J Infect Dis 1974, 129:304-309; Hagiwara et al. Intervirology 1978, 9:60-63.). Before 1988, EV71 virus mainly caused outbreaks of hand, foot and mouth disease in infants and young children in the United States, Japan, Europe and Australia (Weng et al. Microbes Infect 2010; 12: 505-10; Tagaya et al. Jpn J Med Sci Biol 1975 ;28:231-4;Blomberg et al.Lancet 1974;2:112;Nagy et al..Arch Virol 1982;71:217-27;Kennett et al.Bull World Health Organ 1974;51:609-15;Gilbert et al . Pediatr Infect Dis J 1988;7:484-8). Since 1990, EV71 virus has caused a series of outbreaks in the Asia-Pacific region (Chanet al. Clin Infect Dis 2000; 31: 678-83; Tu et al. Emerg Infect Dis 2007; 13: 1733-41; Jeong et al. al.. Arch Virol 2010;155:1707-12). As of 2014, EV71 infection has spread to all continents and countries around the world. Studies have shown that EV71 infection is mainly distributed in the Asia-Pacific region, and EV71 infection is also distributed in North America, South America, Europe and Australia.

研究报道了EV71病毒是单股正链RNA病毒,其基因组可以编码一条单长的开放阅读框(ORF),在基因组的两段还包含两个长的非编码区5'TURs和3'TURs。5'TURs含有核糖体内部结合位点(Internal ribosome entry site,IRES),启动病毒的翻译过程(Hellen etal.Genes Dev.2001;15,1593–1612)。病毒编码的开放阅读框翻译后被病毒自身编码的蛋白酶切割加工成单个病毒蛋白,其中包括组成病毒颗粒的结构蛋白VP4,VP2,VP3,VP1和负责病毒复制的非结构蛋白2A,2B,2C,3A,3B,3C及3D(Racaniello,et al.FieldsVirology.2007,Fifth edition)。The study reported that EV71 virus is a single-stranded positive-stranded RNA virus whose genome can encode a single-length open reading frame (ORF), and also contains two long non-coding regions 5'TURs and 3'TURs in two segments of the genome. 5'TURs contain internal ribosome entry sites (IRES) that initiate viral translation (Hellen et al. Genes Dev. 2001; 15, 1593–1612). After translation, the virus-encoded open reading frame is cleaved and processed into a single viral protein by protease encoded by the virus itself, including the structural proteins VP4, VP2, VP3, VP1 that make up the virus particle and the non-structural proteins 2A, 2B, 2C responsible for viral replication, 3A, 3B, 3C and 3D (Racaniello, et al. FieldsVirology. 2007, Fifth edition).

研究报道,灵长类动物可以作为EV71的感染模型。最早在1978年,Hashimoto等报道使用1.8-3.8kg的食蟹猴,在隔离了9周以后,可以感染来自一名3岁儿童粪便标本分离到的EV71病毒毒株,EV71病毒对这种猴子具有神经毒性,在感染的第四天食蟹猴表现出神经损伤的临床症状,损伤程度跟病毒滴度呈正相关。并且EV71病毒可以诱发猴子产生血清中和抗体(Hashimoto et al.Arch Virol.1978;56:257-61)。Zhang等使用3-3.5岁大小的恒河猴可以建立出现脑内感染,肺水肿、出血伴发神经损伤等症状的动物感染模型,而静脉和呼吸系统感染可直接导致神经系统感染。因此,通过不同的感染途径可以获得不同研究目的的模型(Zhang et al.Lab Invest.2011;91:1337-50)。此外还有能引起中枢神经系统疾病的恒河猴动物模型,(Liu et al.Virology.2011;412:91-100)。Studies have reported that primates can be used as an infection model for EV71. As early as 1978, Hashimoto et al. reported that 1.8-3.8kg cynomolgus monkeys could be infected with the EV71 virus strain isolated from the fecal specimen of a 3-year-old child after 9 weeks of isolation. Neurotoxicity, cynomolgus monkeys showed clinical symptoms of nerve damage on the fourth day of infection, and the degree of damage was positively correlated with virus titer. And EV71 virus can induce monkeys to produce serum neutralizing antibodies (Hashimoto et al. Arch Virol. 1978; 56: 257-61). Zhang et al. used 3-3.5-year-old rhesus monkeys to establish an animal infection model with symptoms such as brain infection, pulmonary edema, and hemorrhage accompanied by nerve damage. Intravenous and respiratory system infections can directly lead to nervous system infections. Therefore, models for different research purposes can be obtained through different infection routes (Zhang et al. Lab Invest. 2011;91:1337-50). In addition, there are rhesus monkey animal models that can cause central nervous system diseases, (Liu et al. Virology. 2011; 412:91-100).

EV71的非灵长类动物模型也有报道,如,老鼠适应性突变的EV71毒株EV71/MP4可以感染ICR老鼠,出现神经和肺部损伤(Chen et al.J Virol.2007,81:8996-9003;Wang etal.J Virol.2004,78:7916-24)。Arita等使用免疫缺陷的非肥胖严重糖尿病的小鼠(NOD/SCID mice),对病毒传代获得了可以使3周大小NOD/SCID小鼠感染的鼠适应EV71毒株,该小鼠模型自然杀伤细胞功能被抑制,并且缺乏功能性T、B细胞。而且获得的鼠适应株主要感染动物的中枢神经系统、心脏和骨骼肌(Arita et al.J Virol.2008,82(4):1787-97)。利用干扰素受体α、β和γ缺陷免疫缺陷小鼠AG129小鼠,2周龄或更小龄的AG129小鼠可以感染EV71天然毒株,并且在小鼠死亡之前表现出肢体瘫痪的症状(Khong et al.J Virol.2012,86(4):2121-31)。三周龄的表达有EV71受体hSCARB2的转基因小鼠可以成功感染EV71Isehara/Japan/99(Isehara)毒株;研究表明构建EV71的老鼠模型需要特殊的老鼠适应性毒株或基因缺失或修饰的老鼠。Non-primate models of EV71 have also been reported, for example, the mouse adaptive mutant EV71 strain EV71/MP4 can infect ICR mice with neurological and lung damage (Chen et al. J Virol. 2007, 81:8996-9003 ; Wang et al. J Virol. 2004, 78:7916-24). Arita et al. used immunodeficient non-obese severely diabetic mice (NOD/SCID mice) to passage the virus to obtain a mouse-adapted EV71 strain that can infect 3-week-old NOD/SCID mice. This mouse model is a natural killer cell Function is suppressed and functional T and B cells are absent. Moreover, the obtained murine-adapted strain mainly infects the central nervous system, heart and skeletal muscle of animals (Arita et al. J Virol. 2008, 82(4): 1787-97). Using AG129 mice, immunodeficient mice deficient in interferon receptor alpha, beta and gamma, AG129 mice 2 weeks of age or younger can be infected with the natural strain of EV71 and show symptoms of paralysis before the mice die ( Khong et al. J Virol. 2012, 86(4):2121-31). Three-week-old transgenic mice expressing the EV71 receptor hSCARB2 can be successfully infected with the EV71Isehara/Japan/99 (Isehara) strain; studies have shown that the construction of EV71 mouse models requires special mouse-adapted strains or genetically deleted or modified mice .

研究还报道,单正链(positive-strand)RNA病毒的基因组RNA被释放,进入宿主细胞细胞浆后可以直接作为mRNA模版进行翻译;翻译产生的病毒非结构蛋白招募病毒基因组形成复制复合物起始病毒的基因复制及生活周期,因此单正链RNA病毒的基因组RNA具有感染性,经导入到宿主细胞后,可以完全起始病毒的整个生活周期(Racaniello,etal.Science.1981,214(4523):916)。构建感染性克隆的方法通常采用病毒感染的细胞总RNA作为模版,逆转录成互补DNA(cDNA),然后克隆病毒片段入克隆载体形成病毒的感染性克隆。构建的感染性克隆利用体外转录产生完整的病毒RNA,然后转染病毒RNA入宿主细胞来起始病毒生活周期,产生子代病毒。或者构建的感染性克隆如果带有真核细胞启动子,可以直接转染质粒,由宿主细胞的RNA聚合酶转录出病毒全长RNA,进而起始病毒生活周期,产生子代病毒。The study also reported that the genomic RNA of a single positive-strand RNA virus is released and can be directly translated as an mRNA template after entering the cytoplasm of the host cell; the viral non-structural proteins generated by translation recruit the viral genome to form the initiation of a replication complex. The gene replication and life cycle of the virus, so the genomic RNA of the single positive-stranded RNA virus is infectious, and after being introduced into the host cell, the entire life cycle of the virus can be completely initiated (Racaniello, et al. Science. 1981, 214 (4523) :916). The method for constructing infectious clones usually uses the total RNA of virus-infected cells as a template, reverse-transcribes it into complementary DNA (cDNA), and then clones the viral fragments into a cloning vector to form an infectious clone of the virus. The constructed infectious clones utilize in vitro transcription to generate complete viral RNA, and then transfect the viral RNA into host cells to initiate the viral life cycle and produce progeny viruses. Alternatively, if the constructed infectious clone has a eukaryotic cell promoter, the plasmid can be directly transfected, and the full-length viral RNA is transcribed by the RNA polymerase of the host cell, thereby initiating the viral life cycle and producing progeny viruses.

老鼠模型研究证实EV71的VP1 145位谷氨酸是病毒致小鼠死亡的主要位点,VP2149位赖氨酸的甲基化能够协同促进VP1 145E致小鼠死亡的能力(Huang etal.Virology.2012,422(1):132-43)。此病毒位点在病毒体外传代过程中易突变未145G,导致病毒感染动物能力的下降(Yi et al.Unpublished data)。Mouse model studies have confirmed that glutamate at position 145 of EV71 is the main site of virus-induced mouse death, and methylation of lysine at position 149 of VP2 can synergistically promote the ability of VP1 145E to cause mouse death (Huang et al.Virology.2012 , 422(1):132-43). This viral site is easily mutated to 145G during the in vitro passage of the virus, resulting in a decrease in the ability of the virus to infect animals (Yi et al. Unpublished data).

基于现有技术的基础与现状,本申请的发明人拟提供基于EV71毒株的感染性cDNA克隆及其应用。Based on the basis and status of the prior art, the inventors of the present application intend to provide an infectious cDNA clone based on EV71 strain and its application.

发明内容SUMMARY OF THE INVENTION

本发明的目的是基于现有技术的基础与现状,提供基于EV71毒株的感染性cDNA克隆及其应用。具体涉及一个稳定的EV71毒株的感染性cDNA克隆,该克隆及其衍生产生的病毒RNA在细胞中能自行复制、产生子代病毒颗粒及表达报道基因。The purpose of the present invention is to provide an infectious cDNA clone based on EV71 strain and its application based on the basis and current state of the prior art. Specifically, it relates to an infectious cDNA clone of a stable EV71 strain, and the clone and its derived viral RNA can self-replicate in cells, produce progeny virus particles and express reporter genes.

本发明要解决的另一个技术问题是提供以上述克隆为基础构建的重组病毒或者亚单位病毒颗粒、质粒等,为构建动物模型、疫苗开发及抗病毒药物的开发提供支持。Another technical problem to be solved by the present invention is to provide recombinant viruses or subunit virus particles, plasmids, etc. constructed on the basis of the above clones, so as to provide support for the construction of animal models, the development of vaccines and the development of antiviral drugs.

本发明从临床上分离到一株EV71毒株(命名为js1),其无需老鼠适应性突变、且能感染未经基因背景改变的小鼠,通过构其感染性克隆能产生稳定基因序列的病毒颗粒,感染普通老鼠,建立简便、高效的EV71动物感染模型。The present invention isolates an EV71 strain (named js1) from the clinic, which does not require mouse adaptive mutation and can infect mice without genetic background changes, and can produce a virus with stable gene sequence by constructing its infectious clone particles, infect ordinary mice, and establish a simple and efficient EV71 animal infection model.

更具体的,more specific,

本发明提供了一种cDNA,它包含EV71毒株的核酸序列和一个低拷贝质粒骨架的核酸序列;其中,EV71毒株的核酸序列涵盖EV71病毒5′到3′正向极性序列,包含病毒5′及3′非编码区及一个编码病毒蛋白的开放阅读框。The present invention provides a cDNA comprising the nucleic acid sequence of the EV71 strain and the nucleic acid sequence of a low-copy plasmid backbone; wherein, the nucleic acid sequence of the EV71 strain covers the 5' to 3' forward polar sequence of the EV71 virus, including the virus 5' and 3' non-coding regions and an open reading frame encoding a viral protein.

较好的,它还包括EV71毒株的核酸序列插入的报道基因荧光素酶或者荧光蛋白的序列。Preferably, it also includes the sequence of reporter gene luciferase or fluorescent protein inserted into the nucleic acid sequence of EV71 strain.

所述的病毒蛋白开放阅读框的氨基酸序列如SEQ ID NO 4所示。The amino acid sequence of the viral protein open reading frame is shown in SEQ ID NO 4.

所述的低拷贝质粒骨架的编码序列如SEQ ID NO 3所示。The coding sequence of the low-copy plasmid backbone is shown in SEQ ID NO 3.

所述的EV71毒株的核酸序列如SEQ ID NO 2所示。The nucleic acid sequence of the EV71 strain is shown in SEQ ID NO 2.

在本发明的一个优选实施例中,所述的EV71毒株的感染性cDNA克隆,其序列如SEQID NO 1所示。In a preferred embodiment of the present invention, the infectious cDNA clone of the EV71 strain has the sequence shown in SEQ ID NO 1.

在本发明的一个实施例中,提供了一个稳定的、一株临床分离的EV71毒株的感染性cDNA克隆的构建(核酸序列1)及其含有各类报道基因的衍生克隆(核酸序列5,核酸序列6)、及以其为母本构建的各种突变克隆。这些克隆产生的病毒RNA在细胞中能自行复制、产生子代病毒颗粒及表达报道基因。In one embodiment of the present invention, the construction of a stable infectious cDNA clone of a clinically isolated EV71 strain (nucleic acid sequence 1) and its derivative clones containing various reporter genes (nucleic acid sequence 5, Nucleic acid sequence 6), and various mutant clones constructed therefrom. The viral RNA produced by these clones can self-replicate in cells, produce progeny virus particles and express reporter genes.

本发明还包括以核酸序列6或核酸序列7所述的序列为母本,通过替代Nluc或EGFP所构建的含有异源性报道序列或目的基因的重组病毒克隆及其序列。The present invention also includes recombinant virus clones containing heterologous reporter sequences or target genes constructed by replacing Nluc or EGFP with the sequences described in nucleic acid sequence 6 or nucleic acid sequence 7 as the parent and their sequences.

本发明还包括各种嵌合病毒感染性克隆和含有异源性报道序列或目的基因的重组病毒克隆产生的各种嵌合病毒和含有报道基因或外源基因的各种病毒颗粒。The present invention also includes various chimeric virus infectious clones and various chimeric viruses produced by recombinant virus clones containing heterologous reporter sequences or genes of interest, and various viral particles containing reporter genes or foreign genes.

本发明还包括全长感染性克隆序列构建的在病毒蛋白中同开放阅读框插入有异源性抗性序列的重组病毒克隆及其序列。The present invention also includes recombinant virus clones constructed from full-length infectious clone sequences and inserted into the viral protein with a heterologous resistance sequence in the same open reading frame and their sequences.

具体的,本发明提供了一种包含临床分离的EV71毒株(js1)的感染性cDNA克隆(核酸序列1),此感染性克隆(核酸序列1)包含一个全长的EV71毒株(js1)的核酸序列(核酸序列2)及一个低拷贝质粒骨架(核酸序列3)。核酸序列2涵盖EV71病毒5′到3′正向极性(positive-sense)序列,其中包含病毒5′及3′非编码区及一个编码病毒蛋白的开放阅读框(open reading frame),开放阅读框病毒编码蛋白(蛋白序列4),在此感染性克隆(核酸序列1)中插入报道基因荧光素酶NanoLuc(Nluc)及荧光蛋白EGFP,分别构成带有Nluc的感染性克隆(核酸序列5)及带有EGFP的感染性克隆(核酸序列6),以及以这些克隆为基础,通过改变核酸的手段得到的突变病毒克隆(adapted virus),减毒病毒克隆(live-attenuatedvirus),复制缺陷病毒克隆(defective virus)及复制性的非感染性克隆(replication-competent non-infectious virus)等衍生物(derivative),如包括缺失结构蛋白的亚基因组复制子。Specifically, the present invention provides an infectious cDNA clone (nucleic acid sequence 1) comprising a clinically isolated EV71 strain (js1), the infectious clone (nucleic acid sequence 1) comprising a full-length EV71 strain (js1) nucleotide sequence (nucleotide sequence 2) and a low-copy plasmid backbone (nucleotide sequence 3). Nucleic acid sequence 2 covers the 5' to 3' positive-sense sequence of the EV71 virus, which includes the 5' and 3' non-coding regions of the virus and an open reading frame (open reading frame) encoding a viral protein, open reading Frame virus encoding protein (protein sequence 4), insert reporter gene luciferase NanoLuc (Nluc) and fluorescent protein EGFP into this infectious clone (nucleic acid sequence 1), respectively constitute an infectious clone with Nluc (nucleic acid sequence 5) And infectious clones with EGFP (nucleic acid sequence 6), and mutant virus clones (adapted virus), attenuated virus clones (live-attenuated virus), replication-defective virus clones obtained by means of changing nucleic acid based on these clones Derivatives such as defective virus and replication-competent non-infectious virus, such as subgenomic replicons including deletion of structural proteins.

上述序列1-6具体如下:The above sequences 1-6 are as follows:

核酸序列1,SEQ ID NO 1:Nucleic acid sequence 1, SEQ ID NO 1:

GCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCTAATACGACTCACTATAGGTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACATGGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAaagcttatGCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCAC CTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCTAATACGACTCACTAT AGGTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACATGGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTA ACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCAT GGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCC GGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATA GGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTT CAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAAC CCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACC ATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAaagcttat

核酸序列2,SEQ ID NO 2:Nucleic acid sequence 2, SEQ ID NO 2:

TTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACATGGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACATGGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACT ATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGC AATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGC GCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGT TGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAG AACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCG CACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATG ACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGC

核酸序列3,SEQ ID NO 3:Nucleic acid sequence 3, SEQ ID NO 3:

AGCGCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCAGCGCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGG CACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGC

蛋白序列4,SEQ ID NO 4:Protein sequence 4, SEQ ID NO 4:

MGSQVSTQRSGSYENSNSATEGSTINYTTINYYKDSYAATAGKQSLKQDPDKFANPVKDIFTEMAAPLKSPSAEACGYSDRVAQLTIGNSTITTQEAANIIVGYGEWPSYCSDSDATAVDKPTRPDVSVNRFYTLDTKLWEKSSKGWYWKFPDVLTETGVFGQNAQFHYLYRSGFCIHVQCNASKFHQGALLVAVLPEYVIGTVAGGTGTEDTHPPYKQTQPGADGFELQHPYVLDAGIPISQLTVCPHQWINLRTNNCATIIVPYINALPFDSALNHCNFGLLVVPISPLDYDQGATPVIPITITLAPMCSEFAGLRQAVTQGFPTELKPGTNQFLTTDDGVSAPILPNFHPTPCIHIPGEVRNLLELCQVETILEVNNVPTNATSLMERLRFPVSAQAGKGELCAVFRADPGRNGPWQSTLLGQLCGYYTQWSGSLEVTFMFTGSFMATGKMLIAYTPPGGPLPKDRATAMLGTHVIWDFGLQSSVTLVIPWISNTHYRAHARDGVFDYYTTGLVSIWYQTNYVVPIGAPNTAYIIALAAAQKNFTMKLCKDASDILQTGTIQGDRVADVIESSIGDSVSRALTHALPAPTGQNTQVSSHRLDTGKVPALQAAEIGASSNASDESMIETRCVLNSHSTAETTLDSFFSRAGLVGEIDLPLEGTTNPNGYANWDIDITGYAQMRRKVELFTYMRFDAEFTFVACTPTGEVVPQLLQYMFVPPGAPKPDSRESLAWQTATNPSVFVKLSDPPAQVSVPFMSPASAYQWFYDGYPTFGEHKQEKDLEYGACPNNMMGTFSVRTVGTSKSKYPLVVRIYMRMKHVRAWIPRPMRNQNYLFKANPNYAGNSIKPTGASRTAITTLGKFGQQSGAIYVGNFRVVNRHLATHNDWANLVWEDSSRDLLVSSTTAQGCDTIARCDCQTGVYYCNSMRKHYPVSFSKPSLIYVEASEYYPARYQSHLMLAQGHSEPGDCGGILRCQHGVIGIVSTGGNGLVGFADVRDLLWLDEEAMEQGVSDYIKGLGDAFGTGFTDAVSREVEALKNYLIGSEGAVEKILKNLIKLISALVIVIRSDYDMVTLTATLALIGCHGSPWAWIKAKTASILGIPIAQKQSASWLKKFNDMANAAKGLEWVSNKISKFIDWLKEKIVPAAREKVEFLNNLKQLPLLENQISNLEQSAASQEDLEVMFGNVSYLAHFCRKFQPLYATEAKRVYALEKRMNNYMQFKSKHRIEPVCLIIRGSPGTGKSLATGIIARAIADKYHSSVYSLPPDPDHFDGYKQQVVTVMDDLCQNPDGKDMSLFCQMVSTVDFIPPMASLEEKGVSFTSKFVIASTNASNIIVPTVSDSDAIRRRFYMDCDIEVTDSYKTDLGRLDAGRAAKLCSENNTANFKRCSPLVCGKAIQLRDRKSKVRYSVDTVVSELIREYSNRSAIGNTIEALFQGPPKFRPIRISLEEKPAPDAISDLLASVDSEEVRQYCRDQGWIIPEAPTNVERHLNRAVLVMQSITTVVAVVSLVYVIYKLFAGFQGAYSGAPKQVLKKPALRTATVQGPSLDFALSLLRRNIRQVQTDQGHFTMLGVRDRLAVLPRHSQPGKTIWIEHKLVNVLDAVELVDEQGVNLELTLITLDTNEKFRDITKFIPENISTASDATLVINTEHMPSMFVPVGDVVQYGFLNLSGKPTHRTMMYNFPTKAGQCGGVVTSVGKVVGIHIGGNGRQGFCAGLKRSYFASEQGEIQWVKPNKETGRLNINGPTRTKLEPSVFHDIFEGNKEPAVLHSKDPRLEVDFEQALFSKYVGNTLHEPDEYIKEAALHYANQLKQLEINTSQMSMEEACYGTENLEAIDLHTSAGYPYSALGIKKRDILDPTTRDVSRMKFYMDKYGLDLPYSTYVKDELRSIDKIKKGKSRLIEASSLNDSVYLRMAFGHLYEAFHANPGTITGSAVGCNPDTFWSKLPILLPGSLFAFDYSGYDASLSPVWFRALELVLREIGYSEEAISLIEGINHTHHVYRNKTYCVLGGMPSGCSGTSIFNSMINNIIIRALLIKTFKGIDLDELNMVAYGDDVLASYPFPIDCLELAKTGKEYGLTMTPADKSPCFNEVNWGNATFLKRGFLPDEQFPFLIHPTMPMREIHESIRWTKDARNTQDHVRSLCLLAWHNGKQEYEKFVSTIRSVPVGRALAIPNYENLRRNWLELFMGSQVSTQRSGSYENSNSATEGSTINYTTINYYKDSYAATAGKQSLKQDPDKFANPVKDIFTEMAAPLKSPSAEACGYSDRVAQLTIGNSTITTQEAANIIVGYGEWPSYCSDSDATAVDKPTRPDVSVNRFYTLDTKLWEKSSKGWYWKFPDVLTETGVFGQNAQFHYLYRSGFCIHVQCNASKFHQGALLVAVLPEYVIGTVAGGTGTEDTHPPYKQTQPGADGFELQHPYVLDAGIPISQLTVCPHQWINLRTNNCATIIVPYINALPFDSALNHCNFGLLVVPISPLDYDQGATPVIPITITLAPMCSEFAGLRQAVTQGFPTELKPGTNQFLTTDDGVSAPILPNFHPTPCIHIPGEVRNLLELCQVETILEVNNVPTNATSLMERLRFPVSAQAGKGELCAVFRADPGRNGPWQSTLLGQLCGYYTQWSGSLEVTFMFTGSFMATGKMLIAYTPPGGPLPKDRATAMLGTHVIWDFGLQSSVTLVIPWISNTHYRAHARDGVFDYYTTGLVSIWYQTNYVVPIGAPNTAYIIALAAAQKNFTMKLCKDASDILQTGTIQGDRVADVIESSIGDSVSRALTHALPAPTGQNTQVSSHRLDTGKVPALQAAEIGASSNASDESMIETRCVLNSHSTAETTLDSFFSRAGLVGEIDLPLEGTTNPNGYANWDIDITGYAQMRRKVELFTYMRFDAEFTFVACTPTGEVVPQLLQYMFVPPGAPKPDSRESLAWQTATNPSVFVKLSDPPAQVSVPFMSPASAYQWFYDGYPTFGEHKQEKDLEYGACPNNMMGTFSVRTVGTSKSKYPLVVRIYMRMKHVRAWIPRPMRNQNYLFKANPNYAGNSIKPTGASRTAITTLGKFGQQSGAIYVGNFRVVNRHLATHNDWANLVWEDSSRDLLVSSTTAQGCDTIARCDCQTGVYYCNSMRKHYPVSFSKPSLIYVEASEYYPARYQSHLMLAQGHSEPGDCGGILRCQHGVIGIVSTGGNGLVGFADVR DLLWLDEEAMEQGVSDYIKGLGDAFGTGFTDAVSREVEALKNYLIGSEGAVEKILKNLIKLISALVIVIRSDYDMVTLTATLALIGCHGSPWAWIKAKTASILGIPIAQKQSASWLKKFNDMANAAKGLEWVSNKISKFIDWLKEKIVPAAREKVEFLNNLKQLPLLENQISNLEQSAASQEDLEVMFGNVSYLAHFCRKFQPLYATEAKRVYALEKRMNNYMQFKSKHRIEPVCLIIRGSPGTGKSLATGIIARAIADKYHSSVYSLPPDPDHFDGYKQQVVTVMDDLCQNPDGKDMSLFCQMVSTVDFIPPMASLEEKGVSFTSKFVIASTNASNIIVPTVSDSDAIRRRFYMDCDIEVTDSYKTDLGRLDAGRAAKLCSENNTANFKRCSPLVCGKAIQLRDRKSKVRYSVDTVVSELIREYSNRSAIGNTIEALFQGPPKFRPIRISLEEKPAPDAISDLLASVDSEEVRQYCRDQGWIIPEAPTNVERHLNRAVLVMQSITTVVAVVSLVYVIYKLFAGFQGAYSGAPKQVLKKPALRTATVQGPSLDFALSLLRRNIRQVQTDQGHFTMLGVRDRLAVLPRHSQPGKTIWIEHKLVNVLDAVELVDEQGVNLELTLITLDTNEKFRDITKFIPENISTASDATLVINTEHMPSMFVPVGDVVQYGFLNLSGKPTHRTMMYNFPTKAGQCGGVVTSVGKVVGIHIGGNGRQGFCAGLKRSYFASEQGEIQWVKPNKETGRLNINGPTRTKLEPSVFHDIFEGNKEPAVLHSKDPRLEVDFEQALFSKYVGNTLHEPDEYIKEAALHYANQLKQLEINTSQMSMEEACYGTENLEAIDLHTSAGYPYSALGIKKRDILDPTTRDVSRMKFYMDKYGLDLPYSTYVKDELRSIDKIKKGKSRLIEASSLNDSVYLRMAFGHLYEAFHANPGTITGSAVGCNPDTFWSKLPILLPGSLFAFDYSGYDASLSPVWFRALELVLREIGYSEEAISLIEGI NHTHHVYRNKTYCVLGGMPSGCSGTSIFNSMINNIIIRALLIKTFKGIDLDELNMVAYGDDVLASYPFPIDCLELAKTGKEYGLTMTPADKSPCFNEVNWGNATFLKRGFLPDEQFPFLIHPTMPMREIHESIRWTKDARNTQDHVRSLCLLAWHNGKQEYEKFVSTIRSVPVGRALAIPNYENLRRNWLELF

核酸序列5,SEQ ID NO 5:Nucleic acid sequence 5, SEQ ID NO 5:

GCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCTAATACGACTCACTATAGGTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACtctagaatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgatgcatGCGATCACCACTCTTGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAaagcttatGCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCAC CTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCTAATACGACTCACTAT AGGTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACtctagaatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtac cctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgatgcatGCGATCACCACTCTTGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCC CAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAAT TGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCC ACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGG GCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTT CACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTG ATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAaagcttat

核酸序列6,SEQ ID NO 6:Nucleic acid sequence 6, SEQ ID NO 6:

GCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCTAATACGACTCACTATAGGTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACcatatgATGgtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagatgcatGCGATCACCACTCTTGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACCGATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCGAGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTCAAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCTCGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCATCACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTTGAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCCCAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAaagcttat。GCTAGCGGAGTGTATACTGGCTTACTATGTTGGCACTGATGAGGGTGTCAGTGAAGTGCTTCATGTGGCAGGAGAAAAAAGGCTGCACCGGTGCGTCAGCAGAATATGTGATACAGGATATATTCCGCTTCCTCGCTCACTGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGAAATGGCTTACGAACGGGGCGGAGATTTCCTGGAAGATGCCAGGAAGATACTTAACAGGGAAGTGAGAGGGCCGCGGCAAAGCCGTTTTTCCATAGGCTCCGCCCCCCTGACAAGCATCACGAAATCTGACGCTCAAATCAGTGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCTGGCGGCTCCCTCGTGCGCTCTCCTGTTCCTGCCTTTCGGTTTACCGGTGTCATTCCGCTGTTATGGCCGCGTTTGTCTCATTCCACGCCTGACACTCAGTTCCGGGTAGGCAGTTCGCTCCAAGCTGGACTGTATGCACGAACCCCCCGTTCAGTCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGAAAGACATGCAAAAGCACCACTGGCAGCAGCCACTGGTAATTGATTTAGAGGAGTTAGTCTTGAAGTCATGCGCCGGTTAAGGCTAAACTGAAAGGACAAGTTTTGGTGACTGCGCTCCTCCAAGCCAGTTACCTCGGTTCAAAGAGTTGGTAGCTCAGAGAACCTTCGAAAAACCGCCCTGCAAGGCGGTTTTTTCGTTTTCAGAGCAAGAGATTACGCGCAGACCAAAACGATCTCAAGAAGATCATCTTATTAAGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCAC CTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTGTCGACGCGGCCGCTAATACGACTCACTAT AGGTTAAAACAGCCTGTGGGTTGCACCCACTCACAGGGCCTACTGGGCGCAAGCACTCTGGTACCTCGGTACCTTTGTGCGCCTGTTTTACACCCCCCCCCCAATGAAACTTAGAAGCAATAAACCACGATCAATAGCAGGCATAACGCTCCAGTTATGTCTTGATCAAGCACTTCTGTTTCCCCGGACTGAGTATCAATAGACTGCTCGCGCGGTTGAAGGAGAAAACGTTCGTTATCCGGCTAACTACTTCGGAAAACCTAGTAACACCATGAAAGTTGCGGAGAGCTTCGTTCAGCACTCCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCGTTGGCGGCCTGCCCATGGGGTAACCCATGGGGCGCTCTAATACGGACATGGTGTGAAGAGTCTACTGAGCTAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACTGCGGAGCACACGCCCACAAGCCAGCGGGTAGTGTGTCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATCTTTATATTGGCTGCTTATGGTGACAATTAAAGAATTGTTACCATATAGCTATTGGATTAGCCATCCGGTGTGCAACAGAGCAATTATTTACCTATTTATTGGTTTTGTACCATTAACCTCGAATTCTGTGACCACCCTTAATTATATCTTGACCCTTAACACAGCTAAACcatatgATGgtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgac ttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagatgcatGCGATCACCACTCTTGGTTCGCAAGTGTCTACACAGCGCTCCGGTTCTTACGAAAACTCAAACTCAGCCACTGAGGGTTCTACCATAAACTACACCACCATTAATTACTACAAAGACTCCTATGCTGCCACAGCAGGCAAaCAGAGTCTCAAGCAGGATCCAGACAAGTTTGCAAATCCTGTTAAAGACATATTCACcGAAATGGCAGCGCCACTGAAGTCCCCATCCGCTGAGGCATGTGGATACAGTGATCGAGTGGCGCAATTAACTATTGGCAACTCCACCATCACGACGCAAGAAGCGGCTAACATCATAGTCGGCTATGGTGAGTGGCCTTCCTACTGCTCAGATTCTGACGCTACAGCAGTGGATAAACCAACGCGCCCGGATGTTTCAGTGAACAGGTTTTACACATTGGACACTAAATTGTGGGAGAAATCGTCCAAGGGATGGTACTGGAAGTTCCCGGATGTGTTAACTGAAACTGGGGTTTTTGGGCAAAATGCACAATTCCACTACCTCTACC GATCAGGGTTCTGCATCCACGTGCAGTGCAATGCCAGTAAATTCCACCAAGGAgCACTcCtAgTCGCTGTCCTACCAGAGTATGTCATTGGGACAGTGGCAGGCGGTACAGGGACGGAAGACACCCACCCCCCCTACAAGCAGACCCAACCCGGCGCCGATGGTTTCGAGTTGCAACACCCGTACGTGCTTGATGCTGGCATCCCAATATCACAGTTAACAGTGTGCCCACACCAGTGGATTAATTTGAGGACCAACAATTGTGCTACAATAATAGTGCCATACATTAACGCACTGCCTTTTGATTCTGCCTTGAACCATTGCAACTTTGGCCTGTTAGTTGTGCCTATTAGCCCACTAGACTACGACCAAGGAGCAACGCCAGTAATCCCTATAACTATCACATTGGCCCCAATGTGCTCTGAATTCGCAGGTCTTAGGCAGGCAGTCACGCAAGGGTTCCCCACCGAGCTAAAACCTGGCACAAATCAATTTTTAACCACCGATGATGGCGTCTCAGCACCTATTCTACCAAACTTCCACCCCACCCCGTGTATCCACATACCTGGTGAAGTTAGGAACTTGCTAGAGTTATGCCAGGTGGAGACCATTCTGGAGGTTAACAATGTGCCCACGAATGCCACTAGCTTAATGGAGAGACTGCGCTTCCCGGTCTCAGCACAAGCAGGGAAAGGTGAACTGTGTGCGGTGTTTAGAGCCGATCCTGGGCGAAATGGACCATGGCAATCCACCTTACTGGGCCAGTTGTGCGGGTACTACACCCAATGGTCAGGGTCATTGGAAGTCACCTTCATGTTTACTGGATCCTTCATGGCTACCGGCAAGATGCTCATAGCCTATACACCGCCAGGGGGTCCTCTGCCCAAGGACCGGGCGACCGCCATGTTGGGCACGCACGTCATCTGGGATTTTGGGCTGCAATCGTCTGTTACCCTTGTAATACCATGGATCAGTAACACTCATTATAGAGCACATGCCCG AGATGGAGTGTTTGACTATTACACTACAGGGTTAGTCAGTATATGGTACCAGACAAATTACGTGGTTCCAATCGGTGCGCCCAACACAGCCTATATAATAGCACTAGCGGCAGCCCAAAAGAACTTCACTATGAAATTGTGCAAGGATGCTAGTGATATCCTGCAGACGGGCACCATCCAGGGAGATAGGGTGGCAGATGTAATTGAAAGTTCCATAGGAGATAGCGTGAGCAGAGCCCTCACTCACGCTCTACCAGCACCCACAGGCCAAAACACACAGGTGAGCAGTCATCGACTGGATACAGGCAAGGTTCCAGCACTCCAAGCTGCTGAAATTGGGGCATCATCAAATGCTAGTGACGAGAGCATGATTGAAACACGTTGTGTTCTTAACTCGCATAGTACAGCTGAGACCACTCTTGATAGTTTCTTCAGTAGGGCAGGATTAGTTGGAGAGATAGATCTCCCTCTTGAGGGCACAACTAACCCAAATGGTTATGCCAACTGGGACATAGATATAACAGGTTACGCGCAAATGCGTAGAAAGGTAGAGCTATTCACCTACATGCGTTTTGATGCAGAGTTCACTTTTGTTGCGTGCACACCCACCGGGGAGGTTGTCCCACAATTGCTCCAATATATGTTTGTGCCACCTGGAGCCCCTAAGCCAGATTCTAGGGAATCCCTTGCATGGCAAACCGCCACCAACCCCTCAGTTTTTGTCAAGCTGTCAGACCCTCCGGCGCAGGTTTCAGTGCCATTCATGTCACCTGCGAGTGCTTATCAATGGTTTTATGACGGATATCCCACATTCGGAGAACACAAACAGGAGAAAGACCTTGAATACGGGGCATGTCCTAATAACATGATGGGTACATTCTCAGTGCGGACTGTGGGGACCTCCAAGTCCAAGTACCCTTTAGTGGTTAGGATTTACATGAGAATGAAGCACGTCAGGGCGTGGATACCTCGCCCGATGCGCAACCAGAACTACCTGTTC AAAGCCAACCCAAATTATGCTGGCAACTCTATTAAGCCAACTGGTGCCAGTCGCACAGCGATCACCACTCTTGGGAAATTTGGACAACAGTCTGGGGCTATTTATGTGGGCAACTTTAGAGTGGTCAACCGACATCTTGCCACCCATAATGATTGGGCAAATCTTGTTTGGGAAGACAGCTCTCGCGACTTGCTCGTGTCATCCACCACTGCCCAAGGTTGTGACACGATTGCCCGTTGCGATTGCCAGACAGGGGTGTACTACTGTAACTCGATGAGAAAACACTACCCAGTCAGTTTTTCAAAACCCAGCCTGATCTATGTAGAGGCTAGCGAGTATTACCCAGCCAGGTACCAATCACATCTCATGCTCGCACAGGGTCACTCGGAACCTGGTGATTGCGGTGGTATCCTTAGGTGCCAACATGGCGTCATCGGCATAGTGTCTACTGGTGGCAATGGGCTCGTTGGCTTTGCAGACGTCAGAGACCTCTTGTGGTTAGATGAAGAAGCTATGGAACAGGGCGTGTCCGACTACATTAAGGGTCTCGGAGATGCTTTTGGAACAGGCTTCACTGACGCAGTCTCAAGGGAGGTTGAAGCTCTCAAGAACTATCTTATAGGGTCTGAAGGAGCAGTTGAGAAAATTTTGAAAAATCTTATTAAACTAATCTCTGCACTGGTGATTGTGATCAGAAGTGATTACGACATGGTTACCCTCACTGCAACCTTAGCGCTGATAGGTTGTCATGGCAGTCCTTGGGCTTGGATTAAAGCCAAAACAGCCTCCATCTTAGGTATCCCTATCGCCCAAAAGCAGAGCGCTTCCTGGCTCAAGAAGTTCAATGACATGGCCAACGCCGCTAAGGGGTTAGAGTGGGTTTCCAACAAGATCAGCAAATTTATTGATTGGCTTAAGGAGAAAATAGTACCAGCAGCCAGGGAGAAGGTTGAATTCCTAAATAACTTGAAACAGCTGCCACTGCTAGAGAATCAGATCT CGAACTTGGAACAATCTGCTGCTTCACAAGAGGACCTTGAAGTCATGTTTGGGAATGTGTCGTACCTAGCTCACTTCTGTCGCAAGTTTCAACCGCTATACGCCACGGAAGCTAAAAGAGTCTATGCCCTGGAGAAGAGAATGAATAACTATATGCAGTTCAAGAGCAAACACCGAATTGAACCTGTATGTCTCATTATTAGGGGCTCACCAGGCACCGGGAAGTCTCTAGCCACTGGTATTATTGCTCGAGCAATCGCTGATAAGTACCACTCCAGCGTGTACTCGCTCCCACCAGACCCGGATCATTTTGACGGTTACAAGCAACAGGTGGTTACAGTGATGGATGATTTGTGTCAAAACCCCGATGGTAAGGATATGTCCTTATTCTGTCAAATGGTATCCACCGTAGATTTCATTCCACCAATGGCTTCTCTCGAGGAGAAGGGAGTTTCCTTCACCTCTAAGTTTGTCATCGCATCCACTAATGCCAGTAATATCATAGTACCAACAGTGTCTGATTCTGACGCTATTCGCCGCAGGTTCTACATGGACTGTGACATTGAAGTGACAGACTCGTACAAAACAGATCTAGGTAGACTGGATGCAGGGCGAGCCGCTAAACTGTGTTCTGAAAATAACACTGCAAATTTCAAACGTTGCAGCCCATTAGTGTGTGGGAAAGCCATCCAACTTAGAGATAGAAAGTCTAAAGTCAGATACAGTGTGGATACGGTGGTTTCAGAACTTATTAGGGAATACAGCAATAGGTCCGCCATTGGTAACACAATCGAGGCTCTTTTCCAAGGTCCACCCAAGTTCAGGCCAATTAGGATTAGCCTTGAAGAAAAACCAGCCCCAGACGCTATTAGCGATCTCCTTGCTAGTGTAGATAGTGAAGAAGTGCGCCAGTACTGCAGGGATCAAGGCTGGATTATTCCTGAAGCTCCCACCAATGTGGAGCGGCACCTTAATAGAGCGGTGCTCGTCATGCAATCCAT CACCACAGTAGTGGCGGTTGTTTCGTTGGTGTACGTCATCTACAAGCTCTTTGCAGGGTTTCAGGGTGCATATTCTGGTGCTCCTAAGCAAGTGCTTAAGAAACCTGCTCTTCGCACAGCAACAGTGCAGGGTCCGAGCCTTGACTTTGCTCTCTCCCTACTGAGAAGGAACATCAGGCAGGTCCAAACAGACCAAGGGCATTTCACCATGTTGGGTGTTAGGGATCGCTTAGCAGTCCTCCCACGCCACTCACAACCTGGCAAAACCATTTGGATTGAGCACAAACTCGTGAACGTCCTTGATGCAGTTGAACTGGTGGATGAGCAAGGAGTCAACCTGGAATTAACCCTCATCACTCTTGACACCAACGAGAAGTTTAGGGATATCACCAAATTCATCCCAGAAAATATCAGCACTGCTAGCGATGCCACCCTAGTGATCAACACGGAGCACATGCCGTCAATGTTTGTCCCGGTGGGTGACGTTGTGCAGTATGGCTTTTTGAATCTCAGTGGCAAGCCTACCCATCGCACCATGATGTACAATTTTCCTACTAAAGCAGGACAGTGTGGAGGAGTGGTGACATCTGTTGGGAAGGTTGTCGGTATTCACATTGGTGGCAATGGCAGACAAGGTTTTTGCGCAGGCCTCAAAAGGAGTTACTTTGCTAGTGAACAAGGAGAGATCCAGTGGGTTAAGCCCAATAAAGAAAcTggAAGACTCAACATCAATGGACCAACCCGCACCAAGTTAGAACCTAGTGTATTCCATGACATCTTCGAGGGAAATAAGGAACCAGCTGTCTTGCACAGTAAAGACCCCCGACTTGAGGTAGATTTTGAACAGGCCCTGTTCTCTAAGTATGTGGGAAACACACTACATGAGCCTGACGAGTACATCAAAGAGGCAGCTCTACATTATGCAAACCAATTAAAGCAACTAGAAATCAATACCTCTCAAATGAGCATGGAGGAGGCCTGCTATGGTACTGAGAATCTT GAGGCTATTGATCTTCACACTAGTGCAGGTTACCCCTATAGTGCCCTAGGGATAAAGAAAAGAGACATCTTAGACCCTACCACCAGGGACGTGAGTAGAATGAAGTTCTACATGGACAAGTATGGTCTTGATCTTCCCTACTCCACTTATGTCAAGGACGAGCTACGCTCGATTGATAAAATCAAGAAAGGGAAGTCCCGCCTGATCGAGGCCAGTAGTCTAAATGATTCAGTGTACCTCAGAATGGCTTTCGGGCATTTGTATGAGGCTTTCCACGCAAATCCTGGGACGATAACTGGATCGGCCGTGGGGTGTAACCCTGACACATTCTGGAGCAAGCTGCCAATTTTGCTCCCTGGTTCACTCTTTGCCTTTGACTACTCAGGCTATGATGCCAGCCTTAGCCCTGTCTGGTTCAGAGCATTAGAATTGGTTCTTAGGGAGATAGGGTATAGTGAAGAGGCAATCTCACTCATTGAGGGAATCAACCACACACATCATGTGTATCGTAATAAGACCTATTGCGTGCTTGGTGGGATGCCCTCAGGCTGTTCAGGAACATCCATCTTCAACTCAATGATCAACAACATTATTATCAGAGCACTGCTCATAAAAACATTTAAGGGCATTGATTTGGATGAACTCAACATGGTCGCTTATGGAGACGATGTGCTCGCTAGCTATCCCTTCCCAATTGATTGCTTGGAACTAGCAAAGACTGGTAAGGAGTATGGTCTGACCATGACCCCTGCTGATAAATCTCCTTGCTTTAATGAGGTCAATTGGGGTAATGCGACCTTCCTCAAAAGGGGCTTTTTGCCCGATGAACAGTTTCCATTTTTGATTCACCCTACTATGCCAATGAGGGAGATCCATGAGTCCATTCGATGGACCAAGGACGCACGGAACACTCAAGATCATGTGCGGTCCTTGTGCCTCCTAGCATGGCATAATGGTAAGCAAGAATACGAGAAGTTTGTGAGCACAATTAGGTCTGTCC CAGTAGGGAGAGCGTTGGCTATTCCAAATTATGAAAATCTTAGACGAAATTGGCTCGAGTTATTTTAGAGGTTATACACACCTCAACCCCACCAGAAATCTGGTCGTGAATGTGACTGGTGGGGGTAAATTTGTTATAACCAGAATAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAaagcttat.

本发明还包括这些cDNA克隆的表达产物。The present invention also includes the expression products of these cDNA clones.

本发明还包括含有上述cDNA的双链DNA,能产生全长感染性克隆序列的双链DNA(double stranded DNA),正向cDNA(positive-sense cDNA)或负向cDNA(negative-sensecDNA)。The present invention also includes double-stranded DNA containing the above cDNA, double stranded DNA capable of producing a full-length infectious clone sequence, positive-sense cDNA or negative-sense cDNA.

本发明还包括含有上述cDNA或者双链DNA的质粒。The present invention also includes plasmids containing the above-mentioned cDNA or double-stranded DNA.

较好的,该质粒能够转录产生EV71毒株的全长感染性RNA或者其突变体,能通过体外转录产生含有全长EV71毒株(js1)全长感染性RNA的质粒,能通过体外转录产生含有全长EV71毒株(js1)全长感染性RNA的质粒及衍生质粒。Preferably, the plasmid can be transcribed to produce the full-length infectious RNA of the EV71 strain or its mutant, can be transcribed in vitro to produce a plasmid containing the full-length infectious RNA of the full-length EV71 strain (js1), and can be produced by in vitro transcription. Plasmids and derivatives containing the full-length infectious RNA of the EV71 strain (js1).

其中,衍生质粒包括:Among them, the derivative plasmids include:

A.利用其他分离株(isolates)的部分序列替换权利要求1中EV71毒株(js1)全长感染克隆的部分序列得到的重组病毒克隆;A. The recombinant virus clone obtained by replacing the partial sequence of the full-length infection clone of EV71 strain (js1) in claim 1 with the partial sequence of other isolates;

B.利用基因突变对权利要求1或2中EV71毒株(js1)全长感染克隆中的序列进行突变得到的突变病毒克隆;B. a mutant virus clone obtained by mutating the sequence in the full-length infection clone of EV71 strain (js1) in claim 1 or 2 by gene mutation;

C.由EV71毒株(js1)全长感染克隆产生的病毒经过适应性突变产生的减毒(live-attenuated),复制非感染病毒(replication competent non-infectious)及非复制性病毒(defective variants)等衍生克隆。C. Live-attenuated, replication competent non-infectious and defective variants of virus produced by full-length infected clones of EV71 strain (js1) and other derivative clones.

本发明提供了一种质粒,它含有上述的双链DNA或者其衍生物。The present invention provides a plasmid containing the above-mentioned double-stranded DNA or its derivatives.

较好的,它能够转录产生EV71毒株的全长感染性RNA或者其突变体。Preferably, it is capable of transcribing the full-length infectious RNA of the EV71 strain or a mutant thereof.

本发明还提供了一种疫苗或者病毒载体,它根据上述质粒制备;The present invention also provides a vaccine or viral vector, which is prepared according to the above-mentioned plasmid;

本发明提供了一种病毒颗粒,它由上述cDNA克隆或者质粒制备;The present invention provides a virus particle prepared from the above-mentioned cDNA clone or plasmid;

例如,减毒(live-attenuated)病毒颗粒,非感染复制性病毒(replicationcompetent non-infectious)颗粒及非复制性病毒(defective variants)颗粒;For example, live-attenuated virus particles, replicationcompetent non-infectious virus particles and non-replicative virus particles (defective variants);

所述的病毒可以通过免疫动物方法,分离纯化并获得抗EV71病毒抗体,也可以用于筛选人抗体库;或者,用于制备检测EV71病毒的试剂盒和各种细胞系、组织和动物感染模型。The virus can be isolated and purified to obtain anti-EV71 virus antibodies by immunizing animals, and can also be used to screen human antibody libraries; or, used to prepare kits for detecting EV71 virus and various cell lines, tissues and animal infection models .

上述细胞系、组织和动物感染模型可以用于筛选抗EV71病毒的药物。The above cell lines, tissues and animal infection models can be used to screen drugs against EV71 virus.

本发明还包括上述病毒载体和病毒颗粒的检测方法,制备方法;The present invention also includes the detection method and preparation method of the above-mentioned virus vector and virus particle;

例如,使用所述的病毒颗粒免疫动物并分离抗体,或者筛选人抗体库。For example, the viral particles are used to immunize animals and isolate antibodies, or to screen human antibody libraries.

另一方面,本发明提供了一种检测EV71的试剂盒,它含有上述cDNA或者病毒颗粒。In another aspect, the present invention provides a kit for detecting EV71, which contains the above-mentioned cDNA or virus particles.

本发明还提供了一种抗病毒EV71药物的制备方法,它使用上述cDNA或者病毒颗粒构建细胞或者动物模型,用于筛选抗病毒EV71的药物;或者,使用上述cDNA或者病毒颗粒构建细胞或者动物模型,用于筛选抗病毒EV71的药物。The present invention also provides a method for preparing an antiviral EV71 drug, which uses the above cDNA or virus particles to construct a cell or animal model for screening antiviral EV71 drugs; or, uses the above cDNA or virus particles to construct a cell or animal model , for the screening of antiviral EV71 drugs.

本发明的感染性克隆(核酸序列1)为一个由DNA序列构成的一个完整质粒(plasmid)。其中包含一个全长的EV71毒株(js1)的核酸序列(核酸序列2)及一个低拷贝质粒骨架序列(核酸序列3)。质粒(plasmid)是以共价键结合的闭合双链DNA(doublestranded DNA)。其中包含一条与mRNA序列一致的一条有义链(positive-sense strand)及一条与之互补的反义链或负义链(negative-sense strand)。The infectious clone of the present invention (nucleic acid sequence 1) is a complete plasmid consisting of a DNA sequence. It contains a full-length EV71 strain (js1) nucleic acid sequence (nucleic acid sequence 2) and a low-copy plasmid backbone sequence (nucleic acid sequence 3). Plasmids are closed double-stranded DNAs that are covalently bonded. It contains a positive-sense strand consistent with the mRNA sequence and a complementary antisense or negative-sense strand.

本发明的感染性克隆(核酸序列1)中所包含的EV71毒株(js1)的全长核酸序列(核酸序列2)包括病毒正链(positive sense)序列的5’末端的非翻译区(non-translatedregion,NTR)、一个开放阅读框(open reading frame,ORF)和3’末端非翻译区(3’-NTR)。在此感染性克隆中,病毒全长核酸序列5’末端含有一个T7启动子(TAA TAC GAC TCA CTA TAGG,SEQ ID NO 7)(图1A),可以在体外由商品化的T7转录试剂盒来转录病毒全长RNA;在病毒全长核酸序列3’末端含有一个30核苷酸长的polyA尾(AAAAA AAAAA AAAAA AAAAA AAAAAAAAAA,SEQ ID NO 8)(图1A)。The full-length nucleic acid sequence (nucleic acid sequence 2) of the EV71 strain (js1) contained in the infectious clone (nucleic acid sequence 1) of the present invention includes the untranslated region (non-translated region) at the 5' end of the viral positive sense sequence. -translated region, NTR), an open reading frame (open reading frame, ORF) and 3' terminal untranslated region (3'-NTR). In this infectious clone, the 5' end of the viral full-length nucleic acid sequence contains a T7 promoter (TAA TAC GAC TCA CTA TAGG, SEQ ID NO 7) (Figure 1A), which can be transcribed in vitro by a commercial T7 transcription kit Full-length viral RNA is transcribed; contains a 30-nucleotide polyA tail (AAAAA AAAAA AAAAA AAAAA AAAAAAAAAA, SEQ ID NO 8) at the 3' end of the viral full-length nucleic acid sequence (FIG. 1A).

临床分离毒株感染RD细胞,待细胞出现细胞病变时,抽提细胞的总RNA,利用EV71特异性引物(GCTAG CGCTtt tttttttt tttttttt ttttttt ttttt,SEQ ID NO 9)进行逆转录,然后利用逆转录(Superscript II逆转录酶)得到的cDNA进行PCR扩增。全长EV71基因组分4段扩增(图1A),扩增引物为F1(S:GACGC GGCCG CTAA TAC GACTC ACTATAG GTTAAAACAGC CTGT GGGT TGCAC CC,SEQ ID NO 10;As:GCACTG CACGT GGATGC AGAAC,SEQ ID NO11),F2(S:GACGCG GCCGCG TTCT GCAT CCAC GTGCA GTGC,SEQ ID NO 12;As:AAGTC GCGAGAGCT GTCTTC CC,SEQ ID NO 13),F3(S:GACGCG GCCGCG GGAA GACAG CTCTCG CGACTT,SEQID NO 14;As:AATTG TACAT CATG GTGC GATGG GTAGG,SEQ ID NO 15),F4(S:GACGC GGCCGCCCTAC CCATCG CACCATG ATGTAC AATT,SEQ ID NO 16;As:GCTAGC GCTtttttttttttttttt tttttttt ttttttGCT ATTCT GGTTAT AACAA ATTTA CCCCCA CCAG,SEQ ID NO17),扩增片段采用分步克隆的方法克隆到pANCR载体,得到最后的全长cDNA克隆,命名为pEV71-js1(图1A),RD cells were infected with clinically isolated virus strains. When the cells were cytopathic, the total RNA of the cells was extracted and reverse transcribed using EV71 specific primers (GCT AG CGCT tttttttttt tttttttt ttttttt ttttt, SEQ ID NO 9), and then reverse transcription (Superscript II reverse transcriptase) obtained cDNA was amplified by PCR. The full-length EV71 gene component was amplified in 4 segments (Figure 1A), and the amplification primer was F1(S: GAC GC GGCCG C TAA TAC GACTC ACTATAG GTTAAAACAGC CTGT GGGT TGCAC CC, SEQ ID NO 10; As: GCACTG CACGT GGATGC AGAAC, SEQ ID NO11), F2 (S: GAC GCG GCCGC G TTCT GCAT CCAC GTGCA GTGC, SEQ ID NO 12; As: AAGTC GCGAGAGCT GTCTTC CC, SEQ ID NO 13), F3 (S: GAC GCG GCCGC G GGAA GACAG CTCTCG CGACTT, SEQ ID NO 14; As: AATTG TACAT CATG GTGC GATGG GTAGG, SEQ ID NO 15), F4 (S: GAC GC GGCCGC CCTAC CCATCG CACCATG ATGTAC AATT, SEQ ID NO 16; As: GCT AGC GCT tttttttttttttttt tttttttt ttttttGCT ATTCT GGTTAT AACAA ATTTA CCCCCA CCAG, SEQ ID NO17), the amplified fragment was cloned into the pANCR vector by step-by-step cloning to obtain the final full-length cDNA clone, named pEV71-js1 (Fig. 1A),

此感染性克隆在体外经HindIII线性化后,由T7转录试剂盒来转录出含有病毒全长RNA及其3’末端的polyA尾。该体外产生的病毒RNA经电转或转染的方法导入到宿主细胞如Vero细胞后,病毒的RNA作为翻译模版,翻译其ORF,产生病毒多肽(蛋白序列4);该病毒多肽经加工形成病毒结构蛋白及非结构蛋白,起始整个病毒生活周期,产生子代病毒。After linearization with HindIII in vitro, the infectious clone was transcribed by the T7 transcription kit to contain the full-length viral RNA and its 3'-terminal polyA tail. After the in vitro-produced viral RNA is introduced into host cells such as Vero cells by electroporation or transfection, the viral RNA is used as a translation template to translate its ORF to produce a viral polypeptide (protein sequence 4); the viral polypeptide is processed to form a viral structure Proteins and non-structural proteins start the entire virus life cycle and produce progeny viruses.

由于基于编码的兼并性,通过改变密码子而不改变蛋白序列仍可以得到相同功能蛋白产物;本发明包括编码与“蛋白序列4”相同的其他核酸序列和感染性克隆。Due to degeneracy based on coding, the same functional protein product can still be obtained by changing codons without changing the protein sequence; the present invention includes other nucleic acid sequences and infectious clones encoding the same as "Protein Sequence 4".

本发明的感染性克隆(核酸序列1)所产生的病毒在细胞中表现出很强的复制能力(图2),可以用于感染体外培养的细胞系、神经组织、小鼠(图6)或猴等建立病毒感染的细胞模型及动物感染模型,用于药物的研发。The virus produced by the infectious clone (nucleic acid sequence 1) of the present invention exhibits strong replication ability in cells (Fig. 2), and can be used to infect in vitro cultured cell lines, neural tissues, mice (Fig. 6) or Monkeys and others established viral infection cell models and animal infection models for drug research and development.

通过对此感染性克隆(核酸序列1)进行改造,在病毒的特定区域(VP4蛋白编码区之前)插入报道基因,报道基因由病毒的IRES翻译起始。在报道基因的C末端添加额外氨基酸位点(AITTL)(图1B),此位点可被病毒的3C蛋白酶识别并切割,产生正常的VP4的N端。我们成功的在此感染性克隆(核酸序列1)中插入报道基因荧光素酶NanoLuc(Nluc)及荧光蛋白EGFP,分别构成带有Nluc的感染性克隆(核酸序列5)及带有EGFP的感染性克隆(核酸序列6)(图1B)。各报告基因通过融合PCR连接入pEV71-js1质粒(核酸序列1),分别命名为pEV71-js1-Nluc(核酸序列5)(图1B)及pEV71-js1-EGFP(核酸序列6)(图1C);此感染性克隆同上,在体外经HindIII线性化后,由T7转录试剂盒来转录出病毒全长RNA,体外转录的的病毒RNA经电转或转染的方法导入宿主细胞如Vero细胞后,可以起始病毒生活周期,产生子代病毒(图2)。病毒在复制过程中表达报道基因Nluc及EGFP。Nluc可以利用商品化的荧光素酶活性检测试剂盒进行检测(图5)。EGFP的表达可以利用荧光显微镜进行观察(图4)或利用流式细胞仪进行检测。产生的含有报道基因片段的子代病毒重新感染新细胞,在新细胞中可以有效复制。报道基因由于与病毒蛋白处于同一个开放阅读框,其表达水平反应病毒蛋白水平,亦可反应病毒复制水平。且含有报道基因的重组病毒在相当长的时间内连续传代报道基因无丢失(图4)。利用此含有报道基因的重组病毒,可以快速、方便的检测病毒复制及包装水平,可以用于研究病毒的生活周期、病毒-宿主相互作用、病毒的免疫学及抗病毒药物的开发等。This infectious clone (nucleic acid sequence 1) was engineered by inserting a reporter gene in a specific region of the virus (before the VP4 protein coding region), the reporter gene being translationally initiated by the IRES of the virus. An additional amino acid site (AITTL) was added to the C-terminus of the reporter gene (Fig. 1B), which was recognized and cleaved by the viral 3C protease, resulting in the normal N-terminus of VP4. We successfully inserted reporter gene luciferase NanoLuc (Nluc) and fluorescent protein EGFP into this infectious clone (nucleic acid sequence 1), which constituted an infectious clone with Nluc (nucleic acid sequence 5) and an infectious clone with EGFP, respectively. clone (nucleic acid sequence 6) (FIG. 1B). Each reporter gene was ligated into the pEV71-js1 plasmid (nucleic acid sequence 1) by fusion PCR and named as pEV71-js1-Nluc (nucleic acid sequence 5) (Fig. 1B) and pEV71-js1-EGFP (nucleic acid sequence 6) (Fig. 1C) ; This infectious clone is the same as above. After being linearized by HindIII in vitro, the full-length viral RNA is transcribed by the T7 transcription kit. The viral life cycle is initiated, producing progeny viruses (Figure 2). The virus expresses reporter genes Nluc and EGFP during replication. Nluc can be detected using a commercial luciferase activity detection kit (Figure 5). The expression of EGFP can be observed by fluorescence microscopy (Figure 4) or detected by flow cytometry. The resulting progeny virus containing the reporter gene segment re-infects new cells, where it can replicate efficiently. Since the reporter gene is in the same open reading frame as the viral protein, its expression level reflects the viral protein level and also the viral replication level. And the recombinant virus containing the reporter gene has no loss of the reporter gene in the continuous passage for a long period of time (Fig. 4). The recombinant virus containing the reporter gene can quickly and conveniently detect the level of virus replication and packaging, and can be used to study the life cycle of the virus, virus-host interaction, virus immunology and the development of antiviral drugs.

