CN114085871A - 一种腺病毒质粒载体及腺病毒的制备方法 - Google Patents
一种腺病毒质粒载体及腺病毒的制备方法 Download PDFInfo
- Publication number
- CN114085871A CN114085871A CN202111471784.2A CN202111471784A CN114085871A CN 114085871 A CN114085871 A CN 114085871A CN 202111471784 A CN202111471784 A CN 202111471784A CN 114085871 A CN114085871 A CN 114085871A
- Authority
- CN
- China
- Prior art keywords
- adenovirus
- sequence
- plasmid vector
- seq
- site sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10111—Atadenovirus, e.g. ovine adenovirus D
- C12N2710/10121—Viruses as such, e.g. new isolates, mutants or their genomic sequences
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10111—Atadenovirus, e.g. ovine adenovirus D
- C12N2710/10141—Use of virus, viral particle or viral elements as a vector
- C12N2710/10143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10111—Atadenovirus, e.g. ovine adenovirus D
- C12N2710/10151—Methods of production or purification of viral material
- C12N2710/10152—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明提供了一种腺病毒质粒载体及腺病毒的制备方法,设计含有腺病毒质粒载体中20 bp的位点序列1和位点序列2的上下游引物,扩增获得目的基因片段,酶切获得末端具有同源序列的腺病毒质粒载体,通过Gibson DNA反应将目的基因组装入载体,载体中含有ccdB基因,极大地减少了重组子的筛选工作量,重组有效率95%以上,随后在包装细胞中制备腺病毒,减化了构建腺病毒的步骤,出毒成功率80%以上,病毒滴度大于1012 pfu/ml,并且保证了重组腺病毒基因组的完整性,腺病毒运载目的基因可在细胞内有效表达具有正常生物学活性的目的蛋白。本发明优化了腺病毒制备方法,在基于腺病毒的基因表达、疾病模型研究、基因治疗、疫苗研发等基础及临床转化研究领域具有广阔的应用前景。
Description
技术领域:
本发明属于分子生物学领域,具体涉及一种腺病毒质粒载体,及用腺病毒质粒载体制备腺病毒的方法。
技术背景:
腺病毒(Adenovirus, Ad)是一种非包膜线性双链DNA病毒,在人类有超过50种血清型腺病毒,腺病毒具有非整合特性、遗传稳定性、高基因转导效率以及易于大规模生产等优点,可作为基因传递载体,广泛用于表达各种外源基因、用于基因沉默的siRNAs或用于基因组编辑的CRISPR/Cas和设计核酸酶系统。其中人类腺病毒2型和5型是最常用的血清型,与病毒载体相比,腺病毒载体具有明显的优点。由于大多数哺乳动物细胞表达腺病毒一级受体和二级整合素受体,腺病毒是最有效和非整合的基因传递系统之一。腺病毒载体可通过修饰病毒衣壳,来优化腺病毒的治疗特征和靶向特异性。腺病毒载体已成为全球临床试验中最常用的病毒载体之一,在抗肿瘤、免疫治疗、疫苗接种和/或溶瘤治疗中发挥了重要作用。
尽管重组腺病毒载体的使用已经广泛,但腺病毒载体(尤其是第一代腺病毒载体)的初始构建和生产仍然是一个具有挑战性和耗时的过程。重组腺病毒载体通常通过四种方法产生。1)在表达腺病毒E1区的包装细胞(如HEK-293细胞)中将携带目的基因的腺病毒穿梭载体和腺病毒骨架基因组载体之间发生同源重组。2)使用独特的限制性内切酶将含有目的基因的DNA片段直接连接到线性化的E1/E3缺失的腺病毒基因组DNA片段。3)使用位点特异性重组酶和转座酶系统,如CRE/LOX和FLIP/FRT位点特异性整合和Gateway转座系统,将含有目的基因的DNA片段整合到腺病毒基因组DNA片段。4)利用微生物(如细菌和酵母)中更有效的同源重组反应来生成带有目的基因的腺病毒载体。
AdEasy腺病毒系统已经成为世界范围内生成腺病毒最常用的技术之一,但需要将目的基因先插入到穿梭质粒的多克隆区域,再与带有腺病毒骨架系统的AdEasy腺病毒骨架质粒在BJ5183大肠杆菌中进行同源重组获得腺病毒质粒。该过程需要两步法构建腺病毒质粒,且携带目的基因的阳性重组子的筛选过程需要挑取大量菌落进行鉴定,过程较复杂,整个制备时间约3-4周,出毒成功率较低,只有18-34%,且在BJ5183原核细胞中完成病毒基因组重组,病毒遗传背景及活性可能会受到影响。基于AdEasy腺病毒的制备方法在过去二十年中一直没有得到有效的改进。
发明内容:
为了解决检测现有技术中AdEasy腺病毒系统制备腺病毒过程复杂,时间长,效率低的问题,本发明提供了一种腺病毒质粒载体及腺病毒的优化制备方法,通过Gibson DNA组装(Gibson DNA Assembly, GDA)技术,一步法将目的基因导入腺病毒载体,操作简便,筛选效率高,腺病毒产率高。本发明的目的是通过以下措施实现的:
1.本发明提供了一种腺病毒质粒载体,包含5型腺病毒骨架基因序列,两个反向末端重复序列,启动子序列,多聚A尾序列,上述腺病毒质粒载体的启动子序列和多聚A尾序列之间有如SEQ NO.1所示的位点序列1及如SEQ NO.2所示的位点序列2,位点序列1靠近启动子序列,位点序列2靠近多聚A尾序列,在位点序列1及位点序列2之间由一个限制性核酸内切酶识别位点序列连接,该识别位点序列在腺病毒载体序列中是唯一的,经相应限制性内切酶酶切后产生平头末端,获得末端含有SEQ NO.1和SEQ NO.2的线性化腺病毒质粒载体片段;优选的,上述腺病毒质粒载体的位点序列1与位点序列2之间由限制性内切酶SwaІ的识别序列连接,连接后的序列如SEQ No3所示;
优选的,上述腺病毒质粒载体还含有绿色荧光蛋白或红色荧光蛋白的开放阅读框序列,所述开放阅读框序列位于另一启动子序列下游,用于编码绿色荧光蛋白或红色荧光蛋白;
优选的,上述腺病毒质粒载体的位点序列1与位点序列2之间连接有细菌自杀基因表达盒,
用于编码导致细菌死亡的毒素蛋白;
优选的,上述腺病毒质粒载体的位点序列1与位点序列2之间的细菌自杀基因序列为如S
EQ No.4所示的ccdB表达盒,表达盒两端分别有SwaI识别位点序列,载体序列如SEQ No.5所示;
优选的,上述SEQ No.6所示的腺病毒质粒载体中加入另一启动子序列及其下游的绿色荧光蛋白开放阅读框序列,载体序列如SEQ No.6所示;
优选的,上述SEQ No.6所示的腺病毒质粒载体中加入另一启动子序列及其下游的红色荧光蛋白开放阅读框序列,载体序列如SEQ No.7所示;
本发明还提供了一种一步法构建腺病毒的方法,包含以下步骤:
步骤1:设计5’端为SEQ NO.8的上游引物及5’端为SEQ NO.9的下游引物,使用高保真PCR试剂盒进行PCR扩增,获得目的基因片段;SEQ NO.8为SEQ NO.1及一半SwaI识别位点序列attt,SEQ NO.9为SEQ NO.2的反向互补序列及一半SwaI识别位点序列attt;
步骤2:用SwaI酶切腺病毒载体,获得末端与SEQ NO.8及SEQ NO.9同源的线性化载体片段;
步骤3:纯化后的目的基因片段与线性化载体片段混合,通过Gibson DNA组装,获得带有目的基因的重组腺病毒质粒;
步骤4:将带有目的基因的重组腺病毒质粒经PacI酶切线性化,转染到HEK-293或293A或293T腺病毒包装细胞中,5-7天收集细胞,裂解细胞获得腺病毒初提液;
步骤5:腺病毒初提液感染HEK-293或293A或293T细胞,进一步扩增,获得高滴度病毒液。
本发明还提供了一种腺病毒试剂盒,包含上述腺病毒质粒载体。
有益效果:
1. 本发明提供了一种腺病毒质粒载体,载体的启动子和多聚A尾之间包含细菌自杀基因ccdB,两侧的平头末端酶切识别位点序列及20 bp独特序列的位点序列1和位点序列2,它们是腺病毒基因组E1区GDA反应的通用位点,通过GDA反应,将末端具有相同位点序列1和位点序列2的DNA片段,组装入腺病毒质粒载体中,质粒载体带有的细菌自杀基因ccdB,可编码毒性蛋白,酶切不完全或自连环化的质粒均不能在LB平板上长出菌落,而重组后的腺病毒质粒没有细菌自杀基因ccdB,可在LB平板上长出菌落(图5),极大地减少了重组子的筛选工作量,阳性重组子有效率95%以上。
2. 这些本发明提供了一种腺病毒的优化制备方法,设计含有位点序列1和位点序列2位点的上下游引物,扩增获得目的基因片段,通过GDA组装将目的基因组装入腺病毒质粒载体中,一步法获得携带目的基因的腺病毒质粒中,随后在包装细胞中制备腺病毒。减化了构建腺病毒的步骤,简单方便,2-3周就能完成从质粒构建到重组出毒,出毒成功率80%以上,病毒滴度大于1012 pfu/ml,并且保证了重组腺病毒基因组的完整性,腺病毒运载目的基因可在细胞内有效表达具有正常生物学活性的目的蛋白。
3. 本发明还提供了一种制备腺病毒的试剂盒,大大简化了腺病毒制作过程,在基于腺病毒的基因表达、疾病模型研究、基因治疗、疫苗研发等基础及临床转化研究领域具有广阔的应用前景。
附图说明:
图1. 腺病毒质粒载体及腺病毒构建示意图(A. 腺病毒质粒载体;B. 腺病毒构建流程图)
图2. 腺病毒质粒载体pAdOSd图谱
图3. 腺病毒质粒载体pAdGOSd图谱
图4. 腺病毒质粒载体pAdROSd图谱
图5. 腺病毒质粒载体pAdGOSd和pAdGOS转化大肠杆菌DH10B后在Kan抗性平板上的菌落
图6. pAdOS-copGFP腺病毒质粒构建示意图
图7. pAdOS-copGFP腺病毒质粒构建过程(A. 菌落;B. 随机菌落PCR;C. 单酶切)
图8. AdOS-copGFP腺病毒构建过程(A. AdOS-copGFP腺病毒包装;B. AdOS-copGFP腺病毒滴度检测及扩增)
图9 . AdROS-mBMP9腺病毒构建过程(A. pAdROS-mBMP9腺病毒质粒构建AdROS-mBMP9;B. 菌落及随机菌落PCR;C. AdROS-mBMP9腺病毒包装;D. AdROS-mBMP9腺病毒感染小鼠骨髓间充质干细胞)
图10. AdROS-mBMP9对小鼠骨髓间充质干细胞的成骨作用(A-a. ALP染色;A-b.ALP活性;B. 茜素红染色;C. AdROS-mBMP9及AdRFP腺病毒感染小鼠骨髓间充质干细胞;D.皮下异位成骨组织的H.E及Masson染色)
具体实施方式:
下面通过实施例对本发明进行具体描述,有必要在此指出的是,以下实施例只用于对本发明进行进一步的说明,不能理解为对本发明保护范围的限制,该领域的技术熟练人员可以根据上述发明内容对本发明作出一些非本质的改进和调整。
实施例1. 一种腺病毒质粒载体系统
合成含有SEQ3的
5’-atccAATCGGAAAGCGGACGCGGAatttaaatCGAGTATCCCGTGAGCGCTTt-3’和含有SEQ3反向互补序列的
5’-ctagaAAGCGCTCACGGGATACTCGatttaaatTCCGCGTCCGCTTTCCGATTg-3’两条序列,体外退火后获得两端带有BamHI和XbaI 酶切末端的双链DNA,AdEasy系统的三个第一代腺病毒穿梭载体pShuttle CMV、pAdTrack CMV和pAdTrace CMV (Luo J, et al. A protocolfor rapid generation of recombinant adenoviruses using the AdEasy system. NatProtoc. 2007; 2(5):1236-47. doi: 10.1038/nprot.2007.135.)经BamHI和XbaI 酶切,双链DNA片段与酶切质粒载体经DNA连接酶连接,分别获得pShuttle MOS、pAdTrack MOS和pAdTrace MOS质粒载体。以上载体中的CMV-ployA表达盒中,被插入如SEQ No3所示DNA序列,由一个具有SwaI限制内切酶识别位点的序列atttaaat,位于该序列CMV启动子一侧的20bp位点序列1(modified one-step site 1, MOS1):AATCGGAAAGCGGACGCGGA(SEQ No.1),及位于该序列polyA一侧的位点序列2(modified one-step site 2, MOS2):GAGTATCCCGTGAGCGCTT(SEQ No.2)组成。