对此感染性克隆(核酸序列1)进行改造,参照肠道病毒属其他病毒,比如剔除病毒的结构蛋白VP4-VP3-VP2-VP1区域,可以构成病毒的亚基因组复制子(subgenomicreplicon)等复制非感染性病毒(replication competent non-infectious),该亚基因组复制子能进行病毒基因复制,但由于缺少病毒的结构蛋白不能包装出子代病毒。同时此亚基因组复制子RNA可以由表达的结构蛋白进行反式互补(trans complement),包装成重组亚病毒颗粒(recombinant subviral particles,RSPs)(Barclay,et al.J GenVirol.1998,79:1725-1734;Jia,et al.J Virol.1998,72:7972-7977),该亚病毒颗粒可以进行一轮感染,但由于基因组没有不编码结构蛋白,因此感染后不能再次病毒颗粒包装,是一种非复制性病毒(defective variants)颗粒。这些非复制性病毒颗粒可以作为一种型式的疫苗。This infectious clone (nucleic acid sequence 1) is modified with reference to other viruses belonging to the genus Enterovirus, such as deleting the VP4-VP3-VP2-VP1 region of the structural protein of the virus, which can constitute the subgenomic replicon of the virus and other non-replicating viruses. Infectious virus (replication competent non-infectious), the subgenomic replicon is capable of viral gene replication, but cannot package progeny virus due to the lack of viral structural proteins. At the same time, this subgenomic replicon RNA can be trans-complemented by the expressed structural protein and packaged into recombinant subviral particles (RSPs) (Barclay, et al. J GenVirol. 1998, 79:1725- 1734; Jia, et al. J Virol. 1998, 72: 7972-7977), this subviral particle can carry out a round of infection, but because the genome does not encode a structural protein, it cannot be packaged again after infection. Non-replicating viral particles (defective variants). These non-replicating viral particles can serve as a form of vaccine.

对此感染性克隆(核酸序列1)进行改造,可以构成减毒(live-attenuated)病毒,此减毒病毒可以作为疫苗。参考同为微小RNA病毒科的Polio病毒的减毒策略,在5’NTR上进行突变,可以构建减毒疫苗(Arita,et al.J Virol.2008,82:1787-1797)。This infectious clone (nucleic acid sequence 1) can be engineered to constitute a live-attenuated virus, which can be used as a vaccine. Referring to the attenuation strategy of Polio virus, which is also a picornaviridae, attenuated vaccine can be constructed by mutation on 5'NTR (Arita, et al. J Virol. 2008, 82: 1787-1797).

感染性克隆产生的病毒感染小鼠,建立方便、稳定的动物感染模型,本申请中,发现初始分离的毒株经过多于3次的传代后其感染新生小鼠后致死率下降,通过测序发现传代后的病毒其VP1的145位发生由E到G到突变,因此由分离的病毒进行感染建立老鼠模型一致性较差,利用感染性克隆产生病毒可以保证病毒序列不受细胞传代的影响。本申请利用上述感染性克隆得到的病毒感染不同品系(ICR,Balb/c,C57)的新生老鼠,均可以在9天内得到100%的死亡率(图6B),但携带有VP1的145G突变的病毒感染老鼠后不导致小鼠死亡(图6C),该动物模型可以方便的用于抗病毒药物以疫苗评价等。The virus produced by the infectious clone infects mice, and establishes a convenient and stable animal infection model. In the present application, it was found that the lethality of the initially isolated virus strain decreased after infecting neonatal mice after more than 3 passages. After passage, the 145 position of VP1 of the virus is mutated from E to G. Therefore, the mouse model established by the infection of the isolated virus has poor consistency. The use of infectious clone to generate virus can ensure that the virus sequence is not affected by cell passage. In this application, neonatal mice of different strains (ICR, Balb/c, C57) were infected with the virus obtained by the above infectious clone, and 100% mortality could be obtained within 9 days (Fig. 6B), but the mice carrying the 145G mutation of VP1 The virus did not lead to death of mice after infection (Fig. 6C). This animal model can be conveniently used for antiviral drugs and vaccine evaluation.

新型肠道病毒71型(Human enterovirus type71,EV71)为微小RNA病毒科属于微小病毒科(picornaviridae)中的肠病毒群(enterovirus)成员。EV71是在全球范围内引起儿童手足口病的主要病原体,它可以导致儿童罹患轻症和重症手足口病。病毒可感染中枢神经系统,引起中枢神经系统损伤,但其机制未明。目前无有效的治疗EV71的抗病毒药物。本发明通过分离一株临床EV71毒株,利用分子克隆,构建了稳定的病毒的全长cDNA克隆,通过体外转录RNA、转染Vero细胞证实所述的cDNA克隆来源的病毒RNA能产生EV71病毒;进一步,本申请构建了含有报道基因Gluc(Gaussia luciferase)及EGFP的重组病毒,并证实含有报道Gluc及EGFP的重组病毒具有感染宿主细胞并引起细胞病变的能力,利用感染性克隆来源的EV71病毒感染免疫健全的ICR、Bab/C及C57乳鼠,在10天内导致100%的被感染老鼠出现神经损伤症状而死亡。A new type of enterovirus type 71 (Human enterovirus type 71, EV71) is a member of the enterovirus group in the Picornaviridae family of Picornaviridae . EV71 is the main pathogen causing HFMD in children worldwide, and it can cause mild and severe HFMD in children. The virus can infect the central nervous system and cause damage to the central nervous system, but the mechanism is unknown. There are currently no effective antiviral drugs for EV71. In the present invention, a stable virus full-length cDNA clone is constructed by isolating a clinical EV71 strain by molecular cloning, and it is confirmed by in vitro transcription of RNA and transfection of Vero cells that the viral RNA derived from the cDNA clone can produce EV71 virus; Further, the application has constructed a recombinant virus containing reporter gene Gluc (Gaussia luciferase) and EGFP, and confirmed that the recombinant virus containing reporter Gluc and EGFP has the ability to infect host cells and cause cytopathic effects, and utilize the EV71 virus of infectious clone source to infect. The immune-competent ICR, Bab/C and C57 suckling mice caused 100% of the infected mice to develop neurological damage and die within 10 days.

本发明提供了稳定的、基于一株临床分离的EV71毒株的感染性cDNA克隆及其含有各类报道基因的衍生克隆、及以其为母本构建的各种突变克隆;以及利用这些克隆产生的各种重组病毒、亚单位病毒颗粒;以及利用这些克隆产生的各种重组病毒感染动物建立的动物模型;以及利用这些病毒或亚单位病毒颗粒用于疫苗的开发及诊断试剂的应用;以及利用此病毒作为基因治疗载体或表达载体。The present invention provides stable, infectious cDNA clones based on a clinically isolated EV71 strain, its derivative clones containing various reporter genes, and various mutant clones constructed therefrom; and the use of these clones to generate Various recombinant viruses and subunit virus particles produced by these clones; and animal models established by infecting animals with various recombinant viruses produced by these clones; and using these viruses or subunit virus particles for the development of vaccines and the application of diagnostic reagents; and the use of This virus is used as a gene therapy vector or an expression vector.

本发明的优点还有:The advantages of the present invention also include:

本发明包括利用这些克隆质粒为母本,通过分子生物学构建的各种重组病毒、亚单位病毒颗粒质粒。The present invention includes various recombinant viruses and subunit virus particle plasmids constructed by molecular biology using these cloned plasmids as mothers.

本发明还包括利用这些克隆可以产生的各种重组病毒、亚单位病毒颗粒;其含有上述cDNA。The present invention also includes various recombinant viruses, subunit virus particles that can be produced using these clones; which contain the cDNAs described above.

本发明还包括利用这些克隆可以产生的各种重组病毒构建的动物感染模型。The present invention also includes animal infection models constructed using the various recombinant viruses that can be produced by these clones.

本发明还包括利用这些病毒或亚单位病毒颗粒及动物模型用于疫苗的开发及诊断试剂。The present invention also includes the use of these viruses or subunit virus particles and animal models for vaccine development and diagnostic reagents.

本发明还包括利用这些病毒或亚单位病毒颗粒建立的动物模型用于疫苗的开发及抗病毒药物的开发。The present invention also includes animal models established by using these viruses or subunit virus particles for the development of vaccines and the development of antiviral drugs.

本发明还包括利用此病毒或亚病毒单位质粒作为基因治疗载体或表达载体质粒及利用这些质粒所产生的病毒或亚病毒颗粒。The present invention also includes the use of the viral or subviral unit plasmids as gene therapy vectors or expression vector plasmids and viral or subviral particles produced by using these plasmids.

本发明为EV71病毒感染的检测、预防、免疫提供了新的工具和途径,为利用此EV71毒株感染性克隆作为病毒载体进行基因治疗及疫苗开发提供了可能性。The present invention provides a new tool and approach for detection, prevention and immunization of EV71 virus infection, and provides a possibility for gene therapy and vaccine development by using the EV71 strain infectious clone as a viral vector.

附图说明Description of drawings

图1:EV71毒株js1的感染性cDNA克隆的构建,其中,Figure 1: Construction of an infectious cDNA clone of EV71 strain js1 in which,

(A)感染性克隆构建策略;寨卡病毒全基因组模式图,两端黑色柱子分别表示5’-NTR及3’-NTR;病毒结构蛋白区域及非结构蛋白区域如图所示;病毒全长序列分成4段分别进行扩增,其中第一段F1中含有T7序列,第四段F4含有由PCR引物引入的polyA30序列;合成的序列通过限制性内切酶依图所示依次连接入pACNR载体,得到全长克隆;(B)通过融合PCR,在VP4的N端同框融合Nluc或EGFP基因,Nluc或EGFP基因C端添加额外的氨基酸序列AITTL,便于被病毒蛋白酶切割从而产生正确的VP4的N端。(A) Infectious clone construction strategy; Zika virus whole genome schematic diagram, the black bars at both ends represent 5'-NTR and 3'-NTR respectively; the structural protein region and non-structural protein region of the virus are shown in the figure; the full length of the virus The sequence is divided into 4 segments and amplified respectively. The first segment F1 contains the T7 sequence, and the fourth segment F4 contains the polyA 30 sequence introduced by PCR primers; the synthesized sequences are sequentially connected to pACNR by restriction endonucleases as shown in the figure. (B) by fusion PCR, the Nluc or EGFP gene was fused in-frame at the N-terminus of VP4, and an additional amino acid sequence AITTL was added to the C-terminus of the Nluc or EGFP gene to facilitate cleavage by viral protease to produce correct VP4 the N-terminus of .

图2:EV71毒株js1的感染性cDNA克隆产生病毒的复制能力及感染能力,其中,Figure 2: Infectious cDNA clone of EV71 strain js1 produces virus replication and infectivity, wherein,

感染性克隆质粒作为模版,经体外转录成病毒RNA,病毒RNA通过电转导入到Vero细胞,收集上清病毒,在Vero细胞上利用噬斑实验进行滴度滴定,感染性克隆产生(Clone-WT)的噬斑同母本病毒(Parent)产生的噬斑的比较如图(上),相同滴度的感染性克隆产生的病毒同母本病毒再次感染Vero细胞(MOI=0.1),收集感染后不同时间(h.p.i)的细胞上清,利用噬斑实验对其进行滴定,得到两者的生长曲线如图(下),病毒滴度由PFU/ml表示。Infectious cloning plasmid was used as a template, transcribed into viral RNA in vitro, viral RNA was introduced into Vero cells by electroporation, supernatant virus was collected, titered by plaque assay on Vero cells, and infectious clones were generated (Clone-WT) The comparison between the plaques produced by the parent virus (Parent) and the plaques produced by the parent virus (Parent) is shown in the figure (top), the virus produced by the infectious clone of the same titer and the parent virus were re-infected with Vero cells (MOI=0.1), and the difference after infection was collected. Time (h.p.i) cell supernatant was titrated by plaque assay, and the growth curves of the two were obtained as shown in the figure (bottom), and the virus titer was represented by PFU/ml.

图3:含有报道基因Nluc及EGFP的重组病毒的产生,其中,Figure 3: Production of recombinant viruses containing reporter genes Nluc and EGFP, wherein,

(A)含有报道基因Nluc及EGFP的感染性克隆质粒,同不含报道基因的感染性克隆质粒,经体外转录成病毒RNA,病毒RNA通过电转导入到Vero细胞,收集上清病毒,在Vero细胞上利用噬斑实验进行滴度滴定,含各报道基因的重组病毒产生的噬斑同不含报道基因的病毒产生的噬斑的比较;(B)相同滴度的含各报道基因的病毒同不含报道基因的病毒再次感染Vero细胞(MOI=0.1),收集感染后不同天数的上清,利用噬斑实验对其进行滴定,得到的生长曲线;病毒滴度由PFU/ml表示。(A) Infectious cloned plasmids containing reporter genes Nluc and EGFP, and infectious cloned plasmids without reporter genes, were transcribed into viral RNA in vitro, and the viral RNA was introduced into Vero cells by electroporation, and the supernatant virus was collected, and the virus was collected in Vero cells. Titer titration was carried out using the plaque experiment, and the plaques produced by the recombinant viruses containing each reporter gene were compared with those produced by the virus without the reporter gene; (B) the same titer of the virus containing each reporter gene was different The virus containing the reporter gene was re-infected with Vero cells (MOI=0.1), and the supernatants of different days after infection were collected and titrated by plaque assay to obtain the growth curve; the virus titer was expressed by PFU/ml.

图4:含EGFP报道基因的重组病毒的稳定性,其中,Figure 4: Stability of recombinant virus containing EGFP reporter gene, wherein,

重组病毒EV71-EGFP感染的细胞上清以1:10稀释后,重新感染新的Vero细胞,感染二天后细胞用荧光显微镜观察及收集上清,再次以1:10稀释度重新感染新的Vero细胞(C+1),感染二天后同上细胞用荧光显微镜观察及收集上清进行再次感染;依次传代感染,观察感染细胞中EGFP的表达情况。The supernatant of cells infected with recombinant virus EV71-EGFP was diluted 1:10, and then re-infected with new Vero cells. After two days of infection, the cells were observed with a fluorescence microscope and the supernatant was collected, and the new Vero cells were re-infected with a 1:10 dilution. (C+1), two days after infection, the same cells were observed with a fluorescence microscope and the supernatant was collected for re-infection; the infection was successively subcultured, and the expression of EGFP in the infected cells was observed.

图5:含Nluc报道基因的重组病毒的产生Nluc的活性,其中,Figure 5: Nluc-producing activity of recombinant viruses containing the Nluc reporter gene, wherein,

含有报道基因Nluc的感染性克隆质粒同含有VP1 E145G及3C C147A突变的质粒,经体外转录成病毒RNA,病毒RNA通过电转导入到Vero细胞,电转后不同时间点收集细胞,检测细胞内Nluc活性,C147A为3C蛋白酶酶活性缺失突变。The infectious cloned plasmid containing the reporter gene Nluc and the plasmid containing the VP1 E145G and 3C C147A mutations were transcribed into viral RNA in vitro, and the viral RNA was introduced into Vero cells by electroporation. The cells were collected at different time points after electroporation, and the intracellular Nluc activity was detected. C147A is a deletion mutation of 3C protease enzymatic activity.

图6:EV71毒株js1的感染性cDNA克隆产生病毒感染小鼠构建动物感染模型,其中,Figure 6: Infectious cDNA clone of EV71 strain js1 produces virus-infected mice to construct an animal infection model, wherein,

(A)感染性cDNA克隆产生的病毒感染不同品系3日龄胎鼠(1.4×104pfu/只),感染后5天观察。(B)病毒感染后小鼠的生存曲线(n=5/group)。(C)感染性cDNA克隆(WT)及携带有VP1 E145G突变的克隆产生的病毒感染3日龄ICR小鼠,小鼠的生存曲线(n=5/group)。(A) Viruses generated from infectious cDNA clones infected different strains of 3-day-old fetal mice (1.4×10 4 pfu/mice) and observed 5 days after infection. (B) Survival curve of mice after virus infection (n=5/group). (C) Survival curves of 3-day-old ICR mice (n=5/group) generated by infectious cDNA clones (WT) and clones carrying the VP1 E145G mutation.

具体实施方式Detailed ways

本发明所用的方法均为常规的分子生物学方法,其中具体的操作细节不再赘述。The methods used in the present invention are all conventional molecular biology methods, and the specific operation details are not repeated here.

实施例1:EV71毒株js1的感染性cDNA克隆的构建Example 1: Construction of an infectious cDNA clone of EV71 strain js1

如图1A所示,从粪便标本中分离的病毒,经RD细胞培养,待细胞出现明显细胞病变时,提取总的细胞RNA,利用superscript II(Invitrogen)逆转录酶,以序列特异性引物(GCTAG CGCTttt tttttttttttt tttttttttt ttttt)进行逆转录,以得到的cDNA为模版,利用高保真酶super Fi(Invitrogen)分4段进行PCR扩增,扩增引物为F1(S:GACGC GGCCG CTAA TAC GACTC ACTATAG GTTAAA ACAGC CTGT GGGT TGCAC CC;As:GCACTG CACGT GGATGCAGAAC),F2(S:GACGCG GCCGCG TTCT GCAT CCAC GTGCA GTGC;As:AAGTC GCGA GAGCTGTCTTC CC),F3(S:GACGCG GCCGCG GGAA GACAG CTCTCG CGACTT;As:AATTG TACAT CATGGTGC GATGG GTAGG),F4(S:GACGC GGCCGCCCTAC CCATCG CACCATG ATGTAC AATT;As:GCTAGC GCTtttttttt tttttttt tttttttt ttttttGCT ATTCT GGTTAT AACAA ATTTA CCCCCACCAG),扩增的F4片段首先经限制性内切酶NotI/AfeI消化后,与经同样限制性内切酶消化的pANCR载体连接,得到pANCR-F4质粒,PCR扩增的F3片段利用NruI/BsrGI连接入pANCR-F4得到pANCR-F34质粒,PCR扩增的F2片段利用PmlI/NruI连接入pANCR-F34得到pANCR-F234,PCR扩增的F1片段利用NotI/PmlI连接入pANCR-F234得到最后的全长cDNA克隆,命名为pEV71-js1。As shown in Figure 1A, the virus isolated from fecal specimens was cultured in RD cells. When the cells showed obvious cytopathic changes, total cellular RNA was extracted, and the reverse transcriptase was superscript II (Invitrogen) and sequence-specific primers (GCT) were used. AG CGCT ttt tttttttttttt tttttttttt ttttt) was reverse transcribed, and the obtained cDNA was used as a template, and the high-fidelity enzyme super Fi (Invitrogen) was used for PCR amplification in 4 segments, and the amplification primer was F1 (S: GAC GC GGCCG C TAA TAC GACTC ACTATAG GTTAAA ACAGC CTGT GGGT TGCAC CC; As: GCACTG CACGT GGATGCAGAAC), F2 (S: GAC GCG GCCGC G TTCT GCAT CCAC GTGCA GTGC; As: AAGTC GCGA GAGCTGTCTTC CC), F3 (S: GAC GCG GCCGC G GGAA GACAG CTCTCG CGACTT; As: AATTG TACAT CATGGTGC GATGG GTAGG), F4 (S: GAC GC GGCCGC CCTAC CCATCG CACCATG ATGTAC AATT; As: GCT AGC GCT tttttttt tttttttt tttttttt ttttttGCT ATTCT GGTTAT AACAA ATTTA CCCCCACCAG), the amplified F4 fragment was first subjected to restriction endonuclease After digestion with NotI/AfeI, it was ligated with the pANCR vector digested with the same restriction enzymes to obtain the pANCR-F4 plasmid. The PCR-amplified F3 fragment was ligated into pANCR-F4 using NruI/BsrGI to obtain the pANCR-F34 plasmid, which was amplified by PCR. The F2 fragment was ligated into pANCR-F34 using PmlI/NruI to obtain pANCR-F234, and the PCR-amplified F1 fragment was ligated into pANCR-F234 using NotI/PmlI to obtain the final full-length cDNA clone, named pEV71-js1.

为构建带有报道基因EGFP的感染性克隆质粒(如图1B所示),利用融合PCR,融合三段序列,其中EGFP-F1为EV71 5UTR序列,PCR扩增引物为S:CCTGA CGTG TCGA CGCGG,SEQ IDNO 18,As:cctc gccct tgctcac CATcatatgG TTTAGCTGT GTTAAG GGTCAAGA,SEQ ID NO19,EGFP-F2为含有EGFP的片段,PCR扩增引物为S:TCTT GACC CTTAAC ACAGC TAA ACcatatgATG gtga gcaag ggcg agg,SEQ ID NO 20,As:CGCT GTGT AGACAC TTGCGA ACCAAGAGTGGTG ATCGC atgcat cttgtac agctcgt ccatgc cg,SEQ ID NO 21,EGFP-F3为含有包含VP4及VP2区域的片段,PCR扩增引物为S:cggca tggac gagct gtaca agatgc atGCGA TCACCACT CTTGG TTCGC AAGTG TCTA CACAG CG,SEQ ID NO 22;As:CTGC ACGT GGAT GCA GAACCC,SEQ ID NO 23,三个片段经融合PCR融合后,利用NotI/PmlI连接入pEV71-js1质粒,替换原质粒中的序列,得到pEV71-js1-EGFP质粒。In order to construct the infectious cloning plasmid with reporter gene EGFP (as shown in Figure 1B), use fusion PCR to fuse three sequences, wherein EGFP-F1 is EV71 5UTR sequence, and PCR amplification primers are S: CCTGA CGTG TCGA CGCGG, SEQ ID NO 18, As: cctc gccct tgctcac CATcatatgG TTTAGCTGT GTTAAG GGTCAAGA, SEQ ID NO 19, EGFP-F2 is a fragment containing EGFP, PCR amplification primer is S: TCTT GACC CTTAAC ACAGC TAA ACcatatgATG gtga gcaag ggcg agg, SEQ ID NO 20, As: CGCT GTGT AGACAC TTGCGA ACCAAGAGTGGTG ATCGC atgcat cttgtac agctcgt ccatgc cg, SEQ ID NO 21, EGFP-F3 is a fragment containing VP4 and VP2 regions, PCR amplification primers are S: cggca tggac gagct gtaca agatgc atGCGA TCACCACT CTTGG TTCGC AAGTG TCTA CACAG CG, SEQ ID NO 22; As: CTGC ACGT GGAT GCA GAACCC, SEQ ID NO 23, after the three fragments were fused by fusion PCR, they were connected into the pEV71-js1 plasmid using NotI/PmlI, and the sequence in the original plasmid was replaced to obtain pEV71 -js1-EGFP plasmid.

为构建带有报道基因Nluc的感染性克隆质粒(如图1B所示),利用融合PCR,融合两段序列,其中Nluc-F1为EV715UTR序列,PCR扩增引物为S:CTGC ACGT GGAT GCA GAA CCC,SEQ ID NO 24,As:gaaa tcttcg agtgtga agaccattct agaGTT TAGC TGTG TTA AGGG TCAAG,SEQ ID NO 25,EGFP-F2为含有Nluc的片段,PCR扩增引物为S:CTTG ACCC TTAAC ACAGCTAA ACtct agaat ggtctt cacac tcgaa gatttc,SEQ ID NO 26;As:CGCat gcatcg ccagaatgcgt tcgca,SEQ ID NO 27。两个片段经融合PCR融合后,利用NotI/NsiI连接入pEV71-js1-EGFP质粒,替换原质粒中的序列,得到pEV71-js1-Nluc质粒。In order to construct the infectious cloning plasmid with reporter gene Nluc (as shown in Figure 1B), use fusion PCR to fuse two sequences, wherein Nluc-F1 is the EV715UTR sequence, and the PCR amplification primer is S: CTGC ACGT GGAT GCA GAA CCC , SEQ ID NO 24, As: gaaa tcttcg agtgtga agaccattct agaGTT TAGC TGTG TTA AGGG TCAAG, SEQ ID NO 25, EGFP-F2 is a fragment containing Nluc, and the PCR amplification primer is S: CTTG ACCC TTAAC ACAGCTAA ACtct agaat ggtctt cacac tcgaa gatttc , SEQ ID NO 26; As: CGCat gcatcg ccagaatgcgt tcgca, SEQ ID NO 27. After the two fragments were fused by fusion PCR, they were ligated into the pEV71-js1-EGFP plasmid using NotI/NsiI, and the sequence in the original plasmid was replaced to obtain the pEV71-js1-Nluc plasmid.

实施例2:EV71毒株js1的感染性cDNA克隆产生病毒的复制能力及感染能力Example 2: Infectious cDNA clone of EV71 strain js1 produces virus replication ability and infectivity

感染性克隆质粒pEV71-js1用HindIII进行酶切,线性化,然后T7利用体外转录试剂盒(Ambion)。体外转录的RNA3g利用电转导的方法转入Vero细胞。电转后2天,待细胞出现病变,收集病毒上清,离心3000g,10min,然后经0.45m的滤膜过滤去掉细胞碎片。上清中的病毒利用噬斑实验进行滴定。感染性克隆质粒产生的病毒形成的噬斑与最初分离的母本病毒的噬斑比较如图2所示(上),两者噬斑的形态和大小无显著差异。相同滴度的感染性克隆产生的病毒同母本病毒再次感染Vero细胞(MOI=0.1),收集感染后不同时间的细胞上清,利用噬斑实验对其进行滴定(以PFU/ml表示),得到两者的生长曲线如图2所示(下),两者的生长曲线无显著差异。The infectious cloning plasmid pEV71-js1 was digested with HindIII, linearized, and then T7 using an in vitro transcription kit (Ambion). In vitro transcribed RNA3g was transferred into Vero cells by electrotransduction. 2 days after electroporation, when the cells became diseased, the virus supernatant was collected, centrifuged at 3000g for 10min, and then filtered through a 0.45m filter to remove cell debris. The virus in the supernatant was titered using a plaque assay. The plaques formed by the virus produced by the infectious cloning plasmid were compared with the plaques of the parent virus originally isolated as shown in Figure 2 (top), and there was no significant difference in the morphology and size of the plaques. The virus produced by the infectious clone of the same titer was re-infected with the parent virus in Vero cells (MOI=0.1), and the cell supernatants at different times after infection were collected and titrated by plaque assay (expressed in PFU/ml), The growth curves of the two are shown in Figure 2 (bottom), and there is no significant difference in the growth curves of the two.

实施例3:含有报道基因Nluc及EGFP的重组病毒的产生及其稳定性Embodiment 3: the production of the recombinant virus containing reporter gene Nluc and EGFP and its stability

含有报道基因Nluc及EGFP的感染性克隆质粒,同不含报道基因的感染性克隆质粒,同上经体外转录成病毒RNA,电转导入到Vero细胞,两天后收集细胞上清中的病毒,在Vero细胞上利用噬斑实验对其病毒滴度进行滴定。如图3A所示,含有报道基因EGFP及Nluc的病毒同不含报道基因的病毒其噬斑形态和大小类似。利用相同滴度的含各报道基因的病毒同不含报道基因的病毒再次感染Vero细胞(MOI=0.1),收集感染后不同天数的上清,利用噬斑实验对其进行滴定,得到的生长曲线,如图3B所示,携带有报道基因的病毒,相较于野生病毒,其生长周期表现为滞后,提示融合报道基因导致病毒复制周期的延缓。含有报道基因Nluc的感染性克隆质粒产出的病毒,其复制能力可以利用Nluc的底物(Promega),通过测定细胞内Nluc的活性进行判断。含有VP1 E145G及3C C147A突变的质粒,经体外转录成病毒RNA,病毒RNA通过电转导入到Vero细胞,不同时间测定细胞内Nluc活性,如图5所示,含有3C蛋白酶失活突变(C147A)的病毒RNA,其转染后,细胞内Nluc的活性在8小时后不再上升,其活性仅反应病毒RNA的起始翻译信号,但野生病毒活VP145G病毒RNA转染后,Nluc随时间延长表现为逐渐上升,表示正常病毒复制信号。为证明含有EGFP报道基因的病毒其报道基因的稳定性,我们利用EV71-EGFP病毒感染的细胞上清以1:10稀释后,重新感染新的Vero细胞,感染二天后细胞用荧光显微镜观察及收集上清,再次以1:10稀释度重新感染新的Vero细胞(C+1),感染二天后同上细胞用荧光显微镜观察及收集上清进行再次感染;依次传代感染,观察感染细胞中EGFP的表达情况,在连续传代至少6代后,EGFP基因仍然稳定。Infectious cloned plasmids containing reporter genes Nluc and EGFP, and infectious cloned plasmids without reporter genes, were transcribed into viral RNA in vitro as above, and electroporated into Vero cells. The virus titer was titrated by plaque assay. As shown in Figure 3A, the plaque morphology and size of the virus containing the reporter genes EGFP and Nluc were similar to the virus without the reporter gene. Vero cells were re-infected (MOI=0.1) with the same titer of the virus containing each reporter gene and the virus without the reporter gene, and the supernatants of different days after infection were collected and titrated by plaque assay. The obtained growth curve , as shown in Figure 3B, compared with the wild virus, the growth cycle of the virus carrying the reporter gene lagged behind, suggesting that the fusion of the reporter gene leads to the delay of the virus replication cycle. The replication ability of the virus produced by the infectious cloning plasmid containing the reporter gene Nluc can be judged by measuring the activity of intracellular Nluc using the Nluc substrate (Promega). The plasmids containing the VP1 E145G and 3C C147A mutations were transcribed into viral RNA in vitro, and the viral RNA was introduced into Vero cells by electroporation, and the intracellular Nluc activity was measured at different times. Viral RNA, after transfection, the activity of intracellular Nluc no longer rises after 8 hours, and its activity only reflects the initial translation signal of viral RNA, but after transfection of wild virus live VP145G viral RNA, Nluc over time appears as Gradually rise, indicating normal virus replication signal. In order to prove the stability of the reporter gene of the virus containing the EGFP reporter gene, we used the supernatant of EV71-EGFP virus-infected cells to be diluted 1:10, and then re-infected new Vero cells, and the cells were observed and collected with a fluorescence microscope two days after infection. The supernatant was re-infected with new Vero cells (C+1) at a dilution of 1:10. After two days of infection, the cells were observed with a fluorescence microscope and the supernatant was collected for re-infection; the infection was successively subcultured, and the expression of EGFP in the infected cells was observed. In some cases, the EGFP gene was still stable after at least 6 consecutive passages.