将pShuttle MOS、pAdTrack MOS和pAdTrace MOS穿梭质粒载体用PmeI限制性内切酶酶切线性化,并在BJ5183细菌中与pAdEasy1进行有效准确的同源重组反应。挑选卡那霉素抗性克隆,并通过PCR和限制性酶切和测序鉴定,获得腺病毒质粒载体,分别命名为pAdOS、pAdGOS和pAdROS。
用带有SwaI位点锚定的引物PCR扩增细菌自杀基因ccdB表达盒(SEQ No.4),连接到SwaI酶切消化的pAdOS载体中,并转化到有活性的DB3.1细菌中,通过PCR筛选菌落,提取阳性克隆菌落中的质粒,并通过PCR扩增、限制性酶切和DNA测序进一步验证。生成的GDA受体载体被命名为pAdOSd、pAdGOSd和pAdROSd。表S1中列出了所有寡核苷酸序列。pAdOSd质粒如图2所示,序列为SEQ No.5;pAdGOSd 质粒如图3所示,序列为SEQ No.6;pAdGOSd 质粒如图4所示,序列为SEQ No.7。
实施例2. 一种腺病毒构建方法
1)获得目的基因:设计扩增目的基因(gene of interest, GOI)的引物对,在上游引物的5’端加入如SEQ No.8所示序列AATCGGAAAGCGGACGCGGAattt,在下游引物的5’端加入如SEQ No.9所示序列AATCGGAAAGCGGACGCGGAattt,使用高保真PCR试剂盒进行PCR扩增,获得目的基因片段。以目的基因copGFP (copepoda中获得的绿色荧光蛋白GFP,亮度是EGFP的1.3倍,是GFP的7.8倍)为例,采用MOS1和kozak序列锚定的正向引物 SEQ No.10 和MOS2序列锚定的反向引物SEQ No.11,以pCDF1-MCS1-EF1-copGFP为模板进行扩增,获得含有copGFP编码基因,且末端分别为SEQ NO.8及SEQ NO.9的目的基因片段,经7.5M醋酸铵/乙醇沉淀,纯化后备用。
2)获得线性化载体片段:1-2μg腺病毒质粒载体pAdOSd或pAdGOSd或pAdROSd,2μlSwaI酶,5μl 10×酶切反应液,加ddH2O至50μl,25℃,酶切反应1-2小时,获得末端与SEQNO.8及SEQ NO.9同源的线性化载体片段;经7.5M醋酸铵/乙醇沉淀,纯化后备用。
3)Gibson DNA组装获得包含目的基因的腺病毒质粒:每个组装反应通常为10-15µl反应体积,包含上述目的基因片段100ng,线性化载体片段50ng,2×反应液5-7.5µl(NEB的Gibson Assembly Master Mix或NEBuilder HiFi DNA组装试剂盒),加ddH2O至10-15μl,50℃,GDA反应40~60分钟。通过7.5M醋酸铵/乙醇沉淀处理,纯化反应混合物。或者,将上述反应混合物加入至100µl的SwaI酶切反应体系,25℃,消化GDA反应混合物10分钟,然后用7.5M醋酸铵/乙醇沉淀处理。将纯化后的反应混合物重新悬浮在30µl ddH2O中,并使用15µl转化至感受态大肠杆菌克隆菌株,包括但不限于DH10B、Top10B、DH5α。向转化混合物中添加500µl LB,取100µl涂于LB/Kan平板上,37℃培养过夜。菌落PCR筛选阳性菌落,提取质粒,通过PCR、酶切及测序鉴定,获得携带目的基因的腺病毒质粒。另一条大片段
以目的基因copGFP 组装入腺病毒质粒载体pAdOSd为例(图6),约10%的GDA组装产物通过电转化DH10B,产生了上千个菌落(图7A)。随机挑选菌落进行菌落PCR筛选,16个克隆中有15个克隆含有copGFP的基因片段(图7B)。随机选取三个阳性克隆质粒,分别用四种限制性内切酶Hind III、Kpn I、Bam HI和Sph I酶切,并与腺病毒骨架载体pAdEasy1的酶切结果相比,电泳图表明三个阳性克隆的酶切片段大小均与预期一致(图7C),测序鉴定进一步证实三个阳性克隆中的copGFP 基因均正确组装至pAdOSd腺病毒质粒载体的位点序列1和位点序列2之间,获得pAdOS-copGFP腺病毒质粒。
该腺病毒质粒载体系统对于基于GDA的重组腺病毒质粒的快速构建是高效的,产生的pAdOS-copGFP质粒的腺病毒基因组结构稳定、完整性好。
4)包装并扩增腺病毒:将上述携带目的基因的重组腺病毒质粒经PacI酶切线性化,脂质体2000或3000按Invitrogen说明书转染,或聚乙烯亚胺(PEI)转染法,将上述线性化的重组腺病毒质粒转染到含有Ad5 E1区基因序列的腺病毒包装细胞中,腺病毒包装细胞包含但不限于HEK-293或293A或293T细胞。5-7天收集细胞,-80℃/37℃反复冻融四次,裂解细胞,获得腺病毒初提液,腺病毒初提液感染HEK-293或293A或293T细胞,进一步扩增,获得高滴度病毒液。
以copGFP腺病毒包装为例,pAdOS-copGFP腺病毒质粒经PacI酶切线性化,100μl酶切体系:3-5μg 腺病毒质粒DNA、3μl Pac I(NEB)、10×酶切反应液10μl,加ddH2O至100μl,37℃,酶切反应30分钟至1小时,凝胶电泳可见4.5kb的特异性小片段和另一条大片段,胶回收大片段于30μl ddH2O中,获得线性化的重组腺病毒质粒,线性DNA两端暴露出腺病毒反向末端重复序列(ITR)。转染前2~4小时,将在T-25培养瓶中培养HEK-293细胞,融合度30-40%,将30μl Pac I消化的DNA(含2-3μgDNA)和5~8μl PEI储备液(PEI浓度3μg/μl)(PEI:DNA=5-7μg :1μg)混合于200μl 无血清的DMEM培养基中,混合液室温放置10分钟。同时,从T-25培养瓶中小心吸出含10%FBS的完全培养基,(可选择清洗细胞一次:在培养瓶侧面加入5ml无血清DMEM,慢慢摇动使培养基覆盖细胞,然后吸出培养基),将上述DNA/PEI混合物加入到2.5ml无血清DMEM培养基中混合后,将加入烧瓶的一侧,轻轻摇动覆盖细胞,放置于37°C,5%CO2培养箱中。2-6小时后,去除培养基,换为8-10ml含10%FBS的DMEM完全培养基。转染后24小时观察GFP信号,评估转染效率,约10-15%细胞GFP阳性,GFP信号逐渐增强,第4天形成绿色彗星状荧光,表明腺病毒产生活跃并在局部扩增,荧光面积越来越大(图8A),第7天约有20-30%细胞变圆漂浮时,收集细胞沉淀,加1mlDMEM混悬细胞,-80℃及37℃反复冻融4次,离心取上清,获得病毒裂解液。用0.1%、0.5%、1%和5%四种不同滴度的病毒裂解液感染HEK-293细胞,荧光显微镜观察copGFP的表达水平,低至0.1%的病毒裂解物下可感染HEK-293细胞,且呈现剂量依赖性的表达模式(图8B)。10个pAdOS-copGFP中有8个能制备出表达copGPF的腺病毒,通过2~4轮HEK-293细胞反复感染获得高滴度AdOS-copGFP(>1012 pfu/ml)。以上结果表明该腺病毒质粒载体系统能高效构建重组腺病毒,出毒率为80%,并获得高滴度的腺病毒。
实施例3 腺病毒载体表达的小鼠BMP9的高成骨活性。
按照上述腺病毒制备方法,将小鼠骨形成发生蛋白9(mouse bone morphogeneticprotein, mBMP9)基因插入至AdROSd腺病毒质粒(图9A),GDA反应有效,并生成数百个阳性克隆,随机挑取16个菌落进行PCR检测,均携带mBMP9基因(图9B),随机挑取三个阳性克隆,经过酶切及测序鉴定证实三个阳性克隆中的mBMP9 基因均正确组装至pAdROSd腺病毒质粒载体的位点序列1和位点序列2之间,获得AdROS-mBMP9腺病毒质粒,经Pac I酶切线性化后,转染入HEK-293细胞中包装腺病毒(图9C),出毒率为90%,并经过2~4轮细胞反复感染扩增病毒,获得携带小鼠BMP9基因并表达红色荧光蛋白的高滴度(>1012 pfu/ml)腺病毒(AdROS-mBMP9),该病毒以滴度依赖的方式有效感染小鼠骨髓间充质干细胞(bone mesenchymalstem cells, BMSC)(图9D)。
进一步检测腺病毒运载的mBMP9的生物学功能,用AdROS-mBMP9和Ad-RFP对照病毒感染小鼠BMSC。与Ad-RFP对照组相比,AdROS-mBMP9组的碱性磷酸酶(ALP)染色阳性率(图10A-a)及ALP活性增高(图10A-b),且具有时间依赖性,茜素红染色阳性率在AdROS-mBMP9组也显著高于Ad-RFP对照组(图10B),表明腺病毒介导的mBMP9表达能够诱导小鼠骨髓间充质干细胞的早期成骨标志物ALP,促进骨基质矿化形成。
体内异位成骨实验:小鼠BMSC经AdROS-mBMP9或Ad-RFP感染16小时(图10C-a),收集细胞并重新悬浮在PBS中,进行裸鼠皮下注射(每注射点5×106细胞,每组5只,4-6周龄,雌性裸小鼠)。4周后通过H&E染色评估,AdROS-mBMP9组中检测到明显的骨形成,但RFP对照组中未检测到(图10C-b)。Masson三色染色进一步显示,腺病毒介导的mBMP9过表达能诱导小鼠BMSC在体内形成具有高度矿化骨基质的强健骨小梁(图10C-c)。以上结果表明,通过本发明的腺病毒制备方法,可很容易地将目的基因mBMP9导入腺病毒质粒载体中,由此产生的腺病毒可以有效地转导小鼠BMSC并诱导成骨分化。
序列表
<110> 重庆医科大学附属儿童医院
<120> 一种腺病毒质粒载体系统及一步法构建腺病毒的方法
<160> 11
<170> SIPOSequenceListing 1.0
<211> 20
<212> DNA
<213> Artificial(人工序列)
aatcggaaag cggacgcgga 20
<211> 20
<212> DNA
<213> Artificial(人工序列)
cgagtatccc gtgagcgctt 20
<211> 48
<212> DNA
<213> Artificial(人工序列)
aatcggaaag cggacgcgga atttaaatcg agtatcccgt gagcgctt 48
<211> 638
<212> DNA
<213> Artificial(人工序列 ddBd表达盒序列)
tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg 60
tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag 120
gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc 180
gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt 240
ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag 300
gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt 360
attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat 420
aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg 480
accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc 540
caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca 600
ggctccctta tacacagcca gtctgcagct cgctcttc 638
<211> 36344
<212> DNA
<213> Artificial(人工序列)
ctttaattaa ggatcccgac ctgtcctcga ccgatgccct tgagagcctt caacccagtc 60
agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt 120
atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc 180
tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc 240
ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt 300
atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc 360
tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg 420
caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc 480
gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc 540
gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc 600
tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc 660
ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga 720
gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca 780
gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg 840
tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg 900
aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag 960
caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag 1020
cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa 1080
cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc 1140
gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat 1200
catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga 1260
acagaaattc ccccttacac ggaggcatca agtgaccaaa caggaaaaaa ccgcccttaa 1320
catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga 1380
cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg 1440
cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga 1500
gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc 1560
agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt 1620
gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 1680
tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 1740
tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 1800
aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 1860
aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 1920
ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 1980
acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2040
ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2100
tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 2160
tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 2220
gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 2280
agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 2340
tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 2400
agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 2460
tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 2520
acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 2580
tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 2640
agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 2700
tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 2760
acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 2820
tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 2880
ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 2940
agtagttcgc cagttaatag tttgcgcaac gttgttgtca aaaaggatct tcacctagat 3000
ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 3060
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 3120
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 3180
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctc 3240
gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag gatgaggatc 3300
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 3360
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 3420
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 3480
tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 3540
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 3600
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 3660
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 3720
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 3780
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat 3840
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 3900
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 3960
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 4020
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 4080
ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc tcatttttta 4140
accaataggc cgaaatcggc aacatccctt ataaatcaaa agaatagacc gcgatagggt 4200
tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 4260
aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa 4320
gttttttgcg gtcgaggtgc cgtaaagctc taaatcggaa ccctaaaggg agcccccgat 4380
ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 4440
gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 4500
cgcgcttaat gcgccggatt aattaatcag tcccttccag ctctctgccc cttttggatt 4560
gaagccaata tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg 4620
gcgggtgacg tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa 4680
gcgacggatg tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa 4740
ttttcgcgcg gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg 4800
ccattttcgc gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca 4860
tagcgcgtaa tcgctagcag atcctggttc tttccgcctc agaagccata gagcccaccg 4920
catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc 4980
caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt 5040
aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa 5100
caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt tagcggccga 5160
tatcaagcgc tcacgggata ctcgatttaa attgattttt gcggtataag aatatatact 5220
gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga 5280
cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct 5340
ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga 5400
aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga 5460
cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt 5520
atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga 5580
tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg 5640
tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct 5700
ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca 5760
ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag 5820
ctcgctcttc atttaaattc cgcgtccgct ttccgattcc tgaattcggt gtcttctatg 5880
gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacagctcga 5940
tctctatcac tgatagggag atctctatca ctgataggga gatctctatc actgataggg 6000
agatctctat cactgatagg gagagctctg cttatataga cctcccaccg tacacgccta 6060
ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt 6120
tggttccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag 6180
tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg 6240
atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca 6300
taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga 6360
tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa 6420
tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg 6480
gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc 6540
atatatgggc tatgaactaa tgaccccgta attgattact attaataact agagatcgta 6600
agggtgggaa agaatatata aggtgggggt cttatgtagt tttgtatctg ttttgcagca 6660
gccgccgccg ccatgagcac caactcgttt gatggaagca ttgtgagctc atatttgaca 6720
acgcgcatgc ccccatgggc cggggtgcgt cagaatgtga tgggctccag cattgatggt 6780
cgccccgtcc tgcccgcaaa ctctactacc ttgacctacg agaccgtgtc tggaacgccg 6840
ttggagactg cagcctccgc cgccgcttca gccgctgcag ccaccgcccg cgggattgtg 6900
actgactttg ctttcctgag cccgcttgca agcagtgcag cttcccgttc atccgcccgc 6960
gatgacaagt tgacggctct tttggcacaa ttggattctt tgacccggga acttaatgtc 7020
gtttctcagc agctgttgga tctgcgccag caggtttctg ccctgaaggc ttcctcccct 7080
cccaatgcgg tttaaaacat aaataaaaaa ccagactctg tttggatttg gatcaagcaa 7140
gtgtcttgct gtctttattt aggggttttg cgcgcgcggt aggcccggga ccagcggtct 7200
cggtcgttga gggtcctgtg tattttttcc aggacgtggt aaaggtgact ctggatgttc 7260
agatacatgg gcataagccc gtctctgggg tggaggtagc accactgcag agcttcatgc 7320
tgcggggtgg tgttgtagat gatccagtcg tagcaggagc gctgggcgtg gtgcctaaaa 7380
atgtctttca gtagcaagct gattgccagg ggcaggccct tggtgtaagt gtttacaaag 7440
cggttaagct gggatgggtg catacgtggg gatatgagat gcatcttgga ctgtattttt 7500
aggttggcta tgttcccagc catatccctc cggggattca tgttgtgcag aaccaccagc 7560
acagtgtatc cggtgcactt gggaaatttg tcatgtagct tagaaggaaa tgcgtggaag 7620
aacttggaga cgcccttgtg acctccaaga ttttccatgc attcgtccat aatgatggca 7680
atgggcccac gggcggcggc ctgggcgaag atatttctgg gatcactaac gtcatagttg 7740
tgttccagga tgagatcgtc ataggccatt tttacaaagc gcgggcggag ggtgccagac 7800
tgcggtataa tggttccatc cggcccaggg gcgtagttac cctcacagat ttgcatttcc 7860
cacgctttga gttcagatgg ggggatcatg tctacctgcg gggcgatgaa gaaaacggtt 7920
tccggggtag gggagatcag ctgggaagaa agcaggttcc tgagcagctg cgacttaccg 7980
cagccggtgg gcccgtaaat cacacctatt accgggtgca actggtagtt aagagagctg 8040
cagctgccgt catccctgag caggggggcc acttcgttaa gcatgtccct gactcgcatg 8100
ttttccctga ccaaatccgc cagaaggcgc tcgccgccca gcgatagcag ttcttgcaag 8160
gaagcaaagt ttttcaacgg tttgagaccg tccgccgtag gcatgctttt gagcgtttga 8220
ccaagcagtt ccaggcggtc ccacagctcg gtcacctgct ctacggcatc tcgatccagc 8280
atatctcctc gtttcgcggg ttggggcggc tttcgctgta cggcagtagt cggtgctcgt 8340
ccagacgggc cagggtcatg tctttccacg ggcgcagggt cctcgtcagc gtagtctggg 8400
tcacggtgaa ggggtgcgct ccgggctgcg cgctggccag ggtgcgcttg aggctggtcc 8460
tgctggtgct gaagcgctgc cggtcttcgc cctgcgcgtc ggccaggtag catttgacca 8520
tggtgtcata gtccagcccc tccgcggcgt ggcccttggc gcgcagcttg cccttggagg 8580
aggcgccgca cgaggggcag tgcagacttt tgagggcgta gagcttgggc gcgagaaata 8640
ccgattccgg ggagtaggca tccgcgccgc aggccccgca gacggtctcg cattccacga 8700
gccaggtgag ctctggccgt tcggggtcaa aaaccaggtt tcccccatgc tttttgatgc 8760
gtttcttacc tctggtttcc atgagccggt gtccacgctc ggtgacgaaa aggctgtccg 8820
tgtccccgta tacagacttg agaggcctgt cctcgagcgg tgttccgcgg tcctcctcgt 8880
atagaaactc ggaccactct gagacaaagg ctcgcgtcca ggccagcacg aaggaggcta 8940
agtgggaggg gtagcggtcg ttgtccacta gggggtccac tcgctccagg gtgtgaagac 9000
acatgtcgcc ctcttcggca tcaaggaagg tgattggttt gtaggtgtag gccacgtgac 9060
cgggtgttcc tgaagggggg ctataaaagg gggtgggggc gcgttcgtcc tcactctctt 9120
ccgcatcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctga aaagcgggca 9180
tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata ttcacctggc 9240
ccgcggtgat gcctttgagg gtggccgcat ccatctggtc agaaaagaca atctttttgt 9300
tgtcaagctt ggtggcaaac gacccgtaga gggcgttgga cagcaacttg gcgatggagc 9360
gcagggtttg gtttttgtcg cgatcggcgc gctccttggc cgcgatgttt agctgcacgt 9420
attcgcgcgc aacgcaccgc cattcgggaa agacggtggt gcgctcgtcg ggcaccaggt 9480
gcacgcgcca accgcggttg tgcagggtga caaggtcaac gctggtggct acctctccgc 9540
gtaggcgctc gttggtccag cagaggcggc cgcccttgcg cgagcagaat ggcggtaggg 9600
ggtctagctg cgtctcgtcc ggggggtctg cgtccacggt aaagaccccg ggcagcaggc 9660
gcgcgtcgaa gtagtctatc ttgcatcctt gcaagtctag cgcctgctgc catgcgcggg 9720
cggcaagcgc gcgctcgtat gggttgagtg ggggacccca tggcatgggg tgggtgagcg 9780
cggaggcgta catgccgcaa atgtcgtaaa cgtagagggg ctctctgagt attccaagat 9840
atgtagggta gcatcttcca ccgcggatgc tggcgcgcac gtaatcgtat agttcgtgcg 9900
agggagcgag gaggtcggga ccgaggttgc tacgggcggg ctgctctgct cggaagacta 9960
tctgcctgaa gatggcatgt gagttggatg atatggttgg acgctggaag acgttgaagc 10020
tggcgtctgt gagacctacc gcgtcacgca cgaaggaggc gtaggagtcg cgcagcttgt 10080
tgaccagctc ggcggtgacc tgcacgtcta gggcgcagta gtccagggtt tccttgatga 10140
tgtcatactt atcctgtccc ttttttttcc acagctcgcg gttgaggaca aactcttcgc 10200
ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtaa gagcctagca 10260
tgtagaactg gttgacggcc tggtaggcgc agcatccctt ttctacgggt agcgcgtatg 10320
cctgcgcggc cttccggagc gaggtgtggg tgagcgcaaa ggtgtccctg accatgactt 10380
tgaggtactg gtatttgaag tcagtgtcgt cgcatccgcc ctgctcccag agcaaaaagt 10440
ccgtgcgctt tttggaacgc ggatttggca gggcgaaggt gacatcgttg aagagtatct 10500
ttcccgcgcg aggcataaag ttgcgtgtga tgcggaaggg tcccggcacc tcggaacggt 10560
tgttaattac ctgggcggcg agcacgatct cgtcaaagcc gttgatgttg tggcccacaa 10620
tgtaaagttc caagaagcgc gggatgccct tgatggaagg caatttttta agttcctcgt 10680
aggtgagctc ttcaggggag ctgagcccgt gctctgaaag ggcccagtct gcaagatgag 10740
ggttggaagc gacgaatgag ctccacaggt cacgggccat tagcatttgc aggtggtcgc 10800
gaaaggtcct aaactggcga cctatggcca ttttttctgg ggtgatgcag tagaaggtaa 10860