实施例4:EV71毒株js1的感染性cDNA克隆产生病毒感染小鼠构建动物感染模型Example 4: Infectious cDNA cloning of EV71 strain js1 produces virus-infected mice to construct an animal infection model

如图6A所示,感染性cDNA克隆产生的病毒感染不同品系3日龄胎鼠(1.4×104pfu/只),感染后5天观察小鼠,相比较不感染小鼠,感染有病毒的小鼠均表现为四肢瘫痪。各不同品系的小鼠感染后其生存曲线如图6B所示,在10天内,均达到100%死亡率。携带有VP1E145G突变的感染性克隆产生的病毒感染3日龄ICR小鼠后,不同于野生型小鼠,不导致小鼠的死亡,说明E145位点为病毒感染小鼠致死的决定性位点,也解释了传代病毒随传代次数的增加,其感染致死率降低的原因。As shown in Figure 6A, the virus produced by the infectious cDNA clone infected 3-day-old fetal mice of different strains (1.4×10 4 pfu/mice), and the mice were observed 5 days after infection. Compared with the uninfected mice, the mice infected with the virus The mice were all quadriplegic. The survival curves of different strains of mice after infection are shown in Figure 6B, and they all reached 100% mortality within 10 days. The virus produced by the infectious clone carrying the VP1E145G mutation infects 3-day-old ICR mice, which is different from wild-type mice and does not lead to the death of the mice, indicating that the E145 locus is the decisive locus for the lethality of virus-infected mice. Explains the reason why the infection lethality of the passaged virus decreases with the increase of the passage number.

序列表sequence listing

<110> 复旦大学<110> Fudan University

<120> 基于EV71毒株的感染性cDNA克隆及其应用<120> Infectious cDNA Cloning Based on EV71 Strain and Its Application

<130> 20190601<130> 20190601

<160> 27<160> 27

<170> SIPOSequenceListing 1.0<170> SIPOSequenceListing 1.0

<210> 1<210> 1

<211> 9446<211> 9446

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 1<400> 1

gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 60gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 60

tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 120tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 120

tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 180tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 180

atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 240atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 240

gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 300gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 300

aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 360aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 360

ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 420ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 420

tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 480tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 480

ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 540ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 540

ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 600ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 600

ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 660ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 660

gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 720gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 720

gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 780gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 780

gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct 840gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct 840

cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 900cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 900

acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 960acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 960

acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1020acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1020

tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1080tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1080

ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1140ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1140

ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 1200ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 1200

tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 1260tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 1260

aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 1320aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 1320

ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 1380ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 1380

ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 1440ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 1440

gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 1500gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 1500

gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 1560gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 1560

cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 1620cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 1620

actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 1680actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 1680

ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 1740ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 1740

tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 1800tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 1800

ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 1860ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 1860

agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 1920agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 1920

aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt gtcgacgcgg 1980aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt gtcgacgcgg 1980

ccgctaatac gactcactat aggttaaaac agcctgtggg ttgcacccac tcacagggcc 2040ccgctaatac gactcactat aggttaaaac agcctgtggg ttgcacccac tcacagggcc 2040

tactgggcgc aagcactctg gtacctcggt acctttgtgc gcctgtttta cacccccccc 2100tactgggcgc aagcactctg gtacctcggt acctttgtgc gcctgtttta cacccccccc 2100

ccaatgaaac ttagaagcaa taaaccacga tcaatagcag gcataacgct ccagttatgt 2160ccaatgaaac ttagaagcaa taaaccacga tcaatagcag gcataacgct ccagttatgt 2160

cttgatcaag cacttctgtt tccccggact gagtatcaat agactgctcg cgcggttgaa 2220cttgatcaag cacttctgtt tccccggact gagtatcaat agactgctcg cgcggttgaa 2220

ggagaaaacg ttcgttatcc ggctaactac ttcggaaaac ctagtaacac catgaaagtt 2280ggagaaaacg ttcgttatcc ggctaactac ttcggaaaac ctagtaacac catgaaagtt 2280

gcggagagct tcgttcagca ctcccccagt gtagatcagg tcgatgagtc accgcgttcc 2340gcggagagct tcgttcagca ctcccccagt gtagatcagg tcgatgagtc accgcgttcc 2340

ccacgggcga ccgtggcggt ggctgcgttg gcggcctgcc catggggtaa cccatggggc 2400ccacgggcga ccgtggcggt ggctgcgttg gcggcctgcc catggggtaa cccatggggc 2400

gctctaatac ggacatggtg tgaagagtct actgagctag ttggtagtcc tccggcccct 2460gctctaatac ggacatggtg tgaagagtct actgagctag ttggtagtcc tccggcccct 2460

gaatgcggct aatcccaact gcggagcaca cgcccacaag ccagcgggta gtgtgtcgta 2520gaatgcggct aatcccaact gcggagcaca cgcccacaag ccagcgggta gtgtgtcgta 2520

acgggtaact ctgcagcgga accgactact ttgggtgtcc gtgtttcctt ttatctttat 2580acgggtaact ctgcagcgga accgactact ttgggtgtcc gtgtttcctt ttatctttat 2580

attggctgct tatggtgaca attaaagaat tgttaccata tagctattgg attagccatc 2640attggctgct tatggtgaca attaaagaat tgttaccata tagctattgg attagccatc 2640

cggtgtgcaa cagagcaatt atttacctat ttattggttt tgtaccatta acctcgaatt 2700cggtgtgcaa cagagcaatt atttacctat ttattggttt tgtaccatta acctcgaatt 2700

ctgtgaccac ccttaattat atcttgaccc ttaacacagc taaacatggg ttcgcaagtg 2760ctgtgaccac ccttaattat atcttgaccc ttaacacagc taaacatggg ttcgcaagtg 2760

tctacacagc gctccggttc ttacgaaaac tcaaactcag ccactgaggg ttctaccata 2820tctacacagc gctccggttc ttacgaaaac tcaaactcag ccactgaggg ttctaccata 2820

aactacacca ccattaatta ctacaaagac tcctatgctg ccacagcagg caaacagagt 2880aactacacca ccattaatta ctacaaagac tcctatgctg ccacagcagg caaacagagt 2880

ctcaagcagg atccagacaa gtttgcaaat cctgttaaag acatattcac cgaaatggca 2940ctcaagcagg atccagacaa gtttgcaaat cctgttaaag acatattcac cgaaatggca 2940

gcgccactga agtccccatc cgctgaggca tgtggataca gtgatcgagt ggcgcaatta 3000gcgccactga agtccccatc cgctgaggca tgtggataca gtgatcgagt ggcgcaatta 3000

actattggca actccaccat cacgacgcaa gaagcggcta acatcatagt cggctatggt 3060actattggca actccaccat cacgacgcaa gaagcggcta acatcatagt cggctatggt 3060

gagtggcctt cctactgctc agattctgac gctacagcag tggataaacc aacgcgcccg 3120gagtggcctt cctactgctc agattctgac gctacagcag tggataaacc aacgcgcccg 3120

gatgtttcag tgaacaggtt ttacacattg gacactaaat tgtgggagaa atcgtccaag 3180gatgtttcag tgaacaggtt ttacacattg gacactaaat tgtgggagaa atcgtccaag 3180

ggatggtact ggaagttccc ggatgtgtta actgaaactg gggtttttgg gcaaaatgca 3240ggatggtact ggaagttccc ggatgtgtta actgaaactg gggtttttgg gcaaaatgca 3240

caattccact acctctaccg atcagggttc tgcatccacg tgcagtgcaa tgccagtaaa 3300caattccact acctctaccg atcagggttc tgcatccacg tgcagtgcaa tgccagtaaa 3300

ttccaccaag gagcactcct agtcgctgtc ctaccagagt atgtcattgg gacagtggca 3360ttccaccaag gagcactcct agtcgctgtc ctaccagagt atgtcattgg gacagtggca 3360

ggcggtacag ggacggaaga cacccacccc ccctacaagc agacccaacc cggcgccgat 3420ggcggtacag ggacggaaga cacccacccc ccctacaagc agacccaacc cggcgccgat 3420

ggtttcgagt tgcaacaccc gtacgtgctt gatgctggca tcccaatatc acagttaaca 3480ggtttcgagt tgcaacaccc gtacgtgctt gatgctggca tcccaatatc acagttaaca 3480

gtgtgcccac accagtggat taatttgagg accaacaatt gtgctacaat aatagtgcca 3540gtgtgcccac accagtggat taatttgagg accaacaatt gtgctacaat aatagtgcca 3540

tacattaacg cactgccttt tgattctgcc ttgaaccatt gcaactttgg cctgttagtt 3600tacattaacg cactgccttt tgattctgcc ttgaaccatt gcaactttgg cctgttagtt 3600

gtgcctatta gcccactaga ctacgaccaa ggagcaacgc cagtaatccc tataactatc 3660gtgcctatta gcccactaga ctacgaccaa ggagcaacgc cagtaatccc tataactatc 3660

acattggccc caatgtgctc tgaattcgca ggtcttaggc aggcagtcac gcaagggttc 3720acattggccc caatgtgctc tgaattcgca ggtcttaggc aggcagtcac gcaagggttc 3720

cccaccgagc taaaacctgg cacaaatcaa tttttaacca ccgatgatgg cgtctcagca 3780cccaccgagc taaaacctgg cacaaatcaa tttttaacca ccgatgatgg cgtctcagca 3780

cctattctac caaacttcca ccccaccccg tgtatccaca tacctggtga agttaggaac 3840cctattctac caaacttcca ccccaccccg tgtatccaca tacctggtga agttaggaac 3840

ttgctagagt tatgccaggt ggagaccatt ctggaggtta acaatgtgcc cacgaatgcc 3900ttgctagagt tatgccaggt ggagaccatt ctggaggtta acaatgtgcc cacgaatgcc 3900

actagcttaa tggagagact gcgcttcccg gtctcagcac aagcagggaa aggtgaactg 3960actagcttaa tggagagact gcgcttcccg gtctcagcac aagcagggaa aggtgaactg 3960

tgtgcggtgt ttagagccga tcctgggcga aatggaccat ggcaatccac cttactgggc 4020tgtgcggtgt ttagagccga tcctgggcga aatggaccat ggcaatccac cttactgggc 4020

cagttgtgcg ggtactacac ccaatggtca gggtcattgg aagtcacctt catgtttact 4080cagttgtgcg ggtactacac ccaatggtca gggtcattgg aagtcacctt catgtttact 4080

ggatccttca tggctaccgg caagatgctc atagcctata caccgccagg gggtcctctg 4140ggatccttca tggctaccgg caagatgctc atagcctata caccgccagg gggtcctctg 4140

cccaaggacc gggcgaccgc catgttgggc acgcacgtca tctgggattt tgggctgcaa 4200cccaaggacc gggcgaccgc catgttgggc acgcacgtca tctgggattt tgggctgcaa 4200

tcgtctgtta cccttgtaat accatggatc agtaacactc attatagagc acatgcccga 4260tcgtctgtta cccttgtaat accatggatc agtaacactc attatagagc acatgcccga 4260

gatggagtgt ttgactatta cactacaggg ttagtcagta tatggtacca gacaaattac 4320gatggagtgt ttgactatta cactacaggg ttagtcagta tatggtacca gacaaattac 4320

gtggttccaa tcggtgcgcc caacacagcc tatataatag cactagcggc agcccaaaag 4380gtggttccaa tcggtgcgcc caacacagcc tatataatag cactagcggc agcccaaaag 4380

aacttcacta tgaaattgtg caaggatgct agtgatatcc tgcagacggg caccatccag 4440aacttcacta tgaaattgtg caaggatgct agtgatatcc tgcagacggg caccatccag 4440

ggagataggg tggcagatgt aattgaaagt tccataggag atagcgtgag cagagccctc 4500ggagataggg tggcagatgt aattgaaagt tccataggag atagcgtgag cagagccctc 4500

actcacgctc taccagcacc cacaggccaa aacacacagg tgagcagtca tcgactggat 4560actcacgctc taccagcacc cacaggccaa aacacacagg tgagcagtca tcgactggat 4560

acaggcaagg ttccagcact ccaagctgct gaaattgggg catcatcaaa tgctagtgac 4620acaggcaagg ttccagcact ccaagctgct gaaattgggg catcatcaaa tgctagtgac 4620

gagagcatga ttgaaacacg ttgtgttctt aactcgcata gtacagctga gaccactctt 4680gagagcatga ttgaaacacg ttgtgttctt aactcgcata gtacagctga gaccactctt 4680

gatagtttct tcagtagggc aggattagtt ggagagatag atctccctct tgagggcaca 4740gatagtttct tcagtagggc aggattagtt ggagagatag atctccctct tgagggcaca 4740

actaacccaa atggttatgc caactgggac atagatataa caggttacgc gcaaatgcgt 4800actaacccaa atggttatgc caactgggac atagatataa caggttacgc gcaaatgcgt 4800

agaaaggtag agctattcac ctacatgcgt tttgatgcag agttcacttt tgttgcgtgc 4860agaaaggtag agctattcac ctacatgcgt tttgatgcag agttcacttt tgttgcgtgc 4860

acacccaccg gggaggttgt cccacaattg ctccaatata tgtttgtgcc acctggagcc 4920acacccaccg gggaggttgt cccacaattg ctccaatata tgtttgtgcc acctggagcc 4920

cctaagccag attctaggga atcccttgca tggcaaaccg ccaccaaccc ctcagttttt 4980cctaagccag attctaggga atcccttgca tggcaaaccg ccaccaaccc ctcagttttt 4980

gtcaagctgt cagaccctcc ggcgcaggtt tcagtgccat tcatgtcacc tgcgagtgct 5040gtcaagctgt cagaccctcc ggcgcaggtt tcagtgccat tcatgtcacc tgcgagtgct 5040

tatcaatggt tttatgacgg atatcccaca ttcggagaac acaaacagga gaaagacctt 5100tatcaatggt tttatgacgg atatcccaca ttcggagaac acaaacagga gaaagacctt 5100

gaatacgggg catgtcctaa taacatgatg ggtacattct cagtgcggac tgtggggacc 5160gaatacgggg catgtcctaa taacatgatg ggtacattct cagtgcggac tgtggggacc 5160

tccaagtcca agtacccttt agtggttagg atttacatga gaatgaagca cgtcagggcg 5220tccaagtcca agtacccttt agtggttagg atttacatga gaatgaagca cgtcagggcg 5220

tggatacctc gcccgatgcg caaccagaac tacctgttca aagccaaccc aaattatgct 5280tggatacctc gcccgatgcg caaccagaac tacctgttca aagccaaccc aaattatgct 5280

ggcaactcta ttaagccaac tggtgccagt cgcacagcga tcaccactct tgggaaattt 5340ggcaactcta ttaagccaac tggtgccagt cgcacagcga tcaccactct tgggaaattt 5340

ggacaacagt ctggggctat ttatgtgggc aactttagag tggtcaaccg acatcttgcc 5400ggacaacagt ctggggctat ttatgtgggc aactttagag tggtcaaccg acatcttgcc 5400

acccataatg attgggcaaa tcttgtttgg gaagacagct ctcgcgactt gctcgtgtca 5460acccataatg attgggcaaa tcttgtttgg gaagacagct ctcgcgactt gctcgtgtca 5460

tccaccactg cccaaggttg tgacacgatt gcccgttgcg attgccagac aggggtgtac 5520tccaccactg cccaaggttg tgacacgatt gcccgttgcg attgccagac aggggtgtac 5520

tactgtaact cgatgagaaa acactaccca gtcagttttt caaaacccag cctgatctat 5580tactgtaact cgatgagaaa acactaccca gtcagttttt caaaacccag cctgatctat 5580

gtagaggcta gcgagtatta cccagccagg taccaatcac atctcatgct cgcacagggt 5640gtagaggcta gcgagtatta cccagccagg taccaatcac atctcatgct cgcacagggt 5640

cactcggaac ctggtgattg cggtggtatc cttaggtgcc aacatggcgt catcggcata 5700cactcggaac ctggtgattg cggtggtatc cttaggtgcc aacatggcgt catcggcata 5700

gtgtctactg gtggcaatgg gctcgttggc tttgcagacg tcagagacct cttgtggtta 5760gtgtctactg gtggcaatgg gctcgttggc tttgcagacg tcagagacct cttgtggtta 5760

gatgaagaag ctatggaaca gggcgtgtcc gactacatta agggtctcgg agatgctttt 5820gatgaagaag ctatggaaca gggcgtgtcc gactacatta agggtctcgg agatgctttt 5820

ggaacaggct tcactgacgc agtctcaagg gaggttgaag ctctcaagaa ctatcttata 5880ggaacaggct tcactgacgc agtctcaagg gaggttgaag ctctcaagaa ctatcttata 5880

gggtctgaag gagcagttga gaaaattttg aaaaatctta ttaaactaat ctctgcactg 5940gggtctgaag gagcagttga gaaaattttg aaaaatctta ttaaactaat ctctgcactg 5940

gtgattgtga tcagaagtga ttacgacatg gttaccctca ctgcaacctt agcgctgata 6000gtgattgtga tcagaagtga ttacgacatg gttaccctca ctgcaacctt agcgctgata 6000

ggttgtcatg gcagtccttg ggcttggatt aaagccaaaa cagcctccat cttaggtatc 6060ggttgtcatg gcagtccttg ggcttggatt aaagccaaaa cagcctccat cttaggtatc 6060

cctatcgccc aaaagcagag cgcttcctgg ctcaagaagt tcaatgacat ggccaacgcc 6120cctatcgccc aaaagcagag cgcttcctgg ctcaagaagt tcaatgacat ggccaacgcc 6120

gctaaggggt tagagtgggt ttccaacaag atcagcaaat ttattgattg gcttaaggag 6180gctaaggggt tagagtgggt ttccaacaag atcagcaaat ttattgattg gcttaaggag 6180

aaaatagtac cagcagccag ggagaaggtt gaattcctaa ataacttgaa acagctgcca 6240aaaatagtac cagcagccag ggagaaggtt gaattcctaa ataacttgaa acagctgcca 6240

ctgctagaga atcagatctc gaacttggaa caatctgctg cttcacaaga ggaccttgaa 6300ctgctagaga atcagatctc gaacttggaa caatctgctg cttcacaaga ggaccttgaa 6300

gtcatgtttg ggaatgtgtc gtacctagct cacttctgtc gcaagtttca accgctatac 6360gtcatgtttg ggaatgtgtc gtacctagct cacttctgtc gcaagtttca accgctatac 6360

gccacggaag ctaaaagagt ctatgccctg gagaagagaa tgaataacta tatgcagttc 6420gccacggaag ctaaaagagt ctatgccctg gagaagagaa tgaataacta tatgcagttc 6420

aagagcaaac accgaattga acctgtatgt ctcattatta ggggctcacc aggcaccggg 6480aagagcaaac accgaattga acctgtatgt ctcattatta ggggctcacc aggcaccggg 6480

aagtctctag ccactggtat tattgctcga gcaatcgctg ataagtacca ctccagcgtg 6540aagtctctag ccactggtat tattgctcga gcaatcgctg ataagtacca ctccagcgtg 6540

tactcgctcc caccagaccc ggatcatttt gacggttaca agcaacaggt ggttacagtg 6600tactcgctcc caccagaccc ggatcatttt gacggttaca agcaacaggt ggttacagtg 6600

atggatgatt tgtgtcaaaa ccccgatggt aaggatatgt ccttattctg tcaaatggta 6660atggatgatt tgtgtcaaaa ccccgatggt aaggatatgt ccttattctg tcaaatggta 6660

tccaccgtag atttcattcc accaatggct tctctcgagg agaagggagt ttccttcacc 6720tccaccgtag atttcattcc accaatggct tctctcgagg agaagggagt ttccttcacc 6720

tctaagtttg tcatcgcatc cactaatgcc agtaatatca tagtaccaac agtgtctgat 6780tctaagtttg tcatcgcatc cactaatgcc agtaatatca tagtaccaac agtgtctgat 6780

tctgacgcta ttcgccgcag gttctacatg gactgtgaca ttgaagtgac agactcgtac 6840tctgacgcta ttcgccgcag gttctacatg gactgtgaca ttgaagtgac agactcgtac 6840

aaaacagatc taggtagact ggatgcaggg cgagccgcta aactgtgttc tgaaaataac 6900aaaacagatc taggtagact ggatgcaggg cgagccgcta aactgtgttc tgaaaataac 6900

actgcaaatt tcaaacgttg cagcccatta gtgtgtggga aagccatcca acttagagat 6960actgcaaatt tcaaacgttg cagcccatta gtgtgtggga aagccatcca acttagagat 6960

agaaagtcta aagtcagata cagtgtggat acggtggttt cagaacttat tagggaatac 7020agaaagtcta aagtcagata cagtgtggat acggtggttt cagaacttat tagggaatac 7020

agcaataggt ccgccattgg taacacaatc gaggctcttt tccaaggtcc acccaagttc 7080agcaataggt ccgccattgg taacacaatc gaggctcttt tccaaggtcc acccaagttc 7080

aggccaatta ggattagcct tgaagaaaaa ccagccccag acgctattag cgatctcctt 7140aggccaatta ggattagcct tgaagaaaaa ccagccccag acgctattag cgatctcctt 7140

gctagtgtag atagtgaaga agtgcgccag tactgcaggg atcaaggctg gattattcct 7200gctagtgtag atagtgaaga agtgcgccag tactgcaggg atcaaggctg gattattcct 7200

gaagctccca ccaatgtgga gcggcacctt aatagagcgg tgctcgtcat gcaatccatc 7260gaagctccca ccaatgtgga gcggcacctt aatagagcgg tgctcgtcat gcaatccatc 7260

accacagtag tggcggttgt ttcgttggtg tacgtcatct acaagctctt tgcagggttt 7320accacagtag tggcggttgt ttcgttggtg tacgtcatct acaagctctt tgcagggttt 7320

cagggtgcat attctggtgc tcctaagcaa gtgcttaaga aacctgctct tcgcacagca 7380cagggtgcat attctggtgc tcctaagcaa gtgcttaaga aacctgctct tcgcacagca 7380

acagtgcagg gtccgagcct tgactttgct ctctccctac tgagaaggaa catcaggcag 7440acagtgcagg gtccgagcct tgactttgct ctctccctac tgagaaggaa catcaggcag 7440

gtccaaacag accaagggca tttcaccatg ttgggtgtta gggatcgctt agcagtcctc 7500gtccaaacag accaagggca tttcaccatg ttgggtgtta gggatcgctt agcagtcctc 7500

ccacgccact cacaacctgg caaaaccatt tggattgagc acaaactcgt gaacgtcctt 7560ccacgccact cacaacctgg caaaaccatt tggattgagc acaaactcgt gaacgtcctt 7560

gatgcagttg aactggtgga tgagcaagga gtcaacctgg aattaaccct catcactctt 7620gatgcagttg aactggtgga tgagcaagga gtcaacctgg aattaaccct catcactctt 7620

gacaccaacg agaagtttag ggatatcacc aaattcatcc cagaaaatat cagcactgct 7680gacaccaacg agaagtttag ggatatcacc aaattcatcc cagaaaatat cagcactgct 7680

agcgatgcca ccctagtgat caacacggag cacatgccgt caatgtttgt cccggtgggt 7740agcgatgcca ccctagtgat caacacggag cacatgccgt caatgtttgt cccggtgggt 7740

gacgttgtgc agtatggctt tttgaatctc agtggcaagc ctacccatcg caccatgatg 7800gacgttgtgc agtatggctt tttgaatctc agtggcaagc ctacccatcg caccatgatg 7800

tacaattttc ctactaaagc aggacagtgt ggaggagtgg tgacatctgt tgggaaggtt 7860tacaattttc ctactaaagc aggacagtgt ggaggagtgg tgacatctgt tgggaaggtt 7860

gtcggtattc acattggtgg caatggcaga caaggttttt gcgcaggcct caaaaggagt 7920gtcggtattc acattggtgg caatggcaga caaggttttt gcgcaggcct caaaaggagt 7920

tactttgcta gtgaacaagg agagatccag tgggttaagc ccaataaaga aactggaaga 7980tactttgcta gtgaacaagg agagatccag tgggttaagc ccaataaaga aactggaaga 7980

ctcaacatca atggaccaac ccgcaccaag ttagaaccta gtgtattcca tgacatcttc 8040ctcaacatca atggaccaac ccgcaccaag ttagaaccta gtgtattcca tgacatcttc 8040

gagggaaata aggaaccagc tgtcttgcac agtaaagacc cccgacttga ggtagatttt 8100gagggaaata aggaaccagc tgtcttgcac agtaaagacc cccgacttga ggtagatttt 8100

gaacaggccc tgttctctaa gtatgtggga aacacactac atgagcctga cgagtacatc 8160gaacaggccc tgttctctaa gtatgtggga aacacactac atgagcctga cgagtacatc 8160

aaagaggcag ctctacatta tgcaaaccaa ttaaagcaac tagaaatcaa tacctctcaa 8220aaagaggcag ctctacatta tgcaaaccaa ttaaagcaac tagaaatcaa tacctctcaa 8220

atgagcatgg aggaggcctg ctatggtact gagaatcttg aggctattga tcttcacact 8280atgagcatgg aggaggcctg ctatggtact gagaatcttg aggctattga tcttcacact 8280

agtgcaggtt acccctatag tgccctaggg ataaagaaaa gagacatctt agaccctacc 8340agtgcaggtt acccctatag tgccctaggg ataaagaaaa gagacatctt agaccctacc 8340

accagggacg tgagtagaat gaagttctac atggacaagt atggtcttga tcttccctac 8400accagggacg tgagtagaat gaagttctac atggacaagt atggtcttga tcttccctac 8400

tccacttatg tcaaggacga gctacgctcg attgataaaa tcaagaaagg gaagtcccgc 8460tccacttatg tcaaggacga gctacgctcg attgataaaa tcaagaaagg gaagtcccgc 8460

ctgatcgagg ccagtagtct aaatgattca gtgtacctca gaatggcttt cgggcatttg 8520ctgatcgagg ccagtagtct aaatgattca gtgtacctca gaatggcttt cgggcatttg 8520

tatgaggctt tccacgcaaa tcctgggacg ataactggat cggccgtggg gtgtaaccct 8580tatgaggctt tccacgcaaa tcctgggacg ataactggat cggccgtggg gtgtaaccct 8580

gacacattct ggagcaagct gccaattttg ctccctggtt cactctttgc ctttgactac 8640gacacattct ggagcaagct gccaattttg ctccctggtt cactctttgc ctttgactac 8640

tcaggctatg atgccagcct tagccctgtc tggttcagag cattagaatt ggttcttagg 8700tcaggctatg atgccagcct tagccctgtc tggttcagag cattagaatt ggttcttagg 8700

gagatagggt atagtgaaga ggcaatctca ctcattgagg gaatcaacca cacacatcat 8760gagatagggt atagtgaaga ggcaatctca ctcattgagg gaatcaacca cacacatcat 8760

gtgtatcgta ataagaccta ttgcgtgctt ggtgggatgc cctcaggctg ttcaggaaca 8820gtgtatcgta ataagaccta ttgcgtgctt ggtgggatgc cctcaggctg ttcaggaaca 8820

tccatcttca actcaatgat caacaacatt attatcagag cactgctcat aaaaacattt 8880tccatcttca actcaatgat caacaacatt attatcagag cactgctcat aaaaacattt 8880

aagggcattg atttggatga actcaacatg gtcgcttatg gagacgatgt gctcgctagc 8940aagggcattg atttggatga actcaacatg gtcgcttatg gagacgatgt gctcgctagc 8940

tatcccttcc caattgattg cttggaacta gcaaagactg gtaaggagta tggtctgacc 9000tatcccttcc caattgattg cttggaacta gcaaagactg gtaaggagta tggtctgacc 9000

atgacccctg ctgataaatc tccttgcttt aatgaggtca attggggtaa tgcgaccttc 9060atgacccctg ctgataaatc tccttgcttt aatgaggtca attggggtaa tgcgaccttc 9060

ctcaaaaggg gctttttgcc cgatgaacag tttccatttt tgattcaccc tactatgcca 9120ctcaaaaggg gctttttgcc cgatgaacag tttccatttt tgattcaccc tactatgcca 9120

atgagggaga tccatgagtc cattcgatgg accaaggacg cacggaacac tcaagatcat 9180atgagggaga tccatgagtc cattcgatgg accaaggacg cacggaacac tcaagatcat 9180

gtgcggtcct tgtgcctcct agcatggcat aatggtaagc aagaatacga gaagtttgtg 9240gtgcggtcct tgtgcctcct agcatggcat aatggtaagc aagaatacga gaagtttgtg 9240

agcacaatta ggtctgtccc agtagggaga gcgttggcta ttccaaatta tgaaaatctt 9300agcacaatta ggtctgtccc agtagggaga gcgttggcta ttccaaatta tgaaaatctt 9300

agacgaaatt ggctcgagtt attttagagg ttatacacac ctcaacccca ccagaaatct 9360agacgaaatt ggctcgagtt attttagagg ttatacacac ctcaacccca ccagaaatct 9360

ggtcgtgaat gtgactggtg ggggtaaatt tgttataacc agaatagcaa aaaaaaaaaa 9420ggtcgtgaat gtgactggtg ggggtaaatt tgttataacc agaatagcaa aaaaaaaaaa 9420

aaaaaaaaaa aaaaaaaaaa gcttat 9446aaaaaaaaaa aaaaaaaaaa gcttat 9446

<210> 2<210> 2

<211> 7405<211> 7405

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 2<400> 2

ttaaaacagc ctgtgggttg cacccactca cagggcctac tgggcgcaag cactctggta 60ttaaaacagc ctgtgggttg cacccactca cagggcctac tgggcgcaag cactctggta 60

cctcggtacc tttgtgcgcc tgttttacac ccccccccca atgaaactta gaagcaataa 120cctcggtacc tttgtgcgcc tgttttacac ccccccccca atgaaactta gaagcaataa 120

accacgatca atagcaggca taacgctcca gttatgtctt gatcaagcac ttctgtttcc 180accacgatca atagcaggca taacgctcca gttatgtctt gatcaagcac ttctgtttcc 180

ccggactgag tatcaataga ctgctcgcgc ggttgaagga gaaaacgttc gttatccggc 240ccggactgag tatcaataga ctgctcgcgc ggttgaagga gaaaacgttc gttatccggc 240

taactacttc ggaaaaccta gtaacaccat gaaagttgcg gagagcttcg ttcagcactc 300taactacttc ggaaaaccta gtaacaccat gaaagttgcg gagagcttcg ttcagcactc 300

ccccagtgta gatcaggtcg atgagtcacc gcgttcccca cgggcgaccg tggcggtggc 360ccccagtgta gatcaggtcg atgagtcacc gcgttcccca cgggcgaccg tggcggtggc 360

tgcgttggcg gcctgcccat ggggtaaccc atggggcgct ctaatacgga catggtgtga 420tgcgttggcg gcctgcccat ggggtaaccc atggggcgct ctaatacgga catggtgtga 420

agagtctact gagctagttg gtagtcctcc ggcccctgaa tgcggctaat cccaactgcg 480agagtctact gagctagttg gtagtcctcc ggcccctgaa tgcggctaat cccaactgcg 480

gagcacacgc ccacaagcca gcgggtagtg tgtcgtaacg ggtaactctg cagcggaacc 540gagcacacgc ccacaagcca gcgggtagtg tgtcgtaacg ggtaactctg cagcggaacc 540

gactactttg ggtgtccgtg tttcctttta tctttatatt ggctgcttat ggtgacaatt 600gactactttg ggtgtccgtg tttcctttta tctttatatt ggctgcttat ggtgacaatt 600