gcgggtcttg ttcccagcgg tcccatccaa ggttcgcggc taggtctcgc gcggcagtca 10920
ctagaggctc atctccgccg aacttcatga ccagcatgaa gggcacgagc tgcttcccaa 10980
aggcccccat ccaagtatag gtctctacat cgtaggtgac aaagagacgc tcggtgcgag 11040
gatgcgagcc gatcgggaag aactggatct cccgccacca attggaggag tggctattga 11100
tgtggtgaaa gtagaagtcc ctgcgacggg ccgaacactc gtgctggctt ttgtaaaaac 11160
gtgcgcagta ctggcagcgg tgcacgggct gtacatcctg cacgaggttg acctgacgac 11220
cgcgcacaag gaagcagagt gggaatttga gcccctcgcc tggcgggttt ggctggtggt 11280
cttctacttc ggctgcttgt ccttgaccgt ctggctgctc gaggggagtt acggtggatc 11340
ggaccaccac gccgcgcgag cccaaagtcc agatgtccgc gcgcggcggt cggagcttga 11400
tgacaacatc gcgcagatgg gagctgtcca tggtctggag ctcccgcggc gtcaggtcag 11460
gcgggagctc ctgcaggttt acctcgcata gacgggtcag ggcgcgggct agatccaggt 11520
gatacctaat ttccaggggc tggttggtgg cggcgtcgat ggcttgcaag aggccgcatc 11580
cccgcggcgc gactacggta ccgcgcggcg ggcggtgggc cgcgggggtg tccttggatg 11640
atgcatctaa aagcggtgac gcgggcgagc ccccggaggt agggggggct ccggacccgc 11700
cgggagaggg ggcaggggca cgtcggcgcc gcgcgcgggc aggagctggt gctgcgcgcg 11760
taggttgctg gcgaacgcga cgacgcggcg gttgatctcc tgaatctggc gcctctgcgt 11820
gaagacgacg ggcccggtga gcttgagcct gaaagagagt tcgacagaat caatttcggt 11880
gtcgttgacg gcggcctggc gcaaaatctc ctgcacgtct cctgagttgt cttgataggc 11940
gatctcggcc atgaactgct cgatctcttc ctcctggaga tctccgcgtc cggctcgctc 12000
cacggtggcg gcgaggtcgt tggaaatgcg ggccatgagc tgcgagaagg cgttgaggcc 12060
tccctcgttc cagacgcggc tgtagaccac gcccccttcg gcatcgcggg cgcgcatgac 12120
cacctgcgcg agattgagct ccacgtgccg ggcgaagacg gcgtagtttc gcaggcgctg 12180
aaagaggtag ttgagggtgg tggcggtgtg ttctgccacg aagaagtaca taacccagcg 12240
tcgcaacgtg gattcgttga tatcccccaa ggcctcaagg cgctccatgg cctcgtagaa 12300
gtccacggcg aagttgaaaa actgggagtt gcgcgccgac acggttaact cctcctccag 12360
aagacggatg agctcggcga cagtgtcgcg cacctcgcgc tcaaaggcta caggggcctc 12420
ttcttcttct tcaatctcct cttccataag ggcctcccct tcttcttctt ctggcggcgg 12480
tgggggaggg gggacacggc ggcgacgacg gcgcaccggg aggcggtcga caaagcgctc 12540
gatcatctcc ccgcggcgac ggcgcatggt ctcggtgacg gcgcggccgt tctcgcgggg 12600
gcgcagttgg aagacgccgc ccgtcatgtc ccggttatgg gttggcgggg ggctgccatg 12660
cggcagggat acggcgctaa cgatgcatct caacaattgt tgtgtaggta ctccgccgcc 12720
gagggacctg agcgagtccg catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa 12780
ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc ggcagcgggc ggcggtcggg 12840
gttgtttctg gcggaggtgc tgctgatgat gtaattaaag taggcggtct tgagacggcg 12900
gatggtcgac agaagcacca tgtccttggg tccggcctgc tgaatgcgca ggcggtcggc 12960
catgccccag gcttcgtttt gacatcggcg caggtctttg tagtagtctt gcatgagcct 13020
ttctaccggc acttcttctt ctccttcctc ttgtcctgca tctcttgcat ctatcgctgc 13080
ggcggcggcg gagtttggcc gtaggtggcg ccctcttcct cccatgcgtg tgaccccgaa 13140
gcccctcatc ggctgaagca gggctaggtc ggcgacaacg cgctcggcta atatggcctg 13200
ctgcacctgc gtgagggtag actggaagtc atccatgtcc acaaagcggt ggtatgcgcc 13260
cgtgttgatg gtgtaagtgc agttggccat aacggaccag ttaacggtct ggtgacccgg 13320
ctgcgagagc tcggtgtacc tgagacgcga gtaagccctc gagtcaaata cgtagtcgtt 13380
gcaagtccgc accaggtact ggtatcccac caaaaagtgc ggcggcggct ggcggtagag 13440
gggccagcgt agggtggccg gggctccggg ggcgagatct tccaacataa ggcgatgata 13500
tccgtagatg tacctggaca tccaggtgat gccggcggcg gtggtggagg cgcgcggaaa 13560
gtcgcggacg cggttccaga tgttgcgcag cggcaaaaag tgctccatgg tcgggacgct 13620
ctggccggtc aggcgcgcgc aatcgttgac gctctaccgt gcaaaaggag agcctgtaag 13680
cgggcactct tccgtggtct ggtggataaa ttcgcaaggg tatcatggcg gacgaccggg 13740
gttcgagccc cgtatccggc cgtccgccgt gatccatgcg gttaccgccc gcgtgtcgaa 13800
cccaggtgtg cgacgtcaga caacggggga gtgctccttt tggcttcctt ccaggcgcgg 13860
cggctgctgc gctagctttt ttggccactg gccgcgcgca gcgtaagcgg ttaggctgga 13920
aagcgaaagc attaagtggc tcgctccctg tagccggagg gttattttcc aagggttgag 13980
tcgcgggacc cccggttcga gtctcggacc ggccggactg cggcgaacgg gggtttgcct 14040
ccccgtcatg caagaccccg cttgcaaatt cctccggaaa cagggacgag cccctttttt 14100
gcttttccca gatgcatccg gtgctgcggc agatgcgccc ccctcctcag cagcggcaag 14160
agcaagagca gcggcagaca tgcagggcac cctcccctcc tcctaccgcg tcaggagggg 14220
cgacatccgc ggttgacgcg gcagcagatg gtgattacga acccccgcgg cgccgggccc 14280
ggcactacct ggacttggag gagggcgagg gcctggcgcg gctaggagcg ccctctcctg 14340
agcggtaccc aagggtgcag ctgaagcgtg atacgcgtga ggcgtacgtg ccgcggcaga 14400
acctgtttcg cgaccgcgag ggagaggagc ccgaggagat gcgggatcga aagttccacg 14460
cagggcgcga gctgcggcat ggcctgaatc gcgagcggtt gctgcgcgag gaggactttg 14520
agcccgacgc gcgaaccggg attagtcccg cgcgcgcaca cgtggcggcc gccgacctgg 14580
taaccgcata cgagcagacg gtgaaccagg agattaactt tcaaaaaagc tttaacaacc 14640
acgtgcgtac gcttgtggcg cgcgaggagg tggctatagg actgatgcat ctgtgggact 14700
ttgtaagcgc gctggagcaa aacccaaata gcaagccgct catggcgcag ctgttcctta 14760
tagtgcagca cagcagggac aacgaggcat tcagggatgc gctgctaaac atagtagagc 14820
ccgagggccg ctggctgctc gatttgataa acatcctgca gagcatagtg gtgcaggagc 14880
gcagcttgag cctggctgac aaggtggccg ccatcaacta ttccatgctt agcctgggca 14940
agttttacgc ccgcaagata taccataccc cttacgttcc catagacaag gaggtaaaga 15000
tcgaggggtt ctacatgcgc atggcgctga aggtgcttac cttgagcgac gacctgggcg 15060
tttatcgcaa cgagcgcatc cacaaggccg tgagcgtgag ccggcggcgc gagctcagcg 15120
accgcgagct gatgcacagc ctgcaaaggg ccctggctgg cacgggcagc ggcgatagag 15180
aggccgagtc ctactttgac gcgggcgctg acctgcgctg ggccccaagc cgacgcgccc 15240
tggaggcagc tggggccgga cctgggctgg cggtggcacc cgcgcgcgct ggcaacgtcg 15300
gcggcgtgga ggaatatgac gaggacgatg agtacgagcc agaggacggc gagtactaag 15360
cggtgatgtt tctgatcaga tgatgcaaga cgcaacggac ccggcggtgc gggcggcgct 15420
gcagagccag ccgtccggcc ttaactccac ggacgactgg cgccaggtca tggaccgcat 15480
catgtcgctg actgcgcgca atcctgacgc gttccggcag cagccgcagg ccaaccggct 15540
ctccgcaatt ctggaagcgg tggtcccggc gcgcgcaaac cccacgcacg agaaggtgct 15600
ggcgatcgta aacgcgctgg ccgaaaacag ggccatccgg cccgacgagg ccggcctggt 15660
ctacgacgcg ctgcttcagc gcgtggctcg ttacaacagc ggcaacgtgc agaccaacct 15720
ggaccggctg gtgggggatg tgcgcgaggc cgtggcgcag cgtgagcgcg cgcagcagca 15780
gggcaacctg ggctccatgg ttgcactaaa cgccttcctg agtacacagc ccgccaacgt 15840
gccgcgggga caggaggact acaccaactt tgtgagcgca ctgcggctaa tggtgactga 15900
gacaccgcaa agtgaggtgt accagtctgg gccagactat tttttccaga ccagtagaca 15960
aggcctgcag accgtaaacc tgagccaggc tttcaaaaac ttgcaggggc tgtggggggt 16020
gcgggctccc acaggcgacc gcgcgaccgt gtctagcttg ctgacgccca actcgcgcct 16080
gttgctgctg ctaatagcgc ccttcacgga cagtggcagc gtgtcccggg acacatacct 16140
aggtcacttg ctgacactgt accgcgaggc cataggtcag gcgcatgtgg acgagcatac 16200
tttccaggag attacaagtg tcagccgcgc gctggggcag gaggacacgg gcagcctgga 16260
ggcaacccta aactacctgc tgaccaaccg gcggcagaag atcccctcgt tgcacagttt 16320
aaacagcgag gaggagcgca ttttgcgcta cgtgcagcag agcgtgagcc ttaacctgat 16380
gcgcgacggg gtaacgccca gcgtggcgct ggacatgacc gcgcgcaaca tggaaccggg 16440
catgtatgcc tcaaaccggc cgtttatcaa ccgcctaatg gactacttgc atcgcgcggc 16500
cgccgtgaac cccgagtatt tcaccaatgc catcttgaac ccgcactggc taccgccccc 16560
tggtttctac accgggggat tcgaggtgcc cgagggtaac gatggattcc tctgggacga 16620
catagacgac agcgtgtttt ccccgcaacc gcagaccctg ctagagttgc aacagcgcga 16680
gcaggcagag gcggcgctgc gaaaggaaag cttccgcagg ccaagcagct tgtccgatct 16740
aggcgctgcg gccccgcggt cagatgctag tagcccattt ccaagcttga tagggtctct 16800
taccagcact cgcaccaccc gcccgcgcct gctgggcgag gaggagtacc taaacaactc 16860
gctgctgcag ccgcagcgcg aaaaaaacct gcctccggca tttcccaaca acgggataga 16920
gagcctagtg gacaagatga gtagatggaa gacgtacgcg caggagcaca gggacgtgcc 16980
aggcccgcgc ccgcccaccc gtcgtcaaag gcacgaccgt cagcggggtc tggtgtggga 17040
ggacgatgac tcggcagacg acagcagcgt cctggatttg ggagggagtg gcaacccgtt 17100
tgcgcacctt cgccccaggc tggggagaat gttttaaaaa aaaaaaagca tgatgcaaaa 17160
taaaaaactc accaaggcca tggcaccgag cgttggtttt cttgtattcc ccttagtatg 17220
cggcgcgcgg cgatgtatga ggaaggtcct cctccctcct acgagagtgt ggtgagcgcg 17280
gcgccagtgg cggcggcgct gggttctccc ttcgatgctc ccctggaccc gccgtttgtg 17340
cctccgcggt acctgcggcc taccgggggg agaaacagca tccgttactc tgagttggca 17400
cccctattcg acaccacccg tgtgtacctg gtggacaaca agtcaacgga tgtggcatcc 17460
ctgaactacc agaacgacca cagcaacttt ctgaccacgg tcattcaaaa caatgactac 17520
agcccggggg aggcaagcac acagaccatc aatcttgacg accggtcgca ctggggcggc 17580
gacctgaaaa ccatcctgca taccaacatg ccaaatgtga acgagttcat gtttaccaat 17640
aagtttaagg cgcgggtgat ggtgtcgcgc ttgcctacta aggacaatca ggtggagctg 17700
aaatacgagt gggtggagtt cacgctgccc gagggcaact actccgagac catgaccata 17760
gaccttatga acaacgcgat cgtggagcac tacttgaaag tgggcagaca gaacggggtt 17820
ctggaaagcg acatcggggt aaagtttgac acccgcaact tcagactggg gtttgacccc 17880
gtcactggtc ttgtcatgcc tggggtatat acaaacgaag ccttccatcc agacatcatt 17940
ttgctgccag gatgcggggt ggacttcacc cacagccgcc tgagcaactt gttgggcatc 18000
cgcaagcggc aacccttcca ggagggcttt aggatcacct acgatgatct ggagggtggt 18060
aacattcccg cactgttgga tgtggacgcc taccaggcga gcttgaaaga tgacaccgaa 18120
cagggcgggg gtggcgcagg cggcagcaac agcagtggca gcggcgcgga agagaactcc 18180
aacgcggcag ccgcggcaat gcagccggtg gaggacatga acgatcatgc cattcgcggc 18240
gacacctttg ccacacgggc tgaggagaag cgcgctgagg ccgaagcagc ggccgaagct 18300
gccgcccccg ctgcgcaacc cgaggtcgag aagcctcaga agaaaccggt gatcaaaccc 18360
ctgacagagg acagcaagaa acgcagttac aacctaataa gcaatgacag caccttcacc 18420
cagtaccgca gctggtacct tgcatacaac tacggcgacc ctcagaccgg aatccgctca 18480
tggaccctgc tttgcactcc tgacgtaacc tgcggctcgg agcaggtcta ctggtcgttg 18540
ccagacatga tgcaagaccc cgtgaccttc cgctccacgc gccagatcag caactttccg 18600
gtggtgggcg ccgagctgtt gcccgtgcac tccaagagct tctacaacga ccaggccgtc 18660
tactcccaac tcatccgcca gtttacctct ctgacccacg tgttcaatcg ctttcccgag 18720
aaccagattt tggcgcgccc gccagccccc accatcacca ccgtcagtga aaacgttcct 18780
gctctcacag atcacgggac gctaccgctg cgcaacagca tcggaggagt ccagcgagtg 18840
accattactg acgccagacg ccgcacctgc ccctacgttt acaaggccct gggcatagtc 18900
tcgccgcgcg tcctatcgag ccgcactttt tgagcaagca tgtccatcct tatatcgccc 18960
agcaataaca caggctgggg cctgcgcttc ccaagcaaga tgtttggcgg ggccaagaag 19020
cgctccgacc aacacccagt gcgcgtgcgc gggcactacc gcgcgccctg gggcgcgcac 19080
aaacgcggcc gcactgggcg caccaccgtc gatgacgcca tcgacgcggt ggtggaggag 19140
gcgcgcaact acacgcccac gccgccacca gtgtccacag tggacgcggc cattcagacc 19200
gtggtgcgcg gagcccggcg ctatgctaaa atgaagagac ggcggaggcg cgtagcacgt 19260
cgccaccgcc gccgacccgg cactgccgcc caacgcgcgg cggcggccct gcttaaccgc 19320
gcacgtcgca ccggccgacg ggcggccatg cgggccgctc gaaggctggc cgcgggtatt 19380
gtcactgtgc cccccaggtc caggcgacga gcggccgccg cagcagccgc ggccattagt 19440
gctatgactc agggtcgcag gggcaacgtg tattgggtgc gcgactcggt tagcggcctg 19500
cgcgtgcccg tgcgcacccg ccccccgcgc aactagattg caagaaaaaa ctacttagac 19560
tcgtactgtt gtatgtatcc agcggcggcg gcgcgcaacg aagctatgtc caagcgcaaa 19620
atcaaagaag agatgctcca ggtcatcgcg ccggagatct atggcccccc gaagaaggaa 19680
gagcaggatt acaagccccg aaagctaaag cgggtcaaaa agaaaaagaa agatgatgat 19740
gatgaacttg acgacgaggt ggaactgctg cacgctaccg cgcccaggcg acgggtacag 19800
tggaaaggtc gacgcgtaaa acgtgttttg cgacccggca ccaccgtagt ctttacgccc 19860
ggtgagcgct ccacccgcac ctacaagcgc gtgtatgatg aggtgtacgg cgacgaggac 19920
ctgcttgagc aggccaacga gcgcctcggg gagtttgcct acggaaagcg gcataaggac 19980
atgctggcgt tgccgctgga cgagggcaac ccaacaccta gcctaaagcc cgtaacactg 20040
cagcaggtgc tgcccgcgct tgcaccgtcc gaagaaaagc gcggcctaaa gcgcgagtct 20100
ggtgacttgg cacccaccgt gcagctgatg gtacccaagc gccagcgact ggaagatgtc 20160
ttggaaaaaa tgaccgtgga acctgggctg gagcccgagg tccgcgtgcg gccaatcaag 20220
caggtggcgc cgggactggg cgtgcagacc gtggacgttc agatacccac taccagtagc 20280