aaagaattgt taccatatag ctattggatt agccatccgg tgtgcaacag agcaattatt 660aaagaattgt taccatatag ctattggatt agccatccgg tgtgcaacag agcaattatt 660

tacctattta ttggttttgt accattaacc tcgaattctg tgaccaccct taattatatc 720tacctattta ttggttttgt accattaacc tcgaattctg tgaccaccct taattatatc 720

ttgaccctta acacagctaa acatgggttc gcaagtgtct acacagcgct ccggttctta 780ttgaccctta acacagctaa acatgggttc gcaagtgtct acacagcgct ccggttctta 780

cgaaaactca aactcagcca ctgagggttc taccataaac tacaccacca ttaattacta 840cgaaaactca aactcagcca ctgagggttc taccataaac tacaccacca ttaattacta 840

caaagactcc tatgctgcca cagcaggcaa acagagtctc aagcaggatc cagacaagtt 900caaagactcc tatgctgcca cagcaggcaa acagagtctc aagcaggatc cagacaagtt 900

tgcaaatcct gttaaagaca tattcaccga aatggcagcg ccactgaagt ccccatccgc 960tgcaaatcct gttaaagaca tattcaccga aatggcagcg ccactgaagt ccccatccgc 960

tgaggcatgt ggatacagtg atcgagtggc gcaattaact attggcaact ccaccatcac 1020tgaggcatgt ggatacagtg atcgagtggc gcaattaact attggcaact ccaccatcac 1020

gacgcaagaa gcggctaaca tcatagtcgg ctatggtgag tggccttcct actgctcaga 1080gacgcaagaa gcggctaaca tcatagtcgg ctatggtgag tggccttcct actgctcaga 1080

ttctgacgct acagcagtgg ataaaccaac gcgcccggat gtttcagtga acaggtttta 1140ttctgacgct acagcagtgg ataaaccaac gcgcccggat gtttcagtga acaggtttta 1140

cacattggac actaaattgt gggagaaatc gtccaaggga tggtactgga agttcccgga 1200cacattggac actaaattgt gggagaaatc gtccaaggga tggtactgga agttcccgga 1200

tgtgttaact gaaactgggg tttttgggca aaatgcacaa ttccactacc tctaccgatc 1260tgtgttaact gaaactgggg tttttgggca aaatgcacaa ttccactacc tctaccgatc 1260

agggttctgc atccacgtgc agtgcaatgc cagtaaattc caccaaggag cactcctagt 1320agggttctgc atccacgtgc agtgcaatgc cagtaaattc caccaaggag cactcctagt 1320

cgctgtccta ccagagtatg tcattgggac agtggcaggc ggtacaggga cggaagacac 1380cgctgtccta ccagagtatg tcattgggac agtggcaggc ggtacaggga cggaagacac 1380

ccaccccccc tacaagcaga cccaacccgg cgccgatggt ttcgagttgc aacacccgta 1440ccacccccccc tacaagcaga cccaacccgg cgccgatggt ttcgagttgc aacacccgta 1440

cgtgcttgat gctggcatcc caatatcaca gttaacagtg tgcccacacc agtggattaa 1500cgtgcttgat gctggcatcc caatatcaca gttaacagtg tgcccacacc agtggattaa 1500

tttgaggacc aacaattgtg ctacaataat agtgccatac attaacgcac tgccttttga 1560tttgaggacc aacaattgtg ctacaataat agtgccatac attaacgcac tgccttttga 1560

ttctgccttg aaccattgca actttggcct gttagttgtg cctattagcc cactagacta 1620ttctgccttg aaccattgca actttggcct gttagttgtg cctattagcc cactagacta 1620

cgaccaagga gcaacgccag taatccctat aactatcaca ttggccccaa tgtgctctga 1680cgaccaagga gcaacgccag taatccctat aactatcaca ttggccccaa tgtgctctga 1680

attcgcaggt cttaggcagg cagtcacgca agggttcccc accgagctaa aacctggcac 1740attcgcaggt ctaggcagg cagtcacgca agggttcccc accgagctaa aacctggcac 1740

aaatcaattt ttaaccaccg atgatggcgt ctcagcacct attctaccaa acttccaccc 1800aaatcaattt ttaaccaccg atgatggcgt ctcagcacct attctaccaa acttccaccc 1800

caccccgtgt atccacatac ctggtgaagt taggaacttg ctagagttat gccaggtgga 1860caccccgtgt atccacatac ctggtgaagt taggaacttg ctagagttat gccaggtgga 1860

gaccattctg gaggttaaca atgtgcccac gaatgccact agcttaatgg agagactgcg 1920gaccattctg gaggttaaca atgtgcccac gaatgccact agcttaatgg agagactgcg 1920

cttcccggtc tcagcacaag cagggaaagg tgaactgtgt gcggtgttta gagccgatcc 1980cttcccggtc tcagcacaag cagggaaagg tgaactgtgt gcggtgttta gagccgatcc 1980

tgggcgaaat ggaccatggc aatccacctt actgggccag ttgtgcgggt actacaccca 2040tgggcgaaat ggaccatggc aatccacctt actgggccag ttgtgcgggt actacaccca 2040

atggtcaggg tcattggaag tcaccttcat gtttactgga tccttcatgg ctaccggcaa 2100atggtcaggg tcattggaag tcaccttcat gtttactgga tccttcatgg ctaccggcaa 2100

gatgctcata gcctatacac cgccaggggg tcctctgccc aaggaccggg cgaccgccat 2160gatgctcata gcctatacac cgccaggggg tcctctgccc aaggaccggg cgaccgccat 2160

gttgggcacg cacgtcatct gggattttgg gctgcaatcg tctgttaccc ttgtaatacc 2220gttgggcacg cacgtcatct gggattttgg gctgcaatcg tctgttaccc ttgtaatacc 2220

atggatcagt aacactcatt atagagcaca tgcccgagat ggagtgtttg actattacac 2280atggatcagt aacactcatt atagagcaca tgcccgagat ggagtgtttg actattacac 2280

tacagggtta gtcagtatat ggtaccagac aaattacgtg gttccaatcg gtgcgcccaa 2340tacagggtta gtcagtatat ggtaccagac aaattacgtg gttccaatcg gtgcgcccaa 2340

cacagcctat ataatagcac tagcggcagc ccaaaagaac ttcactatga aattgtgcaa 2400cacagcctat ataatagcac tagcggcagc ccaaaagaac ttcactatga aattgtgcaa 2400

ggatgctagt gatatcctgc agacgggcac catccaggga gatagggtgg cagatgtaat 2460ggatgctagt gatatcctgc agacgggcac catccaggga gatagggtgg cagatgtaat 2460

tgaaagttcc ataggagata gcgtgagcag agccctcact cacgctctac cagcacccac 2520tgaaagttcc ataggagata gcgtgagcag agccctcact cacgctctac cagcacccac 2520

aggccaaaac acacaggtga gcagtcatcg actggataca ggcaaggttc cagcactcca 2580aggccaaaac acacaggtga gcagtcatcg actggataca ggcaaggttc cagcactcca 2580

agctgctgaa attggggcat catcaaatgc tagtgacgag agcatgattg aaacacgttg 2640agctgctgaa attggggcat catcaaatgc tagtgacgag agcatgattg aaacacgttg 2640

tgttcttaac tcgcatagta cagctgagac cactcttgat agtttcttca gtagggcagg 2700tgttcttaac tcgcatagta cagctgagac cactcttgat agtttcttca gtagggcagg 2700

attagttgga gagatagatc tccctcttga gggcacaact aacccaaatg gttatgccaa 2760attagttgga gagatagatc tccctcttga gggcacaact aacccaaatg gttatgccaa 2760

ctgggacata gatataacag gttacgcgca aatgcgtaga aaggtagagc tattcaccta 2820ctgggacata gatataacag gttacgcgca aatgcgtaga aaggtagagc tattcaccta 2820

catgcgtttt gatgcagagt tcacttttgt tgcgtgcaca cccaccgggg aggttgtccc 2880catgcgtttt gatgcagagt tcacttttgt tgcgtgcaca cccaccgggg aggttgtccc 2880

acaattgctc caatatatgt ttgtgccacc tggagcccct aagccagatt ctagggaatc 2940acaattgctc caatatatgt ttgtgccacc tggagcccct aagccagatt ctagggaatc 2940

ccttgcatgg caaaccgcca ccaacccctc agtttttgtc aagctgtcag accctccggc 3000ccttgcatgg caaaccgcca ccaacccctc agtttttgtc aagctgtcag accctccggc 3000

gcaggtttca gtgccattca tgtcacctgc gagtgcttat caatggtttt atgacggata 3060gcaggtttca gtgccattca tgtcacctgc gagtgcttat caatggtttt atgacggata 3060

tcccacattc ggagaacaca aacaggagaa agaccttgaa tacggggcat gtcctaataa 3120tcccacattc ggagaacaca aacaggagaa agaccttgaa tacggggcat gtcctaataa 3120

catgatgggt acattctcag tgcggactgt ggggacctcc aagtccaagt accctttagt 3180catgatgggt acattctcag tgcggactgt ggggacctcc aagtccaagt accctttagt 3180

ggttaggatt tacatgagaa tgaagcacgt cagggcgtgg atacctcgcc cgatgcgcaa 3240ggttaggatt tacatgagaa tgaagcacgt cagggcgtgg atacctcgcc cgatgcgcaa 3240

ccagaactac ctgttcaaag ccaacccaaa ttatgctggc aactctatta agccaactgg 3300ccagaactac ctgttcaaag ccaacccaaa ttatgctggc aactctatta agccaactgg 3300

tgccagtcgc acagcgatca ccactcttgg gaaatttgga caacagtctg gggctattta 3360tgccagtcgc acagcgatca ccactcttgg gaaatttgga caacagtctg gggctattta 3360

tgtgggcaac tttagagtgg tcaaccgaca tcttgccacc cataatgatt gggcaaatct 3420tgtgggcaac tttagagtgg tcaaccgaca tcttgccacc cataatgatt gggcaaatct 3420

tgtttgggaa gacagctctc gcgacttgct cgtgtcatcc accactgccc aaggttgtga 3480tgtttgggaa gacagctctc gcgacttgct cgtgtcatcc accactgccc aaggttgtga 3480

cacgattgcc cgttgcgatt gccagacagg ggtgtactac tgtaactcga tgagaaaaca 3540cacgattgcc cgttgcgatt gccagacagg ggtgtactac tgtaactcga tgagaaaaca 3540

ctacccagtc agtttttcaa aacccagcct gatctatgta gaggctagcg agtattaccc 3600ctacccagtc agtttttcaa aacccagcct gatctatgta gaggctagcg agtattaccc 3600

agccaggtac caatcacatc tcatgctcgc acagggtcac tcggaacctg gtgattgcgg 3660agccaggtac caatcacatc tcatgctcgc acagggtcac tcggaacctg gtgattgcgg 3660

tggtatcctt aggtgccaac atggcgtcat cggcatagtg tctactggtg gcaatgggct 3720tggtatcctt aggtgccaac atggcgtcat cggcatagtg tctactggtg gcaatgggct 3720

cgttggcttt gcagacgtca gagacctctt gtggttagat gaagaagcta tggaacaggg 3780cgttggcttt gcagacgtca gagacctctt gtggttagat gaagaagcta tggaacaggg 3780

cgtgtccgac tacattaagg gtctcggaga tgcttttgga acaggcttca ctgacgcagt 3840cgtgtccgac tacattaagg gtctcggaga tgcttttgga acaggcttca ctgacgcagt 3840

ctcaagggag gttgaagctc tcaagaacta tcttataggg tctgaaggag cagttgagaa 3900ctcaagggag gttgaagctc tcaagaacta tcttataggg tctgaaggag cagttgagaa 3900

aattttgaaa aatcttatta aactaatctc tgcactggtg attgtgatca gaagtgatta 3960aattttgaaa aatcttatta aactaatctc tgcactggtg attgtgatca gaagtgatta 3960

cgacatggtt accctcactg caaccttagc gctgataggt tgtcatggca gtccttgggc 4020cgacatggtt accctcactg caaccttagc gctgataggt tgtcatggca gtccttgggc 4020

ttggattaaa gccaaaacag cctccatctt aggtatccct atcgcccaaa agcagagcgc 4080ttggattaaa gccaaaacag cctccatctt aggtatccct atcgcccaaa agcagagcgc 4080

ttcctggctc aagaagttca atgacatggc caacgccgct aaggggttag agtgggtttc 4140ttcctggctc aagaagttca atgacatggc caacgccgct aaggggttag agtgggtttc 4140

caacaagatc agcaaattta ttgattggct taaggagaaa atagtaccag cagccaggga 4200caacaagatc agcaaattta ttgattggct taaggagaaa atagtaccag cagccaggga 4200

gaaggttgaa ttcctaaata acttgaaaca gctgccactg ctagagaatc agatctcgaa 4260gaaggttgaa ttcctaaata acttgaaaca gctgccactg ctagagaatc agatctcgaa 4260

cttggaacaa tctgctgctt cacaagagga ccttgaagtc atgtttggga atgtgtcgta 4320cttggaacaa tctgctgctt cacaagagga ccttgaagtc atgtttggga atgtgtcgta 4320

cctagctcac ttctgtcgca agtttcaacc gctatacgcc acggaagcta aaagagtcta 4380cctagctcac ttctgtcgca agtttcaacc gctatacgcc acggaagcta aaagagtcta 4380

tgccctggag aagagaatga ataactatat gcagttcaag agcaaacacc gaattgaacc 4440tgccctggag aagagaatga ataactatat gcagttcaag agcaaacacc gaattgaacc 4440

tgtatgtctc attattaggg gctcaccagg caccgggaag tctctagcca ctggtattat 4500tgtatgtctc attattaggg gctcaccagg caccgggaag tctctagcca ctggtattat 4500

tgctcgagca atcgctgata agtaccactc cagcgtgtac tcgctcccac cagacccgga 4560tgctcgagca atcgctgata agtaccactc cagcgtgtac tcgctcccac cagacccgga 4560

tcattttgac ggttacaagc aacaggtggt tacagtgatg gatgatttgt gtcaaaaccc 4620tcattttgac ggttacaagc aacaggtggt tacagtgatg gatgatttgt gtcaaaaccc 4620

cgatggtaag gatatgtcct tattctgtca aatggtatcc accgtagatt tcattccacc 4680cgatggtaag gatatgtcct tattctgtca aatggtatcc accgtagatt tcattccacc 4680

aatggcttct ctcgaggaga agggagtttc cttcacctct aagtttgtca tcgcatccac 4740aatggcttct ctcgaggaga agggagtttc cttcacctct aagtttgtca tcgcatccac 4740

taatgccagt aatatcatag taccaacagt gtctgattct gacgctattc gccgcaggtt 4800taatgccagt aatatcatag taccaacagt gtctgattct gacgctattc gccgcaggtt 4800

ctacatggac tgtgacattg aagtgacaga ctcgtacaaa acagatctag gtagactgga 4860ctacatggac tgtgacattg aagtgacaga ctcgtacaaa acagatctag gtagactgga 4860

tgcagggcga gccgctaaac tgtgttctga aaataacact gcaaatttca aacgttgcag 4920tgcagggcga gccgctaaac tgtgttctga aaataacact gcaaatttca aacgttgcag 4920

cccattagtg tgtgggaaag ccatccaact tagagataga aagtctaaag tcagatacag 4980cccattagtg tgtgggaaag ccatccaact tagagataga aagtctaaag tcagatacag 4980

tgtggatacg gtggtttcag aacttattag ggaatacagc aataggtccg ccattggtaa 5040tgtggatacg gtggtttcag aacttattag ggaatacagc aataggtccg ccattggtaa 5040

cacaatcgag gctcttttcc aaggtccacc caagttcagg ccaattagga ttagccttga 5100cacaatcgag gctcttttcc aaggtccacc caagttcagg ccaattagga ttagccttga 5100

agaaaaacca gccccagacg ctattagcga tctccttgct agtgtagata gtgaagaagt 5160agaaaaacca gccccagacg ctattagcga tctccttgct agtgtagata gtgaagaagt 5160

gcgccagtac tgcagggatc aaggctggat tattcctgaa gctcccacca atgtggagcg 5220gcgccagtac tgcagggatc aaggctggat tattcctgaa gctcccacca atgtggagcg 5220

gcaccttaat agagcggtgc tcgtcatgca atccatcacc acagtagtgg cggttgtttc 5280gcaccttaat agagcggtgc tcgtcatgca atccatcacc acagtagtgg cggttgtttc 5280

gttggtgtac gtcatctaca agctctttgc agggtttcag ggtgcatatt ctggtgctcc 5340gttggtgtac gtcatctaca agctctttgc agggtttcag ggtgcatatt ctggtgctcc 5340

taagcaagtg cttaagaaac ctgctcttcg cacagcaaca gtgcagggtc cgagccttga 5400taagcaagtg cttaagaaac ctgctcttcg cacagcaaca gtgcagggtc cgagccttga 5400

ctttgctctc tccctactga gaaggaacat caggcaggtc caaacagacc aagggcattt 5460ctttgctctc tccctactga gaaggaacat caggcaggtc caaacagacc aagggcattt 5460

caccatgttg ggtgttaggg atcgcttagc agtcctccca cgccactcac aacctggcaa 5520caccatgttg ggtgttaggg atcgcttagc agtcctccca cgccactcac aacctggcaa 5520

aaccatttgg attgagcaca aactcgtgaa cgtccttgat gcagttgaac tggtggatga 5580aaccatttgg attgagcaca aactcgtgaa cgtccttgat gcagttgaac tggtggatga 5580

gcaaggagtc aacctggaat taaccctcat cactcttgac accaacgaga agtttaggga 5640gcaaggagtc aacctggaat taaccctcat cactcttgac accaacgaga agtttaggga 5640

tatcaccaaa ttcatcccag aaaatatcag cactgctagc gatgccaccc tagtgatcaa 5700tatcaccaaa ttcatcccag aaaatatcag cactgctagc gatgccaccc tagtgatcaa 5700

cacggagcac atgccgtcaa tgtttgtccc ggtgggtgac gttgtgcagt atggcttttt 5760cacggagcac atgccgtcaa tgtttgtccc ggtgggtgac gttgtgcagt atggcttttt 5760

gaatctcagt ggcaagccta cccatcgcac catgatgtac aattttccta ctaaagcagg 5820gaatctcagt ggcaagccta cccatcgcac catgatgtac aattttccta ctaaagcagg 5820

acagtgtgga ggagtggtga catctgttgg gaaggttgtc ggtattcaca ttggtggcaa 5880acagtgtgga ggagtggtga catctgttgg gaaggttgtc ggtattcaca ttggtggcaa 5880

tggcagacaa ggtttttgcg caggcctcaa aaggagttac tttgctagtg aacaaggaga 5940tggcagacaa ggttttttgcg caggcctcaa aaggagttac tttgctagtg aacaaggaga 5940

gatccagtgg gttaagccca ataaagaaac tggaagactc aacatcaatg gaccaacccg 6000gatccagtgg gttaagccca ataaagaaac tggaagactc aacatcaatg gaccaacccg 6000

caccaagtta gaacctagtg tattccatga catcttcgag ggaaataagg aaccagctgt 6060caccaagtta gaacctagtg tattccatga catcttcgag ggaaataagg aaccagctgt 6060

cttgcacagt aaagaccccc gacttgaggt agattttgaa caggccctgt tctctaagta 6120cttgcacagt aaagaccccc gacttgaggt agattttgaa caggccctgt tctctaagta 6120

tgtgggaaac acactacatg agcctgacga gtacatcaaa gaggcagctc tacattatgc 6180tgtgggaaac acactacatg agcctgacga gtacatcaaa gaggcagctc tacattatgc 6180

aaaccaatta aagcaactag aaatcaatac ctctcaaatg agcatggagg aggcctgcta 6240aaaccaatta aagcaactag aaatcaatac ctctcaaatg agcatggagg aggcctgcta 6240

tggtactgag aatcttgagg ctattgatct tcacactagt gcaggttacc cctatagtgc 6300tggtactgag aatcttgagg ctattgatct tcacactagt gcaggttacc cctatagtgc 6300

cctagggata aagaaaagag acatcttaga ccctaccacc agggacgtga gtagaatgaa 6360cctagggata aagaaaagag acatcttaga ccctaccacc agggacgtga gtagaatgaa 6360

gttctacatg gacaagtatg gtcttgatct tccctactcc acttatgtca aggacgagct 6420gttctacatg gacaagtatg gtcttgatct tccctactcc acttatgtca aggacgagct 6420

acgctcgatt gataaaatca agaaagggaa gtcccgcctg atcgaggcca gtagtctaaa 6480acgctcgatt gataaaatca agaaagggaa gtcccgcctg atcgaggcca gtagtctaaa 6480

tgattcagtg tacctcagaa tggctttcgg gcatttgtat gaggctttcc acgcaaatcc 6540tgattcagtg tacctcagaa tggctttcgg gcatttgtat gaggctttcc acgcaaatcc 6540

tgggacgata actggatcgg ccgtggggtg taaccctgac acattctgga gcaagctgcc 6600tgggacgata actggatcgg ccgtggggtg taaccctgac acattctgga gcaagctgcc 6600

aattttgctc cctggttcac tctttgcctt tgactactca ggctatgatg ccagccttag 6660aattttgctc cctggttcac tctttgcctt tgactactca ggctatgatg ccagccttag 6660

ccctgtctgg ttcagagcat tagaattggt tcttagggag atagggtata gtgaagaggc 6720ccctgtctgg ttcagagcat tagaattggt tcttagggag atagggtata gtgaagaggc 6720

aatctcactc attgagggaa tcaaccacac acatcatgtg tatcgtaata agacctattg 6780aatctcactc attgagggaa tcaaccacac acatcatgtg tatcgtaata agacctattg 6780

cgtgcttggt gggatgccct caggctgttc aggaacatcc atcttcaact caatgatcaa 6840cgtgcttggt gggatgccct caggctgttc aggaacatcc atcttcaact caatgatcaa 6840

caacattatt atcagagcac tgctcataaa aacatttaag ggcattgatt tggatgaact 6900caacattatt atcagagcac tgctcataaa aacatttaag ggcattgatt tggatgaact 6900

caacatggtc gcttatggag acgatgtgct cgctagctat cccttcccaa ttgattgctt 6960caacatggtc gcttatggag acgatgtgct cgctagctat cccttcccaa ttgattgctt 6960

ggaactagca aagactggta aggagtatgg tctgaccatg acccctgctg ataaatctcc 7020ggaactagca aagactggta aggagtatgg tctgaccatg acccctgctg ataaatctcc 7020

ttgctttaat gaggtcaatt ggggtaatgc gaccttcctc aaaaggggct ttttgcccga 7080ttgctttaat gaggtcaatt ggggtaatgc gaccttcctc aaaaggggct ttttgcccga 7080

tgaacagttt ccatttttga ttcaccctac tatgccaatg agggagatcc atgagtccat 7140tgaacagttt ccatttttga ttcaccctac tatgccaatg agggagatcc atgagtccat 7140

tcgatggacc aaggacgcac ggaacactca agatcatgtg cggtccttgt gcctcctagc 7200tcgatggacc aaggacgcac ggaacactca agatcatgtg cggtccttgt gcctcctagc 7200

atggcataat ggtaagcaag aatacgagaa gtttgtgagc acaattaggt ctgtcccagt 7260atggcataat ggtaagcaag aatacgagaa gtttgtgagc acaattaggt ctgtcccagt 7260

agggagagcg ttggctattc caaattatga aaatcttaga cgaaattggc tcgagttatt 7320agggagagcg ttggctattc caaattatga aaatcttaga cgaaattggc tcgagttatt 7320

ttagaggtta tacacacctc aaccccacca gaaatctggt cgtgaatgtg actggtgggg 7380ttagaggtta tacacacctc aaccccacca gaaatctggt cgtgaatgtg actggtgggg 7380

gtaaatttgt tataaccaga atagc 7405gtaaatttgt tataaccaga atagc 7405

<210> 3<210> 3

<211> 1987<211> 1987

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 3<400> 3

agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt 60agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt 60

gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg 120gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg 120

atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg 180atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg 180

gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg 240gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg 240

gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc 300gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc 300

acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg 360acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg 360

cgtttcccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt 420cgtttcccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt 420

cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg 480cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg 480

cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct 540cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct 540

tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag 600tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag 600

cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa 660cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa 660

actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag 720actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag 720

ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag 780ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag 780

caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa ggggtctgac 840caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa ggggtctgac 840

gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 900gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 900

ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 960ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 960

taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 1020taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 1020

ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 1080ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 1080

ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 1140ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 1140

gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 1200gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 1200

ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 1260ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 1260

gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg 1320gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg 1320

tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 1380tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 1380

atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 1440atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 1440

gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 1500gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 1500

tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 1560tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 1560

atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc 1620atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc 1620

agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 1680agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 1680

ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 1740ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 1740

tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 1800tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 1800

aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 1860aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 1860

tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 1920tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 1920

aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtgtcgacg 1980aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtgtcgacg 1980

cggccgc 1987cggccgc 1987

<210> 4<210> 4

<211> 2193<211> 2193

<212> PRT<212> PRT

<213> Artificial<213> Artificial

<400> 4<400> 4

Met Gly Ser Gln Val Ser Thr Gln Arg Ser Gly Ser Tyr Glu Asn SerMet Gly Ser Gln Val Ser Thr Gln Arg Ser Gly Ser Tyr Glu Asn Ser

1 5 10 151 5 10 15

Asn Ser Ala Thr Glu Gly Ser Thr Ile Asn Tyr Thr Thr Ile Asn TyrAsn Ser Ala Thr Glu Gly Ser Thr Ile Asn Tyr Thr Thr Ile Asn Tyr

20 25 30 20 25 30

Tyr Lys Asp Ser Tyr Ala Ala Thr Ala Gly Lys Gln Ser Leu Lys GlnTyr Lys Asp Ser Tyr Ala Ala Thr Ala Gly Lys Gln Ser Leu Lys Gln

35 40 45 35 40 45

Asp Pro Asp Lys Phe Ala Asn Pro Val Lys Asp Ile Phe Thr Glu MetAsp Pro Asp Lys Phe Ala Asn Pro Val Lys Asp Ile Phe Thr Glu Met

50 55 60 50 55 60

Ala Ala Pro Leu Lys Ser Pro Ser Ala Glu Ala Cys Gly Tyr Ser AspAla Ala Pro Leu Lys Ser Pro Ser Ala Glu Ala Cys Gly Tyr Ser Asp

65 70 75 8065 70 75 80

Arg Val Ala Gln Leu Thr Ile Gly Asn Ser Thr Ile Thr Thr Gln GluArg Val Ala Gln Leu Thr Ile Gly Asn Ser Thr Ile Thr Thr Gln Glu

85 90 95 85 90 95

Ala Ala Asn Ile Ile Val Gly Tyr Gly Glu Trp Pro Ser Tyr Cys SerAla Ala Asn Ile Ile Val Gly Tyr Gly Glu Trp Pro Ser Tyr Cys Ser

100 105 110 100 105 110

Asp Ser Asp Ala Thr Ala Val Asp Lys Pro Thr Arg Pro Asp Val SerAsp Ser Asp Ala Thr Ala Val Asp Lys Pro Thr Arg Pro Asp Val Ser

115 120 125 115 120 125

Val Asn Arg Phe Tyr Thr Leu Asp Thr Lys Leu Trp Glu Lys Ser SerVal Asn Arg Phe Tyr Thr Leu Asp Thr Lys Leu Trp Glu Lys Ser Ser

130 135 140 130 135 140

Lys Gly Trp Tyr Trp Lys Phe Pro Asp Val Leu Thr Glu Thr Gly ValLys Gly Trp Tyr Trp Lys Phe Pro Asp Val Leu Thr Glu Thr Gly Val

145 150 155 160145 150 155 160

Phe Gly Gln Asn Ala Gln Phe His Tyr Leu Tyr Arg Ser Gly Phe CysPhe Gly Gln Asn Ala Gln Phe His Tyr Leu Tyr Arg Ser Gly Phe Cys

165 170 175 165 170 175

Ile His Val Gln Cys Asn Ala Ser Lys Phe His Gln Gly Ala Leu LeuIle His Val Gln Cys Asn Ala Ser Lys Phe His Gln Gly Ala Leu Leu

180 185 190 180 185 190

Val Ala Val Leu Pro Glu Tyr Val Ile Gly Thr Val Ala Gly Gly ThrVal Ala Val Leu Pro Glu Tyr Val Ile Gly Thr Val Ala Gly Gly Thr

195 200 205 195 200 205

Gly Thr Glu Asp Thr His Pro Pro Tyr Lys Gln Thr Gln Pro Gly AlaGly Thr Glu Asp Thr His Pro Pro Tyr Lys Gln Thr Gln Pro Gly Ala

210 215 220 210 215 220

Asp Gly Phe Glu Leu Gln His Pro Tyr Val Leu Asp Ala Gly Ile ProAsp Gly Phe Glu Leu Gln His Pro Tyr Val Leu Asp Ala Gly Ile Pro

225 230 235 240225 230 235 240

Ile Ser Gln Leu Thr Val Cys Pro His Gln Trp Ile Asn Leu Arg ThrIle Ser Gln Leu Thr Val Cys Pro His Gln Trp Ile Asn Leu Arg Thr

245 250 255 245 250 255

Asn Asn Cys Ala Thr Ile Ile Val Pro Tyr Ile Asn Ala Leu Pro PheAsn Asn Cys Ala Thr Ile Ile Ile Val Pro Tyr Ile Asn Ala Leu Pro Phe

260 265 270 260 265 270

Asp Ser Ala Leu Asn His Cys Asn Phe Gly Leu Leu Val Val Pro IleAsp Ser Ala Leu Asn His Cys Asn Phe Gly Leu Leu Val Val Pro Ile

275 280 285 275 280 285

Ser Pro Leu Asp Tyr Asp Gln Gly Ala Thr Pro Val Ile Pro Ile ThrSer Pro Leu Asp Tyr Asp Gln Gly Ala Thr Pro Val Ile Pro Ile Thr

290 295 300 290 295 300

Ile Thr Leu Ala Pro Met Cys Ser Glu Phe Ala Gly Leu Arg Gln AlaIle Thr Leu Ala Pro Met Cys Ser Glu Phe Ala Gly Leu Arg Gln Ala

305 310 315 320305 310 315 320

Val Thr Gln Gly Phe Pro Thr Glu Leu Lys Pro Gly Thr Asn Gln PheVal Thr Gln Gly Phe Pro Thr Glu Leu Lys Pro Gly Thr Asn Gln Phe

325 330 335 325 330 335

Leu Thr Thr Asp Asp Gly Val Ser Ala Pro Ile Leu Pro Asn Phe HisLeu Thr Thr Asp Asp Gly Val Ser Ala Pro Ile Leu Pro Asn Phe His

340 345 350 340 345 350

Pro Thr Pro Cys Ile His Ile Pro Gly Glu Val Arg Asn Leu Leu GluPro Thr Pro Cys Ile His Ile Pro Gly Glu Val Arg Asn Leu Leu Glu