accagtattg ccaccgccac agagggcatg gagacacaaa cgtccccggt tgcctcagcg 20340
gtggcggatg ccgcggtgca ggcggtcgct gcggccgcgt ccaagacctc tacggaggtg 20400
caaacggacc cgtggatgtt tcgcgtttca gccccccggc gcccgcgcgg ttcgaggaag 20460
tacggcgccg ccagcgcgct actgcccgaa tatgccctac atccttccat tgcgcctacc 20520
cccggctatc gtggctacac ctaccgcccc agaagacgag caactacccg acgccgaacc 20580
accactggaa cccgccgccg ccgtcgccgt cgccagcccg tgctggcccc gatttccgtg 20640
cgcagggtgg ctcgcgaagg aggcaggacc ctggtgctgc caacagcgcg ctaccacccc 20700
agcatcgttt aaaagccggt ctttgtggtt cttgcagata tggccctcac ctgccgcctc 20760
cgtttcccgg tgccgggatt ccgaggaaga atgcaccgta ggaggggcat ggccggccac 20820
ggcctgacgg gcggcatgcg tcgtgcgcac caccggcggc ggcgcgcgtc gcaccgtcgc 20880
atgcgcggcg gtatcctgcc cctccttatt ccactgatcg ccgcggcgat tggcgccgtg 20940
cccggaattg catccgtggc cttgcaggcg cagagacact gattaaaaac aagttgcatg 21000
tggaaaaatc aaaataaaaa gtctggactc tcacgctcgc ttggtcctgt aactattttg 21060
tagaatggaa gacatcaact ttgcgtctct ggccccgcga cacggctcgc gcccgttcat 21120
gggaaactgg caagatatcg gcaccagcaa tatgagcggt ggcgccttca gctggggctc 21180
gctgtggagc ggcattaaaa atttcggttc caccgttaag aactatggca gcaaggcctg 21240
gaacagcagc acaggccaga tgctgaggga taagttgaaa gagcaaaatt tccaacaaaa 21300
ggtggtagat ggcctggcct ctggcattag cggggtggtg gacctggcca accaggcagt 21360
gcaaaataag attaacagta agcttgatcc ccgccctccc gtagaggagc ctccaccggc 21420
cgtggagaca gtgtctccag aggggcgtgg cgaaaagcgt ccgcgccccg acagggaaga 21480
aactctggtg acgcaaatag acgagcctcc ctcgtacgag gaggcactaa agcaaggcct 21540
gcccaccacc cgtcccatcg cgcccatggc taccggagtg ctgggccagc acacacccgt 21600
aacgctggac ctgcctcccc ccgccgacac ccagcagaaa cctgtgctgc caggcccgac 21660
cgccgttgtt gtaacccgtc ctagccgcgc gtccctgcgc cgcgccgcca gcggtccgcg 21720
atcgttgcgg cccgtagcca gtggcaactg gcaaagcaca ctgaacagca tcgtgggtct 21780
gggggtgcaa tccctgaagc gccgacgatg cttctgaata gctaacgtgt cgtatgtgtg 21840
tcatgtatgc gtccatgtcg ccgccagagg agctgctgag ccgccgcgcg cccgctttcc 21900
aagatggcta ccccttcgat gatgccgcag tggtcttaca tgcacatctc gggccaggac 21960
gcctcggagt acctgagccc cgggctggtg cagtttgccc gcgccaccga gacgtacttc 22020
agcctgaata acaagtttag aaaccccacg gtggcgccta cgcacgacgt gaccacagac 22080
cggtcccagc gtttgacgct gcggttcatc cctgtggacc gtgaggatac tgcgtactcg 22140
tacaaggcgc ggttcaccct agctgtgggt gataaccgtg tgctggacat ggcttccacg 22200
tactttgaca tccgcggcgt gctggacagg ggccctactt ttaagcccta ctctggcact 22260
gcctacaacg ccctggctcc caagggtgcc ccaaatcctt gcgaatggga tgaagctgct 22320
actgctcttg aaataaacct agaagaagag gacgatgaca acgaagacga agtagacgag 22380
caagctgagc agcaaaaaac tcacgtattt gggcaggcgc cttattctgg tataaatatt 22440
acaaaggagg gtattcaaat aggtgtcgaa ggtcaaacac ctaaatatgc cgataaaaca 22500
tttcaacctg aacctcaaat aggagaatct cagtggtacg aaactgaaat taatcatgca 22560
gctgggagag tccttaaaaa gactacccca atgaaaccat gttacggttc atatgcaaaa 22620
cccacaaatg aaaatggagg gcaaggcatt cttgtaaagc aacaaaatgg aaagctagaa 22680
agtcaagtgg aaatgcaatt tttctcaact actgaggcga ccgcaggcaa tggtgataac 22740
ttgactccta aagtggtatt gtacagtgaa gatgtagata tagaaacccc agacactcat 22800
atttcttaca tgcccactat taaggaaggt aactcacgag aactaatggg ccaacaatct 22860
atgcccaaca ggcctaatta cattgctttt agggacaatt ttattggtct aatgtattac 22920
aacagcacgg gtaatatggg tgttctggcg ggccaagcat cgcagttgaa tgctgttgta 22980
gatttgcaag acagaaacac agagctttca taccagcttt tgcttgattc cattggtgat 23040
agaaccaggt acttttctat gtggaatcag gctgttgaca gctatgatcc agatgttaga 23100
attattgaaa atcatggaac tgaagatgaa cttccaaatt actgctttcc actgggaggt 23160
gtgattaata cagagactct taccaaggta aaacctaaaa caggtcagga aaatggatgg 23220
gaaaaagatg ctacagaatt ttcagataaa aatgaaataa gagttggaaa taattttgcc 23280
atggaaatca atctaaatgc caacctgtgg agaaatttcc tgtactccaa catagcgctg 23340
tatttgcccg acaagctaaa gtacagtcct tccaacgtaa aaatttctga taacccaaac 23400
acctacgact acatgaacaa gcgagtggtg gctcccgggt tagtggactg ctacattaac 23460
cttggagcac gctggtccct tgactatatg gacaacgtca acccatttaa ccaccaccgc 23520
aatgctggcc tgcgctaccg ctcaatgttg ctgggcaatg gtcgctatgt gcccttccac 23580
atccaggtgc ctcagaagtt ctttgccatt aaaaacctcc ttctcctgcc gggctcatac 23640
acctacgagt ggaacttcag gaaggatgtt aacatggttc tgcagagctc cctaggaaat 23700
gacctaaggg ttgacggagc cagcattaag tttgatagca tttgccttta cgccaccttc 23760
ttccccatgg cccacaacac cgcctccacg cttgaggcca tgcttagaaa cgacaccaac 23820
gaccagtcct ttaacgacta tctctccgcc gccaacatgc tctaccctat acccgccaac 23880
gctaccaacg tgcccatatc catcccctcc cgcaactggg cggctttccg cggctgggcc 23940
ttcacgcgcc ttaagactaa ggaaacccca tcactgggct cgggctacga cccttattac 24000
acctactctg gctctatacc ctacctagat ggaacctttt acctcaacca cacctttaag 24060
aaggtggcca ttacctttga ctcttctgtc agctggcctg gcaatgaccg cctgcttacc 24120
cccaacgagt ttgaaattaa gcgctcagtt gacggggagg gttacaacgt tgcccagtgt 24180
aacatgacca aagactggtt cctggtacaa atgctagcta actacaacat tggctaccag 24240
ggcttctata tcccagagag ctacaaggac cgcatgtact ccttctttag aaacttccag 24300
cccatgagcc gtcaggtggt ggatgatact aaatacaagg actaccaaca ggtgggcatc 24360
ctacaccaac acaacaactc tggatttgtt ggctaccttg cccccaccat gcgcgaagga 24420
caggcctacc ctgctaactt cccctatccg cttataggca agaccgcagt tgacagcatt 24480
acccagaaaa agtttctttg cgatcgcacc ctttggcgca tcccattctc cagtaacttt 24540
atgtccatgg gcgcactcac agacctgggc caaaaccttc tctacgccaa ctccgcccac 24600
gcgctagaca tgacttttga ggtggatccc atggacgagc ccacccttct ttatgttttg 24660
tttgaagtct ttgacgtggt ccgtgtgcac cggccgcacc gcggcgtcat cgaaaccgtg 24720
tacctgcgca cgcccttctc ggccggcaac gccacaacat aaagaagcaa gcaacatcaa 24780
caacagctgc cgccatgggc tccagtgagc aggaactgaa agccattgtc aaagatcttg 24840
gttgtgggcc atattttttg ggcacctatg acaagcgctt tccaggcttt gtttctccac 24900
acaagctcgc ctgcgccata gtcaatacgg ccggtcgcga gactgggggc gtacactgga 24960
tggcctttgc ctggaacccg cactcaaaaa catgctacct ctttgagccc tttggctttt 25020