355 360 365 355 360 365

Leu Cys Gln Val Glu Thr Ile Leu Glu Val Asn Asn Val Pro Thr AsnLeu Cys Gln Val Glu Thr Ile Leu Glu Val Asn Asn Val Pro Thr Asn

370 375 380 370 375 380

Ala Thr Ser Leu Met Glu Arg Leu Arg Phe Pro Val Ser Ala Gln AlaAla Thr Ser Leu Met Glu Arg Leu Arg Phe Pro Val Ser Ala Gln Ala

385 390 395 400385 390 395 400

Gly Lys Gly Glu Leu Cys Ala Val Phe Arg Ala Asp Pro Gly Arg AsnGly Lys Gly Glu Leu Cys Ala Val Phe Arg Ala Asp Pro Gly Arg Asn

405 410 415 405 410 415

Gly Pro Trp Gln Ser Thr Leu Leu Gly Gln Leu Cys Gly Tyr Tyr ThrGly Pro Trp Gln Ser Thr Leu Leu Gly Gln Leu Cys Gly Tyr Tyr Thr

420 425 430 420 425 430

Gln Trp Ser Gly Ser Leu Glu Val Thr Phe Met Phe Thr Gly Ser PheGln Trp Ser Gly Ser Leu Glu Val Thr Phe Met Phe Thr Gly Ser Phe

435 440 445 435 440 445

Met Ala Thr Gly Lys Met Leu Ile Ala Tyr Thr Pro Pro Gly Gly ProMet Ala Thr Gly Lys Met Leu Ile Ala Tyr Thr Pro Pro Gly Gly Pro

450 455 460 450 455 460

Leu Pro Lys Asp Arg Ala Thr Ala Met Leu Gly Thr His Val Ile TrpLeu Pro Lys Asp Arg Ala Thr Ala Met Leu Gly Thr His Val Ile Trp

465 470 475 480465 470 475 480

Asp Phe Gly Leu Gln Ser Ser Val Thr Leu Val Ile Pro Trp Ile SerAsp Phe Gly Leu Gln Ser Ser Val Thr Leu Val Ile Pro Trp Ile Ser

485 490 495 485 490 495

Asn Thr His Tyr Arg Ala His Ala Arg Asp Gly Val Phe Asp Tyr TyrAsn Thr His Tyr Arg Ala His Ala Arg Asp Gly Val Phe Asp Tyr Tyr

500 505 510 500 505 510

Thr Thr Gly Leu Val Ser Ile Trp Tyr Gln Thr Asn Tyr Val Val ProThr Thr Gly Leu Val Ser Ile Trp Tyr Gln Thr Asn Tyr Val Val Pro

515 520 525 515 520 525

Ile Gly Ala Pro Asn Thr Ala Tyr Ile Ile Ala Leu Ala Ala Ala GlnIle Gly Ala Pro Asn Thr Ala Tyr Ile Ile Ala Leu Ala Ala Ala Gln

530 535 540 530 535 540

Lys Asn Phe Thr Met Lys Leu Cys Lys Asp Ala Ser Asp Ile Leu GlnLys Asn Phe Thr Met Lys Leu Cys Lys Asp Ala Ser Asp Ile Leu Gln

545 550 555 560545 550 555 560

Thr Gly Thr Ile Gln Gly Asp Arg Val Ala Asp Val Ile Glu Ser SerThr Gly Thr Ile Gln Gly Asp Arg Val Ala Asp Val Ile Glu Ser Ser

565 570 575 565 570 575

Ile Gly Asp Ser Val Ser Arg Ala Leu Thr His Ala Leu Pro Ala ProIle Gly Asp Ser Val Ser Arg Ala Leu Thr His Ala Leu Pro Ala Pro

580 585 590 580 585 590

Thr Gly Gln Asn Thr Gln Val Ser Ser His Arg Leu Asp Thr Gly LysThr Gly Gln Asn Thr Gln Val Ser Ser His Arg Leu Asp Thr Gly Lys

595 600 605 595 600 605

Val Pro Ala Leu Gln Ala Ala Glu Ile Gly Ala Ser Ser Asn Ala SerVal Pro Ala Leu Gln Ala Ala Glu Ile Gly Ala Ser Ser Asn Ala Ser

610 615 620 610 615 620

Asp Glu Ser Met Ile Glu Thr Arg Cys Val Leu Asn Ser His Ser ThrAsp Glu Ser Met Ile Glu Thr Arg Cys Val Leu Asn Ser His Ser Thr

625 630 635 640625 630 635 640

Ala Glu Thr Thr Leu Asp Ser Phe Phe Ser Arg Ala Gly Leu Val GlyAla Glu Thr Thr Leu Asp Ser Phe Phe Ser Arg Ala Gly Leu Val Gly

645 650 655 645 650 655

Glu Ile Asp Leu Pro Leu Glu Gly Thr Thr Asn Pro Asn Gly Tyr AlaGlu Ile Asp Leu Pro Leu Glu Gly Thr Thr Asn Pro Asn Gly Tyr Ala

660 665 670 660 665 670

Asn Trp Asp Ile Asp Ile Thr Gly Tyr Ala Gln Met Arg Arg Lys ValAsn Trp Asp Ile Asp Ile Thr Gly Tyr Ala Gln Met Arg Arg Lys Val

675 680 685 675 680 685

Glu Leu Phe Thr Tyr Met Arg Phe Asp Ala Glu Phe Thr Phe Val AlaGlu Leu Phe Thr Tyr Met Arg Phe Asp Ala Glu Phe Thr Phe Val Ala

690 695 700 690 695 700

Cys Thr Pro Thr Gly Glu Val Val Pro Gln Leu Leu Gln Tyr Met PheCys Thr Pro Thr Gly Glu Val Val Pro Gln Leu Leu Gln Tyr Met Phe

705 710 715 720705 710 715 720

Val Pro Pro Gly Ala Pro Lys Pro Asp Ser Arg Glu Ser Leu Ala TrpVal Pro Pro Gly Ala Pro Lys Pro Asp Ser Arg Glu Ser Leu Ala Trp

725 730 735 725 730 735

Gln Thr Ala Thr Asn Pro Ser Val Phe Val Lys Leu Ser Asp Pro ProGln Thr Ala Thr Asn Pro Ser Val Phe Val Lys Leu Ser Asp Pro Pro

740 745 750 740 745 750

Ala Gln Val Ser Val Pro Phe Met Ser Pro Ala Ser Ala Tyr Gln TrpAla Gln Val Ser Val Pro Phe Met Ser Pro Ala Ser Ala Tyr Gln Trp

755 760 765 755 760 765

Phe Tyr Asp Gly Tyr Pro Thr Phe Gly Glu His Lys Gln Glu Lys AspPhe Tyr Asp Gly Tyr Pro Thr Phe Gly Glu His Lys Gln Glu Lys Asp

770 775 780 770 775 780

Leu Glu Tyr Gly Ala Cys Pro Asn Asn Met Met Gly Thr Phe Ser ValLeu Glu Tyr Gly Ala Cys Pro Asn Asn Met Met Gly Thr Phe Ser Val

785 790 795 800785 790 795 800

Arg Thr Val Gly Thr Ser Lys Ser Lys Tyr Pro Leu Val Val Arg IleArg Thr Val Gly Thr Ser Lys Ser Lys Tyr Pro Leu Val Val Arg Ile

805 810 815 805 810 815

Tyr Met Arg Met Lys His Val Arg Ala Trp Ile Pro Arg Pro Met ArgTyr Met Arg Met Lys His Val Arg Ala Trp Ile Pro Arg Pro Met Arg

820 825 830 820 825 830

Asn Gln Asn Tyr Leu Phe Lys Ala Asn Pro Asn Tyr Ala Gly Asn SerAsn Gln Asn Tyr Leu Phe Lys Ala Asn Pro Asn Tyr Ala Gly Asn Ser

835 840 845 835 840 845

Ile Lys Pro Thr Gly Ala Ser Arg Thr Ala Ile Thr Thr Leu Gly LysIle Lys Pro Thr Gly Ala Ser Arg Thr Ala Ile Thr Thr Leu Gly Lys

850 855 860 850 855 860

Phe Gly Gln Gln Ser Gly Ala Ile Tyr Val Gly Asn Phe Arg Val ValPhe Gly Gln Gln Ser Gly Ala Ile Tyr Val Gly Asn Phe Arg Val Val

865 870 875 880865 870 875 880

Asn Arg His Leu Ala Thr His Asn Asp Trp Ala Asn Leu Val Trp GluAsn Arg His Leu Ala Thr His Asn Asp Trp Ala Asn Leu Val Trp Glu

885 890 895 885 890 895

Asp Ser Ser Arg Asp Leu Leu Val Ser Ser Thr Thr Ala Gln Gly CysAsp Ser Ser Arg Asp Leu Leu Val Ser Ser Thr Thr Ala Gln Gly Cys

900 905 910 900 905 910

Asp Thr Ile Ala Arg Cys Asp Cys Gln Thr Gly Val Tyr Tyr Cys AsnAsp Thr Ile Ala Arg Cys Asp Cys Gln Thr Gly Val Tyr Tyr Cys Asn

915 920 925 915 920 925

Ser Met Arg Lys His Tyr Pro Val Ser Phe Ser Lys Pro Ser Leu IleSer Met Arg Lys His Tyr Pro Val Ser Phe Ser Lys Pro Ser Leu Ile

930 935 940 930 935 940

Tyr Val Glu Ala Ser Glu Tyr Tyr Pro Ala Arg Tyr Gln Ser His LeuTyr Val Glu Ala Ser Glu Tyr Tyr Pro Ala Arg Tyr Gln Ser His Leu

945 950 955 960945 950 955 960

Met Leu Ala Gln Gly His Ser Glu Pro Gly Asp Cys Gly Gly Ile LeuMet Leu Ala Gln Gly His Ser Glu Pro Gly Asp Cys Gly Gly Ile Leu

965 970 975 965 970 975

Arg Cys Gln His Gly Val Ile Gly Ile Val Ser Thr Gly Gly Asn GlyArg Cys Gln His Gly Val Ile Gly Ile Val Ser Thr Gly Gly Asn Gly

980 985 990 980 985 990

Leu Val Gly Phe Ala Asp Val Arg Asp Leu Leu Trp Leu Asp Glu GluLeu Val Gly Phe Ala Asp Val Arg Asp Leu Leu Trp Leu Asp Glu Glu

995 1000 1005 995 1000 1005

Ala Met Glu Gln Gly Val Ser Asp Tyr Ile Lys Gly Leu Gly Asp AlaAla Met Glu Gln Gly Val Ser Asp Tyr Ile Lys Gly Leu Gly Asp Ala

1010 1015 1020 1010 1015 1020

Phe Gly Thr Gly Phe Thr Asp Ala Val Ser Arg Glu Val Glu Ala LeuPhe Gly Thr Gly Phe Thr Asp Ala Val Ser Arg Glu Val Glu Ala Leu

1025 1030 1035 10401025 1030 1035 1040

Lys Asn Tyr Leu Ile Gly Ser Glu Gly Ala Val Glu Lys Ile Leu LysLys Asn Tyr Leu Ile Gly Ser Glu Gly Ala Val Glu Lys Ile Leu Lys

1045 1050 1055 1045 1050 1055

Asn Leu Ile Lys Leu Ile Ser Ala Leu Val Ile Val Ile Arg Ser AspAsn Leu Ile Lys Leu Ile Ser Ala Leu Val Ile Val Ile Arg Ser Asp

1060 1065 1070 1060 1065 1070

Tyr Asp Met Val Thr Leu Thr Ala Thr Leu Ala Leu Ile Gly Cys HisTyr Asp Met Val Thr Leu Thr Ala Thr Leu Ala Leu Ile Gly Cys His

1075 1080 1085 1075 1080 1085

Gly Ser Pro Trp Ala Trp Ile Lys Ala Lys Thr Ala Ser Ile Leu GlyGly Ser Pro Trp Ala Trp Ile Lys Ala Lys Thr Ala Ser Ile Leu Gly

1090 1095 1100 1090 1095 1100

Ile Pro Ile Ala Gln Lys Gln Ser Ala Ser Trp Leu Lys Lys Phe AsnIle Pro Ile Ala Gln Lys Gln Ser Ala Ser Trp Leu Lys Lys Phe Asn

1105 1110 1115 11201105 1110 1115 1120

Asp Met Ala Asn Ala Ala Lys Gly Leu Glu Trp Val Ser Asn Lys IleAsp Met Ala Asn Ala Ala Lys Gly Leu Glu Trp Val Ser Asn Lys Ile

1125 1130 1135 1125 1130 1135

Ser Lys Phe Ile Asp Trp Leu Lys Glu Lys Ile Val Pro Ala Ala ArgSer Lys Phe Ile Asp Trp Leu Lys Glu Lys Ile Val Pro Ala Ala Arg

1140 1145 1150 1140 1145 1150

Glu Lys Val Glu Phe Leu Asn Asn Leu Lys Gln Leu Pro Leu Leu GluGlu Lys Val Glu Phe Leu Asn Asn Leu Lys Gln Leu Pro Leu Leu Glu

1155 1160 1165 1155 1160 1165

Asn Gln Ile Ser Asn Leu Glu Gln Ser Ala Ala Ser Gln Glu Asp LeuAsn Gln Ile Ser Asn Leu Glu Gln Ser Ala Ala Ser Gln Glu Asp Leu

1170 1175 1180 1170 1175 1180

Glu Val Met Phe Gly Asn Val Ser Tyr Leu Ala His Phe Cys Arg LysGlu Val Met Phe Gly Asn Val Ser Tyr Leu Ala His Phe Cys Arg Lys

1185 1190 1195 12001185 1190 1195 1200

Phe Gln Pro Leu Tyr Ala Thr Glu Ala Lys Arg Val Tyr Ala Leu GluPhe Gln Pro Leu Tyr Ala Thr Glu Ala Lys Arg Val Tyr Ala Leu Glu

1205 1210 1215 1205 1210 1215

Lys Arg Met Asn Asn Tyr Met Gln Phe Lys Ser Lys His Arg Ile GluLys Arg Met Asn Asn Tyr Met Gln Phe Lys Ser Lys His Arg Ile Glu

1220 1225 1230 1220 1225 1230

Pro Val Cys Leu Ile Ile Arg Gly Ser Pro Gly Thr Gly Lys Ser LeuPro Val Cys Leu Ile Ile Arg Gly Ser Pro Gly Thr Gly Lys Ser Leu

1235 1240 1245 1235 1240 1245

Ala Thr Gly Ile Ile Ala Arg Ala Ile Ala Asp Lys Tyr His Ser SerAla Thr Gly Ile Ile Ala Arg Ala Ile Ala Asp Lys Tyr His Ser Ser

1250 1255 1260 1250 1255 1260

Val Tyr Ser Leu Pro Pro Asp Pro Asp His Phe Asp Gly Tyr Lys GlnVal Tyr Ser Leu Pro Pro Asp Pro Asp His Phe Asp Gly Tyr Lys Gln

1265 1270 1275 12801265 1270 1275 1280

Gln Val Val Thr Val Met Asp Asp Leu Cys Gln Asn Pro Asp Gly LysGln Val Val Thr Val Met Asp Asp Asp Leu Cys Gln Asn Pro Asp Gly Lys

1285 1290 1295 1285 1290 1295

Asp Met Ser Leu Phe Cys Gln Met Val Ser Thr Val Asp Phe Ile ProAsp Met Ser Leu Phe Cys Gln Met Val Ser Thr Val Asp Phe Ile Pro

1300 1305 1310 1300 1305 1310

Pro Met Ala Ser Leu Glu Glu Lys Gly Val Ser Phe Thr Ser Lys PhePro Met Ala Ser Leu Glu Glu Lys Gly Val Ser Phe Thr Ser Lys Phe

1315 1320 1325 1315 1320 1325

Val Ile Ala Ser Thr Asn Ala Ser Asn Ile Ile Val Pro Thr Val SerVal Ile Ala Ser Thr Asn Ala Ser Asn Ile Ile Val Pro Thr Val Ser

1330 1335 1340 1330 1335 1340

Asp Ser Asp Ala Ile Arg Arg Arg Phe Tyr Met Asp Cys Asp Ile GluAsp Ser Asp Ala Ile Arg Arg Arg Phe Tyr Met Asp Cys Asp Ile Glu

1345 1350 1355 13601345 1350 1355 1360

Val Thr Asp Ser Tyr Lys Thr Asp Leu Gly Arg Leu Asp Ala Gly ArgVal Thr Asp Ser Tyr Lys Thr Asp Leu Gly Arg Leu Asp Ala Gly Arg

1365 1370 1375 1365 1370 1375

Ala Ala Lys Leu Cys Ser Glu Asn Asn Thr Ala Asn Phe Lys Arg CysAla Ala Lys Leu Cys Ser Glu Asn Asn Thr Ala Asn Phe Lys Arg Cys

1380 1385 1390 1380 1385 1390

Ser Pro Leu Val Cys Gly Lys Ala Ile Gln Leu Arg Asp Arg Lys SerSer Pro Leu Val Cys Gly Lys Ala Ile Gln Leu Arg Asp Arg Lys Ser

1395 1400 1405 1395 1400 1405

Lys Val Arg Tyr Ser Val Asp Thr Val Val Ser Glu Leu Ile Arg GluLys Val Arg Tyr Ser Val Asp Thr Val Val Ser Glu Leu Ile Arg Glu

1410 1415 1420 1410 1415 1420

Tyr Ser Asn Arg Ser Ala Ile Gly Asn Thr Ile Glu Ala Leu Phe GlnTyr Ser Asn Arg Ser Ala Ile Gly Asn Thr Ile Glu Ala Leu Phe Gln

1425 1430 1435 14401425 1430 1435 1440

Gly Pro Pro Lys Phe Arg Pro Ile Arg Ile Ser Leu Glu Glu Lys ProGly Pro Pro Lys Phe Arg Pro Ile Arg Ile Ser Leu Glu Glu Lys Pro

1445 1450 1455 1445 1450 1455

Ala Pro Asp Ala Ile Ser Asp Leu Leu Ala Ser Val Asp Ser Glu GluAla Pro Asp Ala Ile Ser Asp Leu Leu Ala Ser Val Asp Ser Glu Glu

1460 1465 1470 1460 1465 1470

Val Arg Gln Tyr Cys Arg Asp Gln Gly Trp Ile Ile Pro Glu Ala ProVal Arg Gln Tyr Cys Arg Asp Gln Gly Trp Ile Ile Pro Glu Ala Pro

1475 1480 1485 1475 1480 1485

Thr Asn Val Glu Arg His Leu Asn Arg Ala Val Leu Val Met Gln SerThr Asn Val Glu Arg His Leu Asn Arg Ala Val Leu Val Met Gln Ser

1490 1495 1500 1490 1495 1500

Ile Thr Thr Val Val Ala Val Val Ser Leu Val Tyr Val Ile Tyr LysIle Thr Thr Val Val Ala Val Val Ser Leu Val Tyr Val Ile Tyr Lys

1505 1510 1515 15201505 1510 1515 1520

Leu Phe Ala Gly Phe Gln Gly Ala Tyr Ser Gly Ala Pro Lys Gln ValLeu Phe Ala Gly Phe Gln Gly Ala Tyr Ser Gly Ala Pro Lys Gln Val

1525 1530 1535 1525 1530 1535

Leu Lys Lys Pro Ala Leu Arg Thr Ala Thr Val Gln Gly Pro Ser LeuLeu Lys Lys Pro Ala Leu Arg Thr Ala Thr Val Gln Gly Pro Ser Leu

1540 1545 1550 1540 1545 1550

Asp Phe Ala Leu Ser Leu Leu Arg Arg Asn Ile Arg Gln Val Gln ThrAsp Phe Ala Leu Ser Leu Leu Arg Arg Asn Ile Arg Gln Val Gln Thr

1555 1560 1565 1555 1560 1565

Asp Gln Gly His Phe Thr Met Leu Gly Val Arg Asp Arg Leu Ala ValAsp Gln Gly His Phe Thr Met Leu Gly Val Arg Asp Arg Leu Ala Val

1570 1575 1580 1570 1575 1580

Leu Pro Arg His Ser Gln Pro Gly Lys Thr Ile Trp Ile Glu His LysLeu Pro Arg His Ser Gln Pro Gly Lys Thr Ile Trp Ile Glu His Lys

1585 1590 1595 16001585 1590 1595 1600

Leu Val Asn Val Leu Asp Ala Val Glu Leu Val Asp Glu Gln Gly ValLeu Val Asn Val Leu Asp Ala Val Glu Leu Val Asp Glu Gln Gly Val

1605 1610 1615 1605 1610 1615

Asn Leu Glu Leu Thr Leu Ile Thr Leu Asp Thr Asn Glu Lys Phe ArgAsn Leu Glu Leu Thr Leu Ile Thr Leu Asp Thr Asn Glu Lys Phe Arg

1620 1625 1630 1620 1625 1630

Asp Ile Thr Lys Phe Ile Pro Glu Asn Ile Ser Thr Ala Ser Asp AlaAsp Ile Thr Lys Phe Ile Pro Glu Asn Ile Ser Thr Ala Ser Asp Ala

1635 1640 1645 1635 1640 1645

Thr Leu Val Ile Asn Thr Glu His Met Pro Ser Met Phe Val Pro ValThr Leu Val Ile Asn Thr Glu His Met Pro Ser Met Phe Val Pro Val

1650 1655 1660 1650 1655 1660

Gly Asp Val Val Gln Tyr Gly Phe Leu Asn Leu Ser Gly Lys Pro ThrGly Asp Val Val Gln Tyr Gly Phe Leu Asn Leu Ser Gly Lys Pro Thr

1665 1670 1675 16801665 1670 1675 1680

His Arg Thr Met Met Tyr Asn Phe Pro Thr Lys Ala Gly Gln Cys GlyHis Arg Thr Met Met Tyr Asn Phe Pro Thr Lys Ala Gly Gln Cys Gly

1685 1690 1695 1685 1690 1695

Gly Val Val Thr Ser Val Gly Lys Val Val Gly Ile His Ile Gly GlyGly Val Val Thr Ser Val Gly Lys Val Val Gly Ile His Ile Gly Gly

1700 1705 1710 1700 1705 1710

Asn Gly Arg Gln Gly Phe Cys Ala Gly Leu Lys Arg Ser Tyr Phe AlaAsn Gly Arg Gln Gly Phe Cys Ala Gly Leu Lys Arg Ser Tyr Phe Ala

1715 1720 1725 1715 1720 1725

Ser Glu Gln Gly Glu Ile Gln Trp Val Lys Pro Asn Lys Glu Thr GlySer Glu Gln Gly Glu Ile Gln Trp Val Lys Pro Asn Lys Glu Thr Gly

1730 1735 1740 1730 1735 1740

Arg Leu Asn Ile Asn Gly Pro Thr Arg Thr Lys Leu Glu Pro Ser ValArg Leu Asn Ile Asn Gly Pro Thr Arg Thr Lys Leu Glu Pro Ser Val

1745 1750 1755 17601745 1750 1755 1760

Phe His Asp Ile Phe Glu Gly Asn Lys Glu Pro Ala Val Leu His SerPhe His Asp Ile Phe Glu Gly Asn Lys Glu Pro Ala Val Leu His Ser

1765 1770 1775 1765 1770 1775

Lys Asp Pro Arg Leu Glu Val Asp Phe Glu Gln Ala Leu Phe Ser LysLys Asp Pro Arg Leu Glu Val Asp Phe Glu Gln Ala Leu Phe Ser Lys

1780 1785 1790 1780 1785 1790

Tyr Val Gly Asn Thr Leu His Glu Pro Asp Glu Tyr Ile Lys Glu AlaTyr Val Gly Asn Thr Leu His Glu Pro Asp Glu Tyr Ile Lys Glu Ala

1795 1800 1805 1795 1800 1805

Ala Leu His Tyr Ala Asn Gln Leu Lys Gln Leu Glu Ile Asn Thr SerAla Leu His Tyr Ala Asn Gln Leu Lys Gln Leu Glu Ile Asn Thr Ser

1810 1815 1820 1810 1815 1820

Gln Met Ser Met Glu Glu Ala Cys Tyr Gly Thr Glu Asn Leu Glu AlaGln Met Ser Met Glu Glu Ala Cys Tyr Gly Thr Glu Asn Leu Glu Ala

1825 1830 1835 18401825 1830 1835 1840

Ile Asp Leu His Thr Ser Ala Gly Tyr Pro Tyr Ser Ala Leu Gly IleIle Asp Leu His Thr Ser Ala Gly Tyr Pro Tyr Ser Ala Leu Gly Ile

1845 1850 1855 1845 1850 1855

Lys Lys Arg Asp Ile Leu Asp Pro Thr Thr Arg Asp Val Ser Arg MetLys Lys Arg Asp Ile Leu Asp Pro Thr Thr Arg Asp Val Ser Arg Met

1860 1865 1870 1860 1865 1870

Lys Phe Tyr Met Asp Lys Tyr Gly Leu Asp Leu Pro Tyr Ser Thr TyrLys Phe Tyr Met Asp Lys Tyr Gly Leu Asp Leu Pro Tyr Ser Thr Tyr

1875 1880 1885 1875 1880 1885

Val Lys Asp Glu Leu Arg Ser Ile Asp Lys Ile Lys Lys Gly Lys SerVal Lys Asp Glu Leu Arg Ser Ile Asp Lys Ile Lys Lys Gly Lys Ser

1890 1895 1900 1890 1895 1900

Arg Leu Ile Glu Ala Ser Ser Leu Asn Asp Ser Val Tyr Leu Arg MetArg Leu Ile Glu Ala Ser Ser Leu Asn Asp Ser Val Tyr Leu Arg Met

1905 1910 1915 19201905 1910 1915 1920

Ala Phe Gly His Leu Tyr Glu Ala Phe His Ala Asn Pro Gly Thr IleAla Phe Gly His Leu Tyr Glu Ala Phe His Ala Asn Pro Gly Thr Ile

1925 1930 1935 1925 1930 1935

Thr Gly Ser Ala Val Gly Cys Asn Pro Asp Thr Phe Trp Ser Lys LeuThr Gly Ser Ala Val Gly Cys Asn Pro Asp Thr Phe Trp Ser Lys Leu

1940 1945 1950 1940 1945 1950

Pro Ile Leu Leu Pro Gly Ser Leu Phe Ala Phe Asp Tyr Ser Gly TyrPro Ile Leu Leu Pro Gly Ser Leu Phe Ala Phe Asp Tyr Ser Gly Tyr

1955 1960 1965 1955 1960 1965

Asp Ala Ser Leu Ser Pro Val Trp Phe Arg Ala Leu Glu Leu Val LeuAsp Ala Ser Leu Ser Pro Val Trp Phe Arg Ala Leu Glu Leu Val Leu

1970 1975 1980 1970 1975 1980

Arg Glu Ile Gly Tyr Ser Glu Glu Ala Ile Ser Leu Ile Glu Gly IleArg Glu Ile Gly Tyr Ser Glu Glu Ala Ile Ser Leu Ile Glu Gly Ile

1985 1990 1995 20001985 1990 1995 2000

Asn His Thr His His Val Tyr Arg Asn Lys Thr Tyr Cys Val Leu GlyAsn His Thr His His Val Tyr Arg Asn Lys Thr Tyr Cys Val Leu Gly

2005 2010 2015 2005 2010 2015

Gly Met Pro Ser Gly Cys Ser Gly Thr Ser Ile Phe Asn Ser Met IleGly Met Pro Ser Gly Cys Ser Gly Thr Ser Ile Phe Asn Ser Met Ile

2020 2025 2030 2020 2025 2030

Asn Asn Ile Ile Ile Arg Ala Leu Leu Ile Lys Thr Phe Lys Gly IleAsn Asn Ile Ile Ile Arg Ala Leu Leu Ile Lys Thr Phe Lys Gly Ile

2035 2040 2045 2035 2040 2045

Asp Leu Asp Glu Leu Asn Met Val Ala Tyr Gly Asp Asp Val Leu AlaAsp Leu Asp Glu Leu Asn Met Val Ala Tyr Gly Asp Asp Val Leu Ala

2050 2055 2060 2050 2055 2060

Ser Tyr Pro Phe Pro Ile Asp Cys Leu Glu Leu Ala Lys Thr Gly LysSer Tyr Pro Phe Pro Ile Asp Cys Leu Glu Leu Ala Lys Thr Gly Lys

2065 2070 2075 20802065 2070 2075 2080

Glu Tyr Gly Leu Thr Met Thr Pro Ala Asp Lys Ser Pro Cys Phe AsnGlu Tyr Gly Leu Thr Met Thr Pro Ala Asp Lys Ser Pro Cys Phe Asn

2085 2090 2095 2085 2090 2095

Glu Val Asn Trp Gly Asn Ala Thr Phe Leu Lys Arg Gly Phe Leu ProGlu Val Asn Trp Gly Asn Ala Thr Phe Leu Lys Arg Gly Phe Leu Pro

2100 2105 2110 2100 2105 2110

Asp Glu Gln Phe Pro Phe Leu Ile His Pro Thr Met Pro Met Arg GluAsp Glu Gln Phe Pro Phe Leu Ile His Pro Thr Met Pro Met Arg Glu

2115 2120 2125 2115 2120 2125

Ile His Glu Ser Ile Arg Trp Thr Lys Asp Ala Arg Asn Thr Gln AspIle His Glu Ser Ile Arg Trp Thr Lys Asp Ala Arg Asn Thr Gln Asp

2130 2135 2140 2130 2135 2140

His Val Arg Ser Leu Cys Leu Leu Ala Trp His Asn Gly Lys Gln GluHis Val Arg Ser Leu Cys Leu Leu Ala Trp His Asn Gly Lys Gln Glu

2145 2150 2155 21602145 2150 2155 2160

Tyr Glu Lys Phe Val Ser Thr Ile Arg Ser Val Pro Val Gly Arg AlaTyr Glu Lys Phe Val Ser Thr Ile Arg Ser Val Pro Val Gly Arg Ala

2165 2170 2175 2165 2170 2175

Leu Ala Ile Pro Asn Tyr Glu Asn Leu Arg Arg Asn Trp Leu Glu LeuLeu Ala Ile Pro Asn Tyr Glu Asn Leu Arg Arg Asn Trp Leu Glu Leu

2180 2185 2190 2180 2185 2190

PhePhe

<210> 5<210> 5

<211> 9982<211> 9982

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 5<400> 5

gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 60gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 60

tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 120tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 120

tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 180tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 180

atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 240atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 240

gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 300gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 300

aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 360aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 360

ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 420ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 420

tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 480tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 480

ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 540ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 540

ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 600ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 600

ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 660ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 660

gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 720gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 720

gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 780gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 780

gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct 840gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct 840

cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 900cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 900

acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 960acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 960

acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1020acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1020

tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1080tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1080

ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1140ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1140

ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 1200ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 1200

tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 1260tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 1260

aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 1320aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 1320

ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 1380ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 1380

ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 1440ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 1440

gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 1500gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 1500

gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 1560gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 1560

cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 1620cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 1620

actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 1680actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 1680

ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 1740ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 1740

tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 1800tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 1800

ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 1860ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 1860

agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 1920agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 1920

aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt gtcgacgcgg 1980aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt gtcgacgcgg 1980

ccgctaatac gactcactat aggttaaaac agcctgtggg ttgcacccac tcacagggcc 2040ccgctaatac gactcactat aggttaaaac agcctgtggg ttgcacccac tcacagggcc 2040

tactgggcgc aagcactctg gtacctcggt acctttgtgc gcctgtttta cacccccccc 2100tactgggcgc aagcactctg gtacctcggt acctttgtgc gcctgtttta cacccccccc 2100

ccaatgaaac ttagaagcaa taaaccacga tcaatagcag gcataacgct ccagttatgt 2160ccaatgaaac ttagaagcaa taaaccacga tcaatagcag gcataacgct ccagttatgt 2160

cttgatcaag cacttctgtt tccccggact gagtatcaat agactgctcg cgcggttgaa 2220cttgatcaag cacttctgtt tccccggact gagtatcaat agactgctcg cgcggttgaa 2220

ggagaaaacg ttcgttatcc ggctaactac ttcggaaaac ctagtaacac catgaaagtt 2280ggagaaaacg ttcgttatcc ggctaactac ttcggaaaac ctagtaacac catgaaagtt 2280

gcggagagct tcgttcagca ctcccccagt gtagatcagg tcgatgagtc accgcgttcc 2340gcggagagct tcgttcagca ctcccccagt gtagatcagg tcgatgagtc accgcgttcc 2340

ccacgggcga ccgtggcggt ggctgcgttg gcggcctgcc catggggtaa cccatggggc 2400ccacgggcga ccgtggcggt ggctgcgttg gcggcctgcc catggggtaa cccatggggc 2400

gctctaatac ggacatggtg tgaagagtct actgagctag ttggtagtcc tccggcccct 2460gctctaatac ggacatggtg tgaagagtct actgagctag ttggtagtcc tccggcccct 2460

gaatgcggct aatcccaact gcggagcaca cgcccacaag ccagcgggta gtgtgtcgta 2520gaatgcggct aatcccaact gcggagcaca cgcccacaag ccagcgggta gtgtgtcgta 2520

acgggtaact ctgcagcgga accgactact ttgggtgtcc gtgtttcctt ttatctttat 2580acgggtaact ctgcagcgga accgactact ttgggtgtcc gtgtttcctt ttatctttat 2580

attggctgct tatggtgaca attaaagaat tgttaccata tagctattgg attagccatc 2640attggctgct tatggtgaca attaaagaat tgttaccata tagctattgg attagccatc 2640

cggtgtgcaa cagagcaatt atttacctat ttattggttt tgtaccatta acctcgaatt 2700cggtgtgcaa cagagcaatt atttacctat ttattggttt tgtaccatta acctcgaatt 2700

ctgtgaccac ccttaattat atcttgaccc ttaacacagc taaactctag aatggtcttc 2760ctgtgaccac ccttaattat atcttgaccc ttaacacagc taaactctag aatggtcttc 2760

acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc 2820acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc 2820

cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc 2880cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc 2880

caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg 2940caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg 2940

tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac 3000tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac 3000

cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg 3060cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg 3060

gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac 3120gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac 3120

ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc 3180ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc 3180

ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg 3240ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg 3240

cggctgtgcg aacgcattct ggcgatgcat gcgatcacca ctcttggttc gcaagtgtct 3300cggctgtgcg aacgcattct ggcgatgcat gcgatcacca ctcttggttc gcaagtgtct 3300

acacagcgct ccggttctta cgaaaactca aactcagcca ctgagggttc taccataaac 3360acacagcgct ccggttctta cgaaaactca aactcagcca ctgagggttc taccataaac 3360

tacaccacca ttaattacta caaagactcc tatgctgcca cagcaggcaa acagagtctc 3420tacaccacca ttaattacta caaagactcc tatgctgcca cagcaggcaa acagagtctc 3420

aagcaggatc cagacaagtt tgcaaatcct gttaaagaca tattcaccga aatggcagcg 3480aagcaggatc cagacaagtt tgcaaatcct gttaaagaca tattcaccga aatggcagcg 3480

ccactgaagt ccccatccgc tgaggcatgt ggatacagtg atcgagtggc gcaattaact 3540ccactgaagt ccccatccgc tgaggcatgt ggatacagtg atcgagtggc gcaattaact 3540

attggcaact ccaccatcac gacgcaagaa gcggctaaca tcatagtcgg ctatggtgag 3600attggcaact ccaccatcac gacgcaagaa gcggctaaca tcatagtcgg ctatggtgag 3600

tggccttcct actgctcaga ttctgacgct acagcagtgg ataaaccaac gcgcccggat 3660tggccttcct actgctcaga ttctgacgct acagcagtgg ataaaccaac gcgcccggat 3660

gtttcagtga acaggtttta cacattggac actaaattgt gggagaaatc gtccaaggga 3720gtttcagtga acaggtttta cacattggac actaaattgt gggagaaatc gtccaaggga 3720

tggtactgga agttcccgga tgtgttaact gaaactgggg tttttgggca aaatgcacaa 3780tggtactgga agttcccgga tgtgttaact gaaactgggg tttttgggca aaatgcacaa 3780

ttccactacc tctaccgatc agggttctgc atccacgtgc agtgcaatgc cagtaaattc 3840ttccactacc tctaccgatc agggttctgc atccacgtgc agtgcaatgc cagtaaattc 3840

caccaaggag cactcctagt cgctgtccta ccagagtatg tcattgggac agtggcaggc 3900caccaaggag cactcctagt cgctgtccta ccagagtatg tcattgggac agtggcaggc 3900

ggtacaggga cggaagacac ccaccccccc tacaagcaga cccaacccgg cgccgatggt 3960ggtacaggga cggaagacac ccacccccccc tacaagcaga cccaacccgg cgccgatggt 3960

ttcgagttgc aacacccgta cgtgcttgat gctggcatcc caatatcaca gttaacagtg 4020ttcgagttgc aacacccgta cgtgcttgat gctggcatcc caatatcaca gttaacagtg 4020

tgcccacacc agtggattaa tttgaggacc aacaattgtg ctacaataat agtgccatac 4080tgcccacacc agtggattaa tttgaggacc aacaattgtg ctacaataat agtgccatac 4080

attaacgcac tgccttttga ttctgccttg aaccattgca actttggcct gttagttgtg 4140attaacgcac tgccttttga ttctgccttg aaccattgca actttggcct gttagttgtg 4140

cctattagcc cactagacta cgaccaagga gcaacgccag taatccctat aactatcaca 4200cctattagcc cactagacta cgaccaagga gcaacgccag taatccctat aactatcaca 4200

ttggccccaa tgtgctctga attcgcaggt cttaggcagg cagtcacgca agggttcccc 4260ttggccccaa tgtgctctga attcgcaggt cttaggcagg cagtcacgca agggttcccc 4260

accgagctaa aacctggcac aaatcaattt ttaaccaccg atgatggcgt ctcagcacct 4320accgagctaa aacctggcac aaatcaattt ttaaccaccg atgatggcgt ctcagcacct 4320

attctaccaa acttccaccc caccccgtgt atccacatac ctggtgaagt taggaacttg 4380attctaccaa acttccaccc caccccgtgt atccacatac ctggtgaagt taggaacttg 4380

ctagagttat gccaggtgga gaccattctg gaggttaaca atgtgcccac gaatgccact 4440ctagagttat gccaggtgga gaccattctg gaggttaaca atgtgcccac gaatgccact 4440

agcttaatgg agagactgcg cttcccggtc tcagcacaag cagggaaagg tgaactgtgt 4500agcttaatgg agagactgcg cttcccggtc tcagcacaag cagggaaagg tgaactgtgt 4500

gcggtgttta gagccgatcc tgggcgaaat ggaccatggc aatccacctt actgggccag 4560gcggtgttta gagccgatcc tgggcgaaat ggaccatggc aatccacctt actgggccag 4560

ttgtgcgggt actacaccca atggtcaggg tcattggaag tcaccttcat gtttactgga 4620ttgtgcgggt actacaccca atggtcaggg tcattggaag tcaccttcat gtttactgga 4620

tccttcatgg ctaccggcaa gatgctcata gcctatacac cgccaggggg tcctctgccc 4680tccttcatgg ctaccggcaa gatgctcata gcctatacac cgccaggggg tcctctgccc 4680

aaggaccggg cgaccgccat gttgggcacg cacgtcatct gggattttgg gctgcaatcg 4740aaggaccggg cgaccgccat gttgggcacg cacgtcatct gggattttgg gctgcaatcg 4740

tctgttaccc ttgtaatacc atggatcagt aacactcatt atagagcaca tgcccgagat 4800tctgttaccc ttgtaatacc atggatcagt aacactcatt atagagcaca tgcccgagat 4800

ggagtgtttg actattacac tacagggtta gtcagtatat ggtaccagac aaattacgtg 4860ggagtgtttg actattacac tacagggtta gtcagtatat ggtaccagac aaattacgtg 4860

gttccaatcg gtgcgcccaa cacagcctat ataatagcac tagcggcagc ccaaaagaac 4920gttccaatcg gtgcgcccaa cacagcctat ataatagcac tagcggcagc ccaaaagaac 4920

ttcactatga aattgtgcaa ggatgctagt gatatcctgc agacgggcac catccaggga 4980ttcactatga aattgtgcaa ggatgctagt gatatcctgc agacgggcac catccaggga 4980

gatagggtgg cagatgtaat tgaaagttcc ataggagata gcgtgagcag agccctcact 5040gatagggtgg cagatgtaat tgaaagttcc ataggagata gcgtgagcag agccctcact 5040

cacgctctac cagcacccac aggccaaaac acacaggtga gcagtcatcg actggataca 5100cacgctctac cagcacccac aggccaaaac acacaggtga gcagtcatcg actggataca 5100

ggcaaggttc cagcactcca agctgctgaa attggggcat catcaaatgc tagtgacgag 5160ggcaaggttc cagcactcca agctgctgaa attggggcat catcaaatgc tagtgacgag 5160

agcatgattg aaacacgttg tgttcttaac tcgcatagta cagctgagac cactcttgat 5220agcatgattg aaacacgttg tgttcttaac tcgcatagta cagctgagac cactcttgat 5220

agtttcttca gtagggcagg attagttgga gagatagatc tccctcttga gggcacaact 5280agtttcttca gtagggcagg attagttgga gagatagatc tccctcttga gggcacaact 5280

aacccaaatg gttatgccaa ctgggacata gatataacag gttacgcgca aatgcgtaga 5340aacccaaatg gttatgccaa ctgggacata gatataacag gttacgcgca aatgcgtaga 5340

aaggtagagc tattcaccta catgcgtttt gatgcagagt tcacttttgt tgcgtgcaca 5400aaggtagagc tattcaccta catgcgtttt gatgcagagt tcacttttgt tgcgtgcaca 5400

cccaccgggg aggttgtccc acaattgctc caatatatgt ttgtgccacc tggagcccct 5460cccaccgggg aggttgtccc acaattgctc caatatatgt ttgtgccacc tggagcccct 5460

aagccagatt ctagggaatc ccttgcatgg caaaccgcca ccaacccctc agtttttgtc 5520aagccagatt ctagggaatc ccttgcatgg caaaccgcca ccaacccctc agtttttgtc 5520

aagctgtcag accctccggc gcaggtttca gtgccattca tgtcacctgc gagtgcttat 5580aagctgtcag accctccggc gcaggtttca gtgccattca tgtcacctgc gagtgcttat 5580

caatggtttt atgacggata tcccacattc ggagaacaca aacaggagaa agaccttgaa 5640caatggtttt atgacggata tcccacattc ggagaacaca aacaggagaa agaccttgaa 5640

tacggggcat gtcctaataa catgatgggt acattctcag tgcggactgt ggggacctcc 5700tacggggcat gtcctaataa catgatgggt acattctcag tgcggactgt ggggacctcc 5700

aagtccaagt accctttagt ggttaggatt tacatgagaa tgaagcacgt cagggcgtgg 5760aagtccaagt accctttagt ggttaggatt tacatgagaa tgaagcacgt cagggcgtgg 5760

atacctcgcc cgatgcgcaa ccagaactac ctgttcaaag ccaacccaaa ttatgctggc 5820atacctcgcc cgatgcgcaa ccagaactac ctgttcaaag ccaacccaaa ttatgctggc 5820

aactctatta agccaactgg tgccagtcgc acagcgatca ccactcttgg gaaatttgga 5880aactctatta agccaactgg tgccagtcgc acagcgatca ccactcttgg gaaatttgga 5880

caacagtctg gggctattta tgtgggcaac tttagagtgg tcaaccgaca tcttgccacc 5940caacagtctg gggctattta tgtgggcaac tttagagtgg tcaaccgaca tcttgccacc 5940

cataatgatt gggcaaatct tgtttgggaa gacagctctc gcgacttgct cgtgtcatcc 6000cataatgatt gggcaaatct tgtttgggaa gacagctctc gcgacttgct cgtgtcatcc 6000

accactgccc aaggttgtga cacgattgcc cgttgcgatt gccagacagg ggtgtactac 6060accactgccc aaggttgtga cacgattgcc cgttgcgatt gccagacagg ggtgtactac 6060

tgtaactcga tgagaaaaca ctacccagtc agtttttcaa aacccagcct gatctatgta 6120tgtaactcga tgagaaaaca ctacccagtc agtttttcaa aacccagcct gatctatgta 6120

gaggctagcg agtattaccc agccaggtac caatcacatc tcatgctcgc acagggtcac 6180gaggctagcg agtattaccc agccaggtac caatcacatc tcatgctcgc acagggtcac 6180

tcggaacctg gtgattgcgg tggtatcctt aggtgccaac atggcgtcat cggcatagtg 6240tcggaacctg gtgattgcgg tggtatcctt aggtgccaac atggcgtcat cggcatagtg 6240

tctactggtg gcaatgggct cgttggcttt gcagacgtca gagacctctt gtggttagat 6300tctactggtg gcaatgggct cgttggcttt gcagacgtca gagacctctt gtggttagat 6300

gaagaagcta tggaacaggg cgtgtccgac tacattaagg gtctcggaga tgcttttgga 6360gaagaagcta tggaacaggg cgtgtccgac tacattaagg gtctcggaga tgcttttgga 6360

acaggcttca ctgacgcagt ctcaagggag gttgaagctc tcaagaacta tcttataggg 6420acaggcttca ctgacgcagt ctcaagggag gttgaagctc tcaagaacta tcttataggg 6420

tctgaaggag cagttgagaa aattttgaaa aatcttatta aactaatctc tgcactggtg 6480tctgaaggag cagttgagaa aattttgaaa aatcttatta aactaatctc tgcactggtg 6480

attgtgatca gaagtgatta cgacatggtt accctcactg caaccttagc gctgataggt 6540attgtgatca gaagtgatta cgacatggtt accctcactg caaccttagc gctgataggt 6540

tgtcatggca gtccttgggc ttggattaaa gccaaaacag cctccatctt aggtatccct 6600tgtcatggca gtccttgggc ttggattaaa gccaaaacag cctccatctt aggtatccct 6600

atcgcccaaa agcagagcgc ttcctggctc aagaagttca atgacatggc caacgccgct 6660atcgcccaaa agcagagcgc ttcctggctc aagaagttca atgacatggc caacgccgct 6660

aaggggttag agtgggtttc caacaagatc agcaaattta ttgattggct taaggagaaa 6720aaggggttag agtgggtttc caacaagatc agcaaattta ttgattggct taaggagaaa 6720

atagtaccag cagccaggga gaaggttgaa ttcctaaata acttgaaaca gctgccactg 6780atagtaccag cagccaggga gaaggttgaa ttcctaaata acttgaaaca gctgccactg 6780

ctagagaatc agatctcgaa cttggaacaa tctgctgctt cacaagagga ccttgaagtc 6840ctagagaatc agatctcgaa cttggaacaa tctgctgctt cacaagagga ccttgaagtc 6840

atgtttggga atgtgtcgta cctagctcac ttctgtcgca agtttcaacc gctatacgcc 6900atgtttggga atgtgtcgta cctagctcac ttctgtcgca agtttcaacc gctatacgcc 6900

acggaagcta aaagagtcta tgccctggag aagagaatga ataactatat gcagttcaag 6960acggaagcta aaagagtcta tgccctggag aagagaatga ataactatat gcagttcaag 6960

agcaaacacc gaattgaacc tgtatgtctc attattaggg gctcaccagg caccgggaag 7020agcaaacacc gaattgaacc tgtatgtctc attattaggg gctcaccagg caccgggaag 7020

tctctagcca ctggtattat tgctcgagca atcgctgata agtaccactc cagcgtgtac 7080tctctagcca ctggtattat tgctcgagca atcgctgata agtaccactc cagcgtgtac 7080

tcgctcccac cagacccgga tcattttgac ggttacaagc aacaggtggt tacagtgatg 7140tcgctcccac cagacccgga tcattttgac ggttacaagc aacaggtggt tacagtgatg 7140

gatgatttgt gtcaaaaccc cgatggtaag gatatgtcct tattctgtca aatggtatcc 7200gatgatttgt gtcaaaaccc cgatggtaag gatatgtcct tattctgtca aatggtatcc 7200

accgtagatt tcattccacc aatggcttct ctcgaggaga agggagtttc cttcacctct 7260accgtagatt tcattccacc aatggcttct ctcgaggaga agggagtttc cttcacctct 7260

aagtttgtca tcgcatccac taatgccagt aatatcatag taccaacagt gtctgattct 7320aagtttgtca tcgcatccac taatgccagt aatatcatag taccaacagt gtctgattct 7320

gacgctattc gccgcaggtt ctacatggac tgtgacattg aagtgacaga ctcgtacaaa 7380gacgctattc gccgcaggtt ctacatggac tgtgacattg aagtgacaga ctcgtacaaa 7380

acagatctag gtagactgga tgcagggcga gccgctaaac tgtgttctga aaataacact 7440acagatctag gtagactgga tgcagggcga gccgctaaac tgtgttctga aaataacact 7440

gcaaatttca aacgttgcag cccattagtg tgtgggaaag ccatccaact tagagataga 7500gcaaatttca aacgttgcag cccattagtg tgtgggaaag ccatccaact tagagataga 7500

aagtctaaag tcagatacag tgtggatacg gtggtttcag aacttattag ggaatacagc 7560aagtctaaag tcagatacag tgtggatacg gtggtttcag aacttattag ggaatacagc 7560

aataggtccg ccattggtaa cacaatcgag gctcttttcc aaggtccacc caagttcagg 7620aataggtccg ccattggtaa cacaatcgag gctcttttcc aaggtccacc caagttcagg 7620

ccaattagga ttagccttga agaaaaacca gccccagacg ctattagcga tctccttgct 7680ccaattagga ttagccttga agaaaaacca gccccagacg ctattagcga tctccttgct 7680

agtgtagata gtgaagaagt gcgccagtac tgcagggatc aaggctggat tattcctgaa 7740agtgtagata gtgaagaagt gcgccagtac tgcagggatc aaggctggat tattcctgaa 7740

gctcccacca atgtggagcg gcaccttaat agagcggtgc tcgtcatgca atccatcacc 7800gctcccacca atgtggagcg gcaccttaat agagcggtgc tcgtcatgca atccatcacc 7800

acagtagtgg cggttgtttc gttggtgtac gtcatctaca agctctttgc agggtttcag 7860acagtagtgg cggttgtttc gttggtgtac gtcatctaca agctctttgc agggtttcag 7860

ggtgcatatt ctggtgctcc taagcaagtg cttaagaaac ctgctcttcg cacagcaaca 7920ggtgcatatt ctggtgctcc taagcaagtg cttaagaaac ctgctcttcg cacagcaaca 7920

gtgcagggtc cgagccttga ctttgctctc tccctactga gaaggaacat caggcaggtc 7980gtgcagggtc cgagccttga ctttgctctc tccctactga gaaggaacat caggcaggtc 7980

caaacagacc aagggcattt caccatgttg ggtgttaggg atcgcttagc agtcctccca 8040caaacagacc aagggcattt caccatgttg ggtgttaggg atcgcttagc agtcctccca 8040

cgccactcac aacctggcaa aaccatttgg attgagcaca aactcgtgaa cgtccttgat 8100cgccactcac aacctggcaa aaccatttgg attgagcaca aactcgtgaa cgtccttgat 8100

gcagttgaac tggtggatga gcaaggagtc aacctggaat taaccctcat cactcttgac 8160gcagttgaac tggtggatga gcaaggagtc aacctggaat taaccctcat cactcttgac 8160

accaacgaga agtttaggga tatcaccaaa ttcatcccag aaaatatcag cactgctagc 8220accaacgaga agtttaggga tatcaccaaa ttcatcccag aaaatatcag cactgctagc 8220

gatgccaccc tagtgatcaa cacggagcac atgccgtcaa tgtttgtccc ggtgggtgac 8280gatgccaccc tagtgatcaa cacggagcac atgccgtcaa tgtttgtccc ggtgggtgac 8280

gttgtgcagt atggcttttt gaatctcagt ggcaagccta cccatcgcac catgatgtac 8340gttgtgcagt atggcttttt gaatctcagt ggcaagccta cccatcgcac catgatgtac 8340

aattttccta ctaaagcagg acagtgtgga ggagtggtga catctgttgg gaaggttgtc 8400aattttccta ctaaagcagg acagtgtgga ggagtggtga catctgttgg gaaggttgtc 8400

ggtattcaca ttggtggcaa tggcagacaa ggtttttgcg caggcctcaa aaggagttac 8460ggtattcaca ttggtggcaa tggcagacaa ggttttttgcg caggcctcaa aaggagttac 8460

tttgctagtg aacaaggaga gatccagtgg gttaagccca ataaagaaac tggaagactc 8520tttgctagtg aacaaggaga gatccagtgg gttaagccca ataaagaaac tggaagactc 8520

aacatcaatg gaccaacccg caccaagtta gaacctagtg tattccatga catcttcgag 8580aacatcaatg gaccaacccg caccaagtta gaacctagtg tattccatga catcttcgag 8580

ggaaataagg aaccagctgt cttgcacagt aaagaccccc gacttgaggt agattttgaa 8640ggaaataagg aaccagctgt cttgcacagt aaagaccccc gacttgaggt agattttgaa 8640

caggccctgt tctctaagta tgtgggaaac acactacatg agcctgacga gtacatcaaa 8700caggccctgt tctctaagta tgtgggaaac acactacatg agcctgacga gtacatcaaa 8700

gaggcagctc tacattatgc aaaccaatta aagcaactag aaatcaatac ctctcaaatg 8760gaggcagctc tacattatgc aaaccaatta aagcaactag aaatcaatac ctctcaaatg 8760

agcatggagg aggcctgcta tggtactgag aatcttgagg ctattgatct tcacactagt 8820agcatggagg aggcctgcta tggtactgag aatcttgagg ctattgatct tcacactagt 8820

gcaggttacc cctatagtgc cctagggata aagaaaagag acatcttaga ccctaccacc 8880gcaggttacc cctatagtgc cctagggata aagaaaagag acatcttaga ccctaccacc 8880

agggacgtga gtagaatgaa gttctacatg gacaagtatg gtcttgatct tccctactcc 8940agggacgtga gtagaatgaa gttctacatg gacaagtatg gtcttgatct tccctactcc 8940

acttatgtca aggacgagct acgctcgatt gataaaatca agaaagggaa gtcccgcctg 9000acttatgtca aggacgagct acgctcgatt gataaaatca agaaagggaa gtcccgcctg 9000

atcgaggcca gtagtctaaa tgattcagtg tacctcagaa tggctttcgg gcatttgtat 9060atcgaggcca gtagtctaaa tgattcagtg tacctcagaa tggctttcgg gcatttgtat 9060

gaggctttcc acgcaaatcc tgggacgata actggatcgg ccgtggggtg taaccctgac 9120gaggctttcc acgcaaatcc tgggacgata actggatcgg ccgtggggtg taaccctgac 9120

acattctgga gcaagctgcc aattttgctc cctggttcac tctttgcctt tgactactca 9180acattctgga gcaagctgcc aattttgctc cctggttcac tctttgcctt tgactactca 9180

ggctatgatg ccagccttag ccctgtctgg ttcagagcat tagaattggt tcttagggag 9240ggctatgatg ccagccttag ccctgtctgg ttcagagcat tagaattggt tcttagggag 9240

atagggtata gtgaagaggc aatctcactc attgagggaa tcaaccacac acatcatgtg 9300atagggtata gtgaagaggc aatctcactc attgagggaa tcaaccacac acatcatgtg 9300

tatcgtaata agacctattg cgtgcttggt gggatgccct caggctgttc aggaacatcc 9360tatcgtaata agacctattg cgtgcttggt gggatgccct caggctgttc aggaacatcc 9360

atcttcaact caatgatcaa caacattatt atcagagcac tgctcataaa aacatttaag 9420atcttcaact caatgatcaa caacattatt atcagagcac tgctcataaa aacatttaag 9420

ggcattgatt tggatgaact caacatggtc gcttatggag acgatgtgct cgctagctat 9480ggcattgatt tggatgaact caacatggtc gcttatggag acgatgtgct cgctagctat 9480

cccttcccaa ttgattgctt ggaactagca aagactggta aggagtatgg tctgaccatg 9540cccttcccaa ttgattgctt ggaactagca aagactggta aggagtatgg tctgaccatg 9540

acccctgctg ataaatctcc ttgctttaat gaggtcaatt ggggtaatgc gaccttcctc 9600acccctgctg ataaatctcc ttgctttaat gaggtcaatt ggggtaatgc gaccttcctc 9600

aaaaggggct ttttgcccga tgaacagttt ccatttttga ttcaccctac tatgccaatg 9660aaaaggggct ttttgcccga tgaacagttt ccatttttga ttcaccctac tatgccaatg 9660

agggagatcc atgagtccat tcgatggacc aaggacgcac ggaacactca agatcatgtg 9720agggagatcc atgagtccat tcgatggacc aaggacgcac ggaacactca agatcatgtg 9720

cggtccttgt gcctcctagc atggcataat ggtaagcaag aatacgagaa gtttgtgagc 9780cggtccttgt gcctcctagc atggcataat ggtaagcaag aatacgagaa gtttgtgagc 9780

acaattaggt ctgtcccagt agggagagcg ttggctattc caaattatga aaatcttaga 9840acaattaggt ctgtcccagt agggagagcg ttggctattc caaattatga aaatcttaga 9840

cgaaattggc tcgagttatt ttagaggtta tacacacctc aaccccacca gaaatctggt 9900cgaaattggc tcgagttatt ttagaggtta tacacacctc aaccccacca gaaatctggt 9900

cgtgaatgtg actggtgggg gtaaatttgt tataaccaga atagcaaaaa aaaaaaaaaa 9960cgtgaatgtg actggtgggg gtaaatttgt tataaccaga atagcaaaaa aaaaaaaaaa 9960

aaaaaaaaaa aaaaaaagct ta 9982aaaaaaaaaa aaaaaaagct ta 9982

<210> 6<210> 6

<211> 10187<211> 10187

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 6<400> 6

gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 60gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 60

tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 120tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 120

tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 180tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 180

atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 240atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 240

gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 300gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 300

aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 360aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 360

ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 420ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 420

tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 480tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 480

ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 540ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 540

ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 600ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 600

ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 660ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 660

gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 720gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 720

gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 780gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 780

gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct 840gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct 840

cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 900cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 900

acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 960acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 960

acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1020acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 1020

tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1080tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 1080

ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1140ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 1140

ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 1200ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 1200

tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 1260tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 1260

aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 1320aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt 1320

ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 1380ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 1380

ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 1440ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 1440

gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 1500gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 1500

gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 1560gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 1560

cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 1620cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga 1620

actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 1680actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 1680

ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 1740ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 1740

tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 1800tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 1800

ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 1860ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 1860

agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 1920agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 1920

aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt gtcgacgcgg 1980aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt gtcgacgcgg 1980

ccgctaatac gactcactat aggttaaaac agcctgtggg ttgcacccac tcacagggcc 2040ccgctaatac gactcactat aggttaaaac agcctgtggg ttgcacccac tcacagggcc 2040

tactgggcgc aagcactctg gtacctcggt acctttgtgc gcctgtttta cacccccccc 2100tactgggcgc aagcactctg gtacctcggt acctttgtgc gcctgtttta cacccccccc 2100

ccaatgaaac ttagaagcaa taaaccacga tcaatagcag gcataacgct ccagttatgt 2160ccaatgaaac ttagaagcaa taaaccacga tcaatagcag gcataacgct ccagttatgt 2160

cttgatcaag cacttctgtt tccccggact gagtatcaat agactgctcg cgcggttgaa 2220cttgatcaag cacttctgtt tccccggact gagtatcaat agactgctcg cgcggttgaa 2220

ggagaaaacg ttcgttatcc ggctaactac ttcggaaaac ctagtaacac catgaaagtt 2280ggagaaaacg ttcgttatcc ggctaactac ttcggaaaac ctagtaacac catgaaagtt 2280

gcggagagct tcgttcagca ctcccccagt gtagatcagg tcgatgagtc accgcgttcc 2340gcggagagct tcgttcagca ctcccccagt gtagatcagg tcgatgagtc accgcgttcc 2340

ccacgggcga ccgtggcggt ggctgcgttg gcggcctgcc catggggtaa cccatggggc 2400ccacgggcga ccgtggcggt ggctgcgttg gcggcctgcc catggggtaa cccatggggc 2400

gctctaatac ggacatggtg tgaagagtct actgagctag ttggtagtcc tccggcccct 2460gctctaatac ggacatggtg tgaagagtct actgagctag ttggtagtcc tccggcccct 2460

gaatgcggct aatcccaact gcggagcaca cgcccacaag ccagcgggta gtgtgtcgta 2520gaatgcggct aatcccaact gcggagcaca cgcccacaag ccagcgggta gtgtgtcgta 2520

acgggtaact ctgcagcgga accgactact ttgggtgtcc gtgtttcctt ttatctttat 2580acgggtaact ctgcagcgga accgactact ttgggtgtcc gtgtttcctt ttatctttat 2580

attggctgct tatggtgaca attaaagaat tgttaccata tagctattgg attagccatc 2640attggctgct tatggtgaca attaaagaat tgttaccata tagctattgg attagccatc 2640

cggtgtgcaa cagagcaatt atttacctat ttattggttt tgtaccatta acctcgaatt 2700cggtgtgcaa cagagcaatt atttacctat ttattggttt tgtaccatta acctcgaatt 2700

ctgtgaccac ccttaattat atcttgaccc ttaacacagc taaaccatat gatggtgagc 2760ctgtgaccac ccttaattat atcttgaccc ttaacacagc taaaccatat gatggtgagc 2760

aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta 2820aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta 2820

aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg 2880aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg 2880

accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc 2940accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc 2940

accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac 3000accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac 3000

ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac 3060ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac 3060

gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc 3120gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc 3120

atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag 3180atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag 3180

tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag 3240tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag 3240

gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac 3300gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac 3300

cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc 3360cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc 3360

acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag 3420acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag 3420

ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagat gcatgcgatc 3480ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagat gcatgcgatc 3480

accactcttg gttcgcaagt gtctacacag cgctccggtt cttacgaaaa ctcaaactca 3540accactcttg gttcgcaagt gtctacacag cgctccggtt cttacgaaaa ctcaaactca 3540

gccactgagg gttctaccat aaactacacc accattaatt actacaaaga ctcctatgct 3600gccactgagg gttctaccat aaactacacc accattaatt actacaaaga ctcctatgct 3600

gccacagcag gcaaacagag tctcaagcag gatccagaca agtttgcaaa tcctgttaaa 3660gccacagcag gcaaacagag tctcaagcag gatccagaca agtttgcaaa tcctgttaaa 3660

gacatattca ccgaaatggc agcgccactg aagtccccat ccgctgaggc atgtggatac 3720gacatattca ccgaaatggc agcgccactg aagtccccat ccgctgaggc atgtggatac 3720

agtgatcgag tggcgcaatt aactattggc aactccacca tcacgacgca agaagcggct 3780agtgatcgag tggcgcaatt aactattggc aactccacca tcacgacgca agaagcggct 3780

aacatcatag tcggctatgg tgagtggcct tcctactgct cagattctga cgctacagca 3840aacatcatag tcggctatgg tgagtggcct tcctactgct cagattctga cgctacagca 3840

gtggataaac caacgcgccc ggatgtttca gtgaacaggt tttacacatt ggacactaaa 3900gtggataaac caacgcgccc ggatgtttca gtgaacaggt tttacacatt ggacactaaa 3900

ttgtgggaga aatcgtccaa gggatggtac tggaagttcc cggatgtgtt aactgaaact 3960ttgtgggaga aatcgtccaa gggatggtac tggaagttcc cggatgtgtt aactgaaact 3960

ggggtttttg ggcaaaatgc acaattccac tacctctacc gatcagggtt ctgcatccac 4020ggggtttttg ggcaaaatgc acaattccac tacctctacc gatcagggtt ctgcatccac 4020

gtgcagtgca atgccagtaa attccaccaa ggagcactcc tagtcgctgt cctaccagag 4080gtgcagtgca atgccagtaa attccaccaa ggagcactcc tagtcgctgt cctaccagag 4080

tatgtcattg ggacagtggc aggcggtaca gggacggaag acacccaccc cccctacaag 4140tatgtcattg ggacagtggc aggcggtaca gggacggaag acacccaccc cccctacaag 4140

cagacccaac ccggcgccga tggtttcgag ttgcaacacc cgtacgtgct tgatgctggc 4200cagacccaac ccggcgccga tggtttcgag ttgcaacacc cgtacgtgct tgatgctggc 4200

atcccaatat cacagttaac agtgtgccca caccagtgga ttaatttgag gaccaacaat 4260atcccaatat cacagttaac agtgtgccca caccagtgga ttaatttgag gaccaacaat 4260

tgtgctacaa taatagtgcc atacattaac gcactgcctt ttgattctgc cttgaaccat 4320tgtgctacaa taatagtgcc atacattaac gcactgcctt ttgattctgc cttgaaccat 4320

tgcaactttg gcctgttagt tgtgcctatt agcccactag actacgacca aggagcaacg 4380tgcaactttg gcctgttagt tgtgcctatt agcccactag actacgacca aggagcaacg 4380

ccagtaatcc ctataactat cacattggcc ccaatgtgct ctgaattcgc aggtcttagg 4440ccagtaatcc ctataactat cacattggcc ccaatgtgct ctgaattcgc aggtcttagg 4440

caggcagtca cgcaagggtt ccccaccgag ctaaaacctg gcacaaatca atttttaacc 4500caggcagtca cgcaagggtt ccccaccgag ctaaaacctg gcacaaatca atttttaacc 4500

accgatgatg gcgtctcagc acctattcta ccaaacttcc accccacccc gtgtatccac 4560accgatgatg gcgtctcagc acctattcta ccaaacttcc accccacccc gtgtatccac 4560

atacctggtg aagttaggaa cttgctagag ttatgccagg tggagaccat tctggaggtt 4620atacctggtg aagttaggaa cttgctagag ttatgccagg tggagaccat tctggaggtt 4620

aacaatgtgc ccacgaatgc cactagctta atggagagac tgcgcttccc ggtctcagca 4680aacaatgtgc ccacgaatgc cactagctta atggagagac tgcgcttccc ggtctcagca 4680

caagcaggga aaggtgaact gtgtgcggtg tttagagccg atcctgggcg aaatggacca 4740caagcaggga aaggtgaact gtgtgcggtg tttagagccg atcctgggcg aaatggacca 4740

tggcaatcca ccttactggg ccagttgtgc gggtactaca cccaatggtc agggtcattg 4800tggcaatcca ccttactggg ccagttgtgc gggtactaca cccaatggtc agggtcattg 4800

gaagtcacct tcatgtttac tggatccttc atggctaccg gcaagatgct catagcctat 4860gaagtcacct tcatgtttac tggatccttc atggctaccg gcaagatgct catagcctat 4860

acaccgccag ggggtcctct gcccaaggac cgggcgaccg ccatgttggg cacgcacgtc 4920acaccgccag ggggtcctct gcccaaggac cgggcgaccg ccatgttggg cacgcacgtc 4920

atctgggatt ttgggctgca atcgtctgtt acccttgtaa taccatggat cagtaacact 4980atctgggatt ttgggctgca atcgtctgtt acccttgtaa taccatggat cagtaacact 4980

cattatagag cacatgcccg agatggagtg tttgactatt acactacagg gttagtcagt 5040cattatagag cacatgcccg agatggagtg tttgactatt acactacagg gttagtcagt 5040

atatggtacc agacaaatta cgtggttcca atcggtgcgc ccaacacagc ctatataata 5100atatggtacc agacaaatta cgtggttcca atcggtgcgc ccaacacagc ctatataata 5100

gcactagcgg cagcccaaaa gaacttcact atgaaattgt gcaaggatgc tagtgatatc 5160gcactagcgg cagcccaaaa gaacttcact atgaaattgt gcaaggatgc tagtgatatc 5160

ctgcagacgg gcaccatcca gggagatagg gtggcagatg taattgaaag ttccatagga 5220ctgcagacgg gcaccatcca gggagatagg gtggcagatg taattgaaag ttccatagga 5220

gatagcgtga gcagagccct cactcacgct ctaccagcac ccacaggcca aaacacacag 5280gatagcgtga gcagagccct cactcacgct ctaccagcac ccacaggcca aaacacacag 5280

gtgagcagtc atcgactgga tacaggcaag gttccagcac tccaagctgc tgaaattggg 5340gtgagcagtc atcgactgga tacaggcaag gttccagcac tccaagctgc tgaaattggg 5340

gcatcatcaa atgctagtga cgagagcatg attgaaacac gttgtgttct taactcgcat 5400gcatcatcaa atgctagtga cgagagcatg attgaaacac gttgtgttct taactcgcat 5400

agtacagctg agaccactct tgatagtttc ttcagtaggg caggattagt tggagagata 5460agtacagctg agaccactct tgatagtttc ttcagtaggg caggattagt tggagagata 5460

gatctccctc ttgagggcac aactaaccca aatggttatg ccaactggga catagatata 5520gatctccctc ttgagggcac aactaaccca aatggttatg ccaactggga catagatata 5520

acaggttacg cgcaaatgcg tagaaaggta gagctattca cctacatgcg ttttgatgca 5580acaggttacg cgcaaatgcg tagaaaggta gagctattca cctacatgcg ttttgatgca 5580

gagttcactt ttgttgcgtg cacacccacc ggggaggttg tcccacaatt gctccaatat 5640gagttcactt ttgttgcgtg cacacccacc ggggaggttg tcccacaatt gctccaatat 5640

atgtttgtgc cacctggagc ccctaagcca gattctaggg aatcccttgc atggcaaacc 5700atgtttgtgc cacctggagc ccctaagcca gattctaggg aatcccttgc atggcaaacc 5700

gccaccaacc cctcagtttt tgtcaagctg tcagaccctc cggcgcaggt ttcagtgcca 5760gccaccaacc cctcagtttt tgtcaagctg tcagaccctc cggcgcaggt ttcagtgcca 5760

ttcatgtcac ctgcgagtgc ttatcaatgg ttttatgacg gatatcccac attcggagaa 5820ttcatgtcac ctgcgagtgc ttatcaatgg ttttatgacg gatatcccac attcggagaa 5820

cacaaacagg agaaagacct tgaatacggg gcatgtccta ataacatgat gggtacattc 5880cacaaacagg agaaagacct tgaatacggg gcatgtccta ataacatgat gggtacattc 5880

tcagtgcgga ctgtggggac ctccaagtcc aagtaccctt tagtggttag gatttacatg 5940tcagtgcgga ctgtggggac ctccaagtcc aagtaccctt tagtggttag gatttacatg 5940

agaatgaagc acgtcagggc gtggatacct cgcccgatgc gcaaccagaa ctacctgttc 6000agaatgaagc acgtcagggc gtggatacct cgcccgatgc gcaaccagaa ctacctgttc 6000

aaagccaacc caaattatgc tggcaactct attaagccaa ctggtgccag tcgcacagcg 6060aaagccaacc caaattatgc tggcaactct attaagccaa ctggtgccag tcgcacagcg 6060

atcaccactc ttgggaaatt tggacaacag tctggggcta tttatgtggg caactttaga 6120atcaccactc ttgggaaatt tggacaacag tctggggcta tttatgtggg caactttaga 6120

gtggtcaacc gacatcttgc cacccataat gattgggcaa atcttgtttg ggaagacagc 6180gtggtcaacc gacatcttgc cacccataat gattgggcaa atcttgtttg ggaagacagc 6180

tctcgcgact tgctcgtgtc atccaccact gcccaaggtt gtgacacgat tgcccgttgc 6240tctcgcgact tgctcgtgtc atccaccact gcccaaggtt gtgacacgat tgcccgttgc 6240

gattgccaga caggggtgta ctactgtaac tcgatgagaa aacactaccc agtcagtttt 6300gattgccaga caggggtgta ctactgtaac tcgatgagaa aacactaccc agtcagtttt 6300

tcaaaaccca gcctgatcta tgtagaggct agcgagtatt acccagccag gtaccaatca 6360tcaaaaccca gcctgatcta tgtagaggct agcgagtatt acccagccag gtaccaatca 6360

catctcatgc tcgcacaggg tcactcggaa cctggtgatt gcggtggtat ccttaggtgc 6420catctcatgc tcgcacaggg tcactcggaa cctggtgatt gcggtggtat ccttaggtgc 6420

caacatggcg tcatcggcat agtgtctact ggtggcaatg ggctcgttgg ctttgcagac 6480caacatggcg tcatcggcat agtgtctact ggtggcaatg ggctcgttgg ctttgcagac 6480

gtcagagacc tcttgtggtt agatgaagaa gctatggaac agggcgtgtc cgactacatt 6540gtcagagacc tcttgtggtt agatgaagaa gctatggaac agggcgtgtc cgactacatt 6540

aagggtctcg gagatgcttt tggaacaggc ttcactgacg cagtctcaag ggaggttgaa 6600aagggtctcg gagatgcttt tggaacaggc ttcactgacg cagtctcaag ggaggttgaa 6600

gctctcaaga actatcttat agggtctgaa ggagcagttg agaaaatttt gaaaaatctt 6660gctctcaaga actatcttat agggtctgaa ggagcagttg agaaaatttt gaaaaatctt 6660

attaaactaa tctctgcact ggtgattgtg atcagaagtg attacgacat ggttaccctc 6720attaaactaa tctctgcact ggtgattgtg atcagaagtg attacgacat ggttaccctc 6720

actgcaacct tagcgctgat aggttgtcat ggcagtcctt gggcttggat taaagccaaa 6780actgcaacct tagcgctgat aggttgtcat ggcagtcctt gggcttggat taaagccaaa 6780

acagcctcca tcttaggtat ccctatcgcc caaaagcaga gcgcttcctg gctcaagaag 6840acagcctcca tcttaggtat ccctatcgcc caaaagcaga gcgcttcctg gctcaagaag 6840

ttcaatgaca tggccaacgc cgctaagggg ttagagtggg tttccaacaa gatcagcaaa 6900ttcaatgaca tggccaacgc cgctaagggg ttagagtggg tttccaacaa gatcagcaaa 6900

tttattgatt ggcttaagga gaaaatagta ccagcagcca gggagaaggt tgaattccta 6960tttattgatt ggcttaagga gaaaatagta ccagcagcca gggagaaggt tgaattccta 6960

aataacttga aacagctgcc actgctagag aatcagatct cgaacttgga acaatctgct 7020aataacttga aacagctgcc actgctagg aatcagatct cgaacttgga acaatctgct 7020

gcttcacaag aggaccttga agtcatgttt gggaatgtgt cgtacctagc tcacttctgt 7080gcttcacaag aggaccttga agtcatgttt gggaatgtgt cgtacctagc tcacttctgt 7080

cgcaagtttc aaccgctata cgccacggaa gctaaaagag tctatgccct ggagaagaga 7140cgcaagtttc aaccgctata cgccacggaa gctaaaagag tctatgccct ggagaagaga 7140

atgaataact atatgcagtt caagagcaaa caccgaattg aacctgtatg tctcattatt 7200atgaataact atatgcagtt caagagcaaa caccgaattg aacctgtatg tctcattatt 7200

aggggctcac caggcaccgg gaagtctcta gccactggta ttattgctcg agcaatcgct 7260aggggctcac caggcaccgg gaagtctcta gccactggta ttattgctcg agcaatcgct 7260

gataagtacc actccagcgt gtactcgctc ccaccagacc cggatcattt tgacggttac 7320gataagtacc actccagcgt gtactcgctc ccaccagacc cggatcattt tgacggttac 7320

aagcaacagg tggttacagt gatggatgat ttgtgtcaaa accccgatgg taaggatatg 7380aagcaacagg tggttacagt gatggatgat ttgtgtcaaa accccgatgg taaggatg 7380

tccttattct gtcaaatggt atccaccgta gatttcattc caccaatggc ttctctcgag 7440tccttattct gtcaaatggt atccaccgta gatttcattc caccaatggc ttctctcgag 7440

gagaagggag tttccttcac ctctaagttt gtcatcgcat ccactaatgc cagtaatatc 7500gagaagggag tttccttcac ctctaagttt gtcatcgcat ccactaatgc cagtaatatc 7500

atagtaccaa cagtgtctga ttctgacgct attcgccgca ggttctacat ggactgtgac 7560atagtaccaa cagtgtctga ttctgacgct attcgccgca ggttctacat ggactgtgac 7560

attgaagtga cagactcgta caaaacagat ctaggtagac tggatgcagg gcgagccgct 7620attgaagtga cagactcgta caaaacagat ctaggtagac tggatgcagg gcgagccgct 7620

aaactgtgtt ctgaaaataa cactgcaaat ttcaaacgtt gcagcccatt agtgtgtggg 7680aaactgtgtt ctgaaaataa cactgcaaat ttcaaacgtt gcagcccatt agtgtgtgggg 7680

aaagccatcc aacttagaga tagaaagtct aaagtcagat acagtgtgga tacggtggtt 7740aaagccatcc aacttagaga tagaaagtct aaagtcagat acagtgtgga tacggtggtt 7740

tcagaactta ttagggaata cagcaatagg tccgccattg gtaacacaat cgaggctctt 7800tcagaactta ttagggaata cagcaatagg tccgccattg gtaacacaat cgaggctctt 7800

ttccaaggtc cacccaagtt caggccaatt aggattagcc ttgaagaaaa accagcccca 7860ttccaaggtc cacccaagtt caggccaatt aggattagcc ttgaagaaaa accagcccca 7860

gacgctatta gcgatctcct tgctagtgta gatagtgaag aagtgcgcca gtactgcagg 7920gacgctatta gcgatctcct tgctagtgta gatagtgaag aagtgcgcca gtactgcagg 7920

gatcaaggct ggattattcc tgaagctccc accaatgtgg agcggcacct taatagagcg 7980gatcaaggct ggattattcc tgaagctccc accaatgtgg agcggcacct taatagagcg 7980

gtgctcgtca tgcaatccat caccacagta gtggcggttg tttcgttggt gtacgtcatc 8040gtgctcgtca tgcaatccat caccacagta gtggcggttg tttcgttggt gtacgtcatc 8040

tacaagctct ttgcagggtt tcagggtgca tattctggtg ctcctaagca agtgcttaag 8100tacaagctct ttgcagggtt tcagggtgca tattctggtg ctcctaagca agtgcttaag 8100

aaacctgctc ttcgcacagc aacagtgcag ggtccgagcc ttgactttgc tctctcccta 8160aaacctgctc ttcgcacagc aacagtgcag ggtccgagcc ttgactttgc tctctcccta 8160

ctgagaagga acatcaggca ggtccaaaca gaccaagggc atttcaccat gttgggtgtt 8220ctgagaagga acatcaggca ggtccaaaca gaccaagggc atttcaccat gttgggtgtt 8220

agggatcgct tagcagtcct cccacgccac tcacaacctg gcaaaaccat ttggattgag 8280agggatcgct tagcagtcct cccacgccac tcacaacctg gcaaaaccat ttggattgag 8280

cacaaactcg tgaacgtcct tgatgcagtt gaactggtgg atgagcaagg agtcaacctg 8340cacaaactcg tgaacgtcct tgatgcagtt gaactggtgg atgagcaagg agtcaacctg 8340

gaattaaccc tcatcactct tgacaccaac gagaagttta gggatatcac caaattcatc 8400gaattaaccc tcatcactct tgacaccaac gagaagttta gggatatcac caaattcatc 8400

ccagaaaata tcagcactgc tagcgatgcc accctagtga tcaacacgga gcacatgccg 8460ccagaaaata tcagcactgc tagcgatgcc accctagtga tcaacacgga gcacatgccg 8460

tcaatgtttg tcccggtggg tgacgttgtg cagtatggct ttttgaatct cagtggcaag 8520tcaatgtttg tcccggtggg tgacgttgtg cagtatggct ttttgaatct cagtggcaag 8520

cctacccatc gcaccatgat gtacaatttt cctactaaag caggacagtg tggaggagtg 8580cctacccatc gcaccatgat gtacaatttt cctactaaag caggacagtg tggaggagtg 8580

gtgacatctg ttgggaaggt tgtcggtatt cacattggtg gcaatggcag acaaggtttt 8640gtgacatctg ttgggaaggt tgtcggtatt cacattggtg gcaatggcag acaaggtttt 8640

tgcgcaggcc tcaaaaggag ttactttgct agtgaacaag gagagatcca gtgggttaag 8700tgcgcaggcc tcaaaaggag ttactttgct agtgaacaag gagagatcca gtgggttaag 8700

cccaataaag aaactggaag actcaacatc aatggaccaa cccgcaccaa gttagaacct 8760cccaataaag aaactggaag actcaacatc aatggaccaa cccgcaccaa gttagaacct 8760

agtgtattcc atgacatctt cgagggaaat aaggaaccag ctgtcttgca cagtaaagac 8820agtgtattcc atgacatctt cgagggaaat aaggaaccag ctgtcttgca cagtaaagac 8820

ccccgacttg aggtagattt tgaacaggcc ctgttctcta agtatgtggg aaacacacta 8880ccccgacttg aggtagattt tgaacaggcc ctgttctcta agtatgtggg aaacacacta 8880

catgagcctg acgagtacat caaagaggca gctctacatt atgcaaacca attaaagcaa 8940catgagcctg acgagtacat caaagaggca gctctacatt atgcaaacca attaaagcaa 8940

ctagaaatca atacctctca aatgagcatg gaggaggcct gctatggtac tgagaatctt 9000ctagaaatca atacctctca aatgagcatg gaggaggcct gctatggtac tgagaatctt 9000

gaggctattg atcttcacac tagtgcaggt tacccctata gtgccctagg gataaagaaa 9060gaggctattg atcttcacac tagtgcaggt tacccctata gtgccctagg gataaagaaa 9060

agagacatct tagaccctac caccagggac gtgagtagaa tgaagttcta catggacaag 9120agagacatct tagaccctac caccagggac gtgagtagaa tgaagttcta catggacaag 9120

tatggtcttg atcttcccta ctccacttat gtcaaggacg agctacgctc gattgataaa 9180tatggtcttg atcttcccta ctccacttat gtcaaggacg agctacgctc gattgataaa 9180

atcaagaaag ggaagtcccg cctgatcgag gccagtagtc taaatgattc agtgtacctc 9240atcaagaaag ggaagtcccg cctgatcgag gccagtagtc taaatgattc agtgtacctc 9240

agaatggctt tcgggcattt gtatgaggct ttccacgcaa atcctgggac gataactgga 9300agaatggctt tcgggcattt gtatgaggct ttccacgcaa atcctgggac gataactgga 9300

tcggccgtgg ggtgtaaccc tgacacattc tggagcaagc tgccaatttt gctccctggt 9360tcggccgtgg ggtgtaaccc tgacacattc tggagcaagc tgccaatttt gctccctggt 9360

tcactctttg cctttgacta ctcaggctat gatgccagcc ttagccctgt ctggttcaga 9420tcactctttg cctttgacta ctcaggctat gatgccagcc ttagccctgt ctggttcaga 9420

gcattagaat tggttcttag ggagataggg tatagtgaag aggcaatctc actcattgag 9480gcattagaat tggttcttag ggagataggg tatagtgaag aggcaatctc actcattgag 9480

ggaatcaacc acacacatca tgtgtatcgt aataagacct attgcgtgct tggtgggatg 9540ggaatcaacc acacacatca tgtgtatcgt aataagacct attgcgtgct tggtgggatg 9540

ccctcaggct gttcaggaac atccatcttc aactcaatga tcaacaacat tattatcaga 9600ccctcaggct gttcaggaac atccatcttc aactcaatga tcaacaacat tattatcaga 9600

gcactgctca taaaaacatt taagggcatt gatttggatg aactcaacat ggtcgcttat 9660gcactgctca taaaaacatt taagggcatt gatttggatg aactcaacat ggtcgcttat 9660

ggagacgatg tgctcgctag ctatcccttc ccaattgatt gcttggaact agcaaagact 9720ggagacgatg tgctcgctag ctatcccttc ccaattgatt gcttggaact agcaaagact 9720

ggtaaggagt atggtctgac catgacccct gctgataaat ctccttgctt taatgaggtc 9780ggtaaggagt atggtctgac catgacccct gctgataaat ctccttgctt taatgaggtc 9780

aattggggta atgcgacctt cctcaaaagg ggctttttgc ccgatgaaca gtttccattt 9840aattggggta atgcgacctt cctcaaaagg ggctttttgc ccgatgaaca gtttccattt 9840

ttgattcacc ctactatgcc aatgagggag atccatgagt ccattcgatg gaccaaggac 9900ttgattcacc ctactatgcc aatgagggag atccatgagt ccattcgatg gaccaaggac 9900

gcacggaaca ctcaagatca tgtgcggtcc ttgtgcctcc tagcatggca taatggtaag 9960gcacggaaca ctcaagatca tgtgcggtcc ttgtgcctcc tagcatggca taatggtaag 9960

caagaatacg agaagtttgt gagcacaatt aggtctgtcc cagtagggag agcgttggct 10020caagaatacg agaagtttgt gagcacaatt aggtctgtcc cagtagggag agcgttggct 10020

attccaaatt atgaaaatct tagacgaaat tggctcgagt tattttagag gttatacaca 10080attccaaatt atgaaaatct tagacgaaat tggctcgagt tattttagag gttatacaca 10080

cctcaacccc accagaaatc tggtcgtgaa tgtgactggt gggggtaaat ttgttataac 10140cctcaacccc accagaaatc tggtcgtgaa tgtgactggt gggggtaaat ttgttataac 10140

cagaatagca aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agcttat 10187cagaatagca aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agcttat 10187

<210> 7<210> 7

<211> 19<211> 19

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 7<400> 7

taatacgact cactatagg 19taatacgact cactatagg 19

<210> 8<210> 8

<211> 30<211> 30

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 8<400> 8

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 30aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaa 30

<210> 9<210> 9

<211> 39<211> 39

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 9<400> 9

gctagcgctt tttttttttt tttttttttt ttttttttt 39gctagcgctt tttttttttt tttttttttt tttttttttt 39

<210> 10<210> 10

<211> 55<211> 55

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 10<400> 10

gacgcggccg ctaatacgac tcactatagg ttaaaacagc ctgtgggttg caccc 55gacgcggccg ctaatacgac tcactatagg ttaaaacagc ctgtgggttg caccc 55

<210> 11<210> 11

<211> 22<211> 22

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 11<400> 11

gcactgcacg tggatgcaga ac 22gcactgcacg tggatgcaga ac 22

<210> 12<210> 12

<211> 33<211> 33

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 12<400> 12

gacgcggccg cgttctgcat ccacgtgcag tgc 33gacgcggccg cgttctgcat ccacgtgcag tgc 33

<210> 13<210> 13

<211> 22<211> 22

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 13<400> 13

aagtcgcgag agctgtcttc cc 22aagtcgcgag agctgtcttc cc 22

<210> 14<210> 14

<211> 33<211> 33

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 14<400> 14

gacgcggccg cgggaagaca gctctcgcga ctt 33gacgcggccg cgggaagaca gctctcgcga ctt 33

<210> 15<210> 15

<211> 28<211> 28

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 15<400> 15

aattgtacat catggtgcga tgggtagg 28aattgtacat catggtgcga tgggtagg 28

<210> 16<210> 16

<211> 39<211> 39

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 16<400> 16

gacgcggccg ccctacccat cgcaccatga tgtacaatt 39gacgcggccg ccctacccat cgcaccatga tgtacaatt 39

<210> 17<210> 17

<211> 73<211> 73

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 17<400> 17

gctagcgctt tttttttttt tttttttttt tttttttttg ctattctggt tataacaaat 60gctagcgctt tttttttttt tttttttttt ttttttttttg ctattctggt tataacaaat 60

ttacccccac cag 73ttacccccac cag 73

<210> 18<210> 18

<211> 18<211> 18

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 18<400> 18

cctgacgtgt cgacgcgg 18cctgacgtgt cgacgcgg 18

<210> 19<210> 19

<211> 49<211> 49

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 19<400> 19

cctcgccctt gctcaccatc atatggttta gctgtgttaa gggtcaaga 49cctcgccctt gctcaccatc atatggttta gctgtgttaa gggtcaaga 49

<210> 20<210> 20

<211> 49<211> 49

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 20<400> 20

tcttgaccct taacacagct aaaccatatg atggtgagca agggcgagg 49tcttgaccct taacacagct aaaccatatg atggtgagca agggcgagg 49

<210> 21<210> 21

<211> 66<211> 66

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 21<400> 21

cgctgtgtag acacttgcga accaagagtg gtgatcgcat gcatcttgta cagctcgtcc 60cgctgtgtag acacttgcga accaagagtg gtgatcgcat gcatcttgta cagctcgtcc 60

atgccg 66atgccg 66

<210> 22<210> 22

<211> 66<211> 66

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 22<400> 22

cggcatggac gagctgtaca agatgcatgc gatcaccact cttggttcgc aagtgtctac 60cggcatggac gagctgtaca agatgcatgc gatcaccact cttggttcgc aagtgtctac 60

acagcg 66acagcg 66

<210> 23<210> 23

<211> 21<211> 21

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 23<400> 23

ctgcacgtgg atgcagaacc c 21ctgcacgtgg atgcagaacc c 21

<210> 24<210> 24

<211> 21<211> 21

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 24<400> 24

ctgcacgtgg atgcagaacc c 21ctgcacgtgg atgcagaacc c 21

<210> 25<210> 25

<211> 53<211> 53

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 25<400> 25

gaaatcttcg agtgtgaaga ccattctaga gtttagctgt gttaagggtc aag 53gaaatcttcg agtgtgaaga ccattctaga gtttagctgt gttaagggtc aag 53

<210> 26<210> 26

<211> 53<211> 53

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 26<400> 26

cttgaccctt aacacagcta aactctagaa tggtcttcac actcgaagat ttc 53cttgaccctt aacacagcta aactctagaa tggtcttcac actcgaagat ttc 53

<210> 27<210> 27

<211> 27<211> 27

<212> DNA<212> DNA

<213> Artificial<213> Artificial

<400> 27<400> 27

cgcatgcatc gccagaatgc gttcgca 27cgcatgcatc gccagaatgc gttcgca 27

Claims (20)

1. A cDNA, characterized in that it comprises the nucleic acid sequence of the EV71 strain and the nucleic acid sequence of a low copy plasmid backbone;
the nucleic acid sequence of the strain EV71 covers the 5 'to 3' forward polarity sequence of the EV71 virus, including the 5 'and 3' non-coding regions of the virus and one open reading frame encoding viral proteins.
2. The cDNA of claim 1, further comprising a sequence of a reporter gene, luciferase or a fluorescent protein, inserted in the nucleic acid sequence of the EV71 strain.
3. The cDNA according to claim 1, wherein the amino acid sequence of the open reading frame of the viral protein is as shown in SEQ ID NO 4.
4. The cDNA according to claim 1, wherein the coding sequence of the low copy plasmid backbone is as shown in SEQ ID NO 3.
5. The cDNA of claim 1, wherein the EV71 strain has a nucleic acid sequence shown in SEQ ID NO 2.
6. The cDNA according to claim 1, characterized in that its sequence is as shown in SEQ ID NO 1.
7. The expression product of the cDNA according to any one of claims 1 to 6.
8. A recombinant virus comprising a cDNA according to any one of claims 1 to 6.
9. A subgenomic replicon having a cDNA sequence according to any one of claims 1 to 6.
10. A double-stranded DNA capable of producing the cDNA according to any one of claims 1 to 6.
11. A plasmid containing the double-stranded DNA according to claim 10 or a derivative thereof.
12. The plasmid of claim 11, which is capable of transcribing to produce the full-length infectious RNA of the EV71 strain, or a mutant thereof.
13. A vaccine prepared from the plasmid of claim 11 or 12.
14. A viral vector, characterized in that it is prepared according to the plasmid of claim 11 or 12.
15. A viral particle produced from a cDNA clone according to any one of claims 1 to 6 or prepared from a plasmid according to claim 11 or 12.
16. A method for detecting EV71 virus, which comprises using the virus particle according to claim 15.
17. A method for producing an EV71 virus antibody, which comprises using the cDNA according to any one of claims 1 to 6 or the viral particle according to claim 15.
18. The method of claim 17, wherein the viral particle of claim 15 is used to immunize an animal and isolate an antibody, or to screen a human antibody library.
19. A kit for detecting EV71, comprising the cDNA of any one of claims 1 to 6 or the viral particle of claim 15.
20. Use of the cDNA of any one of claims 1 to 6 or the virion construct of claim 15 for the construction of a cell or animal model for further screening for a medicament against viral EV 71.
CN201910474088.3A 2019-06-02 2019-06-02 Infectious cDNA clone based on EV71 strain and application thereof Pending CN112094822A (en)

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