ctgaccagcg actcaagcag gtttaccagt ttgagtacga gtcactcctg cgccgtagcg 25080
ccattgcttc ttcccccgac cgctgtataa cgctggaaaa gtccacccaa agcgtacagg 25140
ggcccaactc ggccgcctgt ggactattct gctgcatgtt tctccacgcc tttgccaact 25200
ggccccaaac tcccatggat cacaacccca ccatgaacct tattaccggg gtacccaact 25260
ccatgctcaa cagtccccag gtacagccca ccctgcgtcg caaccaggaa cagctctaca 25320
gcttcctgga gcgccactcg ccctacttcc gcagccacag tgcgcagatt aggagcgcca 25380
cttctttttg tcacttgaaa aacatgtaaa aataatgtac tagagacact ttcaataaag 25440
gcaaatgctt ttatttgtac actctcgggt gattatttac ccccaccctt gccgtctgcg 25500
ccgtttaaaa atcaaagggg ttctgccgcg catcgctatg cgccactggc agggacacgt 25560
tgcgatactg gtgtttagtg ctccacttaa actcaggcac aaccatccgc ggcagctcgg 25620
tgaagttttc actccacagg ctgcgcacca tcaccaacgc gtttagcagg tcgggcgccg 25680
atatcttgaa gtcgcagttg gggcctccgc cctgcgcgcg cgagttgcga tacacagggt 25740
tgcagcactg gaacactatc agcgccgggt ggtgcacgct ggccagcacg ctcttgtcgg 25800
agatcagatc cgcgtccagg tcctccgcgt tgctcagggc gaacggagtc aactttggta 25860
gctgccttcc caaaaagggc gcgtgcccag gctttgagtt gcactcgcac cgtagtggca 25920
tcaaaaggtg accgtgcccg gtctgggcgt taggatacag cgcctgcata aaagccttga 25980
tctgcttaaa agccacctga gcctttgcgc cttcagagaa gaacatgccg caagacttgc 26040
cggaaaactg attggccgga caggccgcgt cgtgcacgca gcaccttgcg tcggtgttgg 26100
agatctgcac cacatttcgg ccccaccggt tcttcacgat cttggccttg ctagactgct 26160
ccttcagcgc gcgctgcccg ttttcgctcg tcacatccat ttcaatcacg tgctccttat 26220
ttatcataat gcttccgtgt agacacttaa gctcgccttc gatctcagcg cagcggtgca 26280
gccacaacgc gcagcccgtg ggctcgtgat gcttgtaggt cacctctgca aacgactgca 26340
ggtacgcctg caggaatcgc cccatcatcg tcacaaaggt cttgttgctg gtgaaggtca 26400
gctgcaaccc gcggtgctcc tcgttcagcc aggtcttgca tacggccgcc agagcttcca 26460
cttggtcagg cagtagtttg aagttcgcct ttagatcgtt atccacgtgg tacttgtcca 26520
tcagcgcgcg cgcagcctcc atgcccttct cccacgcaga cacgatcggc acactcagcg 26580
ggttcatcac cgtaatttca ctttccgctt cgctgggctc ttcctcttcc tcttgcgtcc 26640
gcataccacg cgccactggg tcgtcttcat tcagccgccg cactgtgcgc ttacctcctt 26700
tgccatgctt gattagcacc ggtgggttgc tgaaacccac catttgtagc gccacatctt 26760
ctctttcttc ctcgctgtcc acgattacct ctggtgatgg cgggcgctcg ggcttgggag 26820
aagggcgctt ctttttcttc ttgggcgcaa tggccaaatc cgccgccgag gtcgatggcc 26880
gcgggctggg tgtgcgcggc accagcgcgt cttgtgatga gtcttcctcg tcctcggact 26940
cgatacgccg cctcatccgc ttttttgggg gcgcccgggg aggcggcggc gacggggacg 27000
gggacgacac gtcctccatg gttgggggac gtcgcgccgc accgcgtccg cgctcggggg 27060
tggtttcgcg ctgctcctct tcccgactgg ccatttcctt ctcctatagg cagaaaaaga 27120
tcatggagtc agtcgagaag aaggacagcc taaccgcccc ctctgagttc gccaccaccg 27180
cctccaccga tgccgccaac gcgcctacca ccttccccgt cgaggcaccc ccgcttgagg 27240
aggaggaagt gattatcgag caggacccag gttttgtaag cgaagacgac gaggaccgct 27300
cagtaccaac agaggataaa aagcaagacc aggacaacgc agaggcaaac gaggaacaag 27360
tcgggcgggg ggacgaaagg catggcgact acctagatgt gggagacgac gtgctgttga 27420
agcatctgca gcgccagtgc gccattatct gcgacgcgtt gcaagagcgc agcgatgtgc 27480
ccctcgccat agcggatgtc agccttgcct acgaacgcca cctattctca ccgcgcgtac 27540
cccccaaacg ccaagaaaac ggcacatgcg agcccaaccc gcgcctcaac ttctaccccg 27600
tatttgccgt gccagaggtg cttgccacct atcacatctt tttccaaaac tgcaagatac 27660
ccctatcctg ccgtgccaac cgcagccgag cggacaagca gctggccttg cggcagggcg 27720
ctgtcatacc tgatatcgcc tcgctcaacg aagtgccaaa aatctttgag ggtcttggac 27780
gcgacgagaa gcgcgcggca aacgctctgc aacaggaaaa cagcgaaaat gaaagtcact 27840
ctggagtgtt ggtggaactc gagggtgaca acgcgcgcct agccgtacta aaacgcagca 27900
tcgaggtcac ccactttgcc tacccggcac ttaacctacc ccccaaggtc atgagcacag 27960
tcatgagtga gctgatcgtg cgccgtgcgc agcccctgga gagggatgca aatttgcaag 28020
aacaaacaga ggagggccta cccgcagttg gcgacgagca gctagcgcgc tggcttcaaa 28080
cgcgcgagcc tgccgacttg gaggagcgac gcaaactaat gatggccgca gtgctcgtta 28140
ccgtggagct tgagtgcatg cagcggttct ttgctgaccc ggagatgcag cgcaagctag 28200
aggaaacatt gcactacacc tttcgacagg gctacgtacg ccaggcctgc aagatctcca 28260
acgtggagct ctgcaacctg gtctcctacc ttggaatttt gcacgaaaac cgccttgggc 28320
aaaacgtgct tcattccacg ctcaagggcg aggcgcgccg cgactacgtc cgcgactgcg 28380
tttacttatt tctatgctac acctggcaga cggccatggg cgtttggcag cagtgcttgg 28440
aggagtgcaa cctcaaggag ctgcagaaac tgctaaagca aaacttgaag gacctatgga 28500
cggccttcaa cgagcgctcc gtggccgcgc acctggcgga catcattttc cccgaacgcc 28560
tgcttaaaac cctgcaacag ggtctgccag acttcaccag tcaaagcatg ttgcagaact 28620
ttaggaactt tatcctagag cgctcaggaa tcttgcccgc cacctgctgt gcacttccta 28680
gcgactttgt gcccattaag taccgcgaat gccctccgcc gctttggggc cactgctacc 28740
ttctgcagct agccaactac cttgcctacc actctgacat aatggaagac gtgagcggtg 28800
acggtctact ggagtgtcac tgtcgctgca acctatgcac cccgcaccgc tccctggttt 28860
gcaattcgca gctgcttaac gaaagtcaaa ttatcggtac ctttgagctg cagggtccct 28920
cgcctgacga aaagtccgcg gctccggggt tgaaactcac tccggggctg tggacgtcgg 28980
cttaccttcg caaatttgta cctgaggact accacgccca cgagattagg ttctacgaag 29040
accaatcccg cccgccaaat gcggagctta ccgcctgcgt cattacccag ggccacattc 29100
ttggccaatt gcaagccatc aacaaagccc gccaagagtt tctgctacga aagggacggg 29160
gggtttactt ggacccccag tccggcgagg agctcaaccc aatccccccg ccgccgcagc 29220
cctatcagca gcagccgcgg gcccttgctt cccaggatgg cacccaaaaa gaagctgcag 29280
ctgccgccgc cacccacgga cgaggaggaa tactgggaca gtcaggcaga ggaggttttg 29340
gacgaggagg aggaggacat gatggaagac tgggagagcc tagacgagga agcttccgag 29400
gtcgaagagg tgtcagacga aacaccgtca ccctcggtcg cattcccctc gccggcgccc 29460
cagaaatcgg caaccggttc cagcatggct acaacctccg ctcctcaggc gccgccggca 29520
ctgcccgttc gccgacccaa ccgtagatgg gacaccactg gaaccagggc cggtaagtcc 29580
aagcagccgc cgccgttagc ccaagagcaa caacagcgcc aaggctaccg ctcatggcgc 29640
gggcacaaga acgccatagt tgcttgcttg caagactgtg ggggcaacat ctccttcgcc 29700
cgccgctttc ttctctacca tcacggcgtg gccttccccc gtaacatcct gcattactac 29760
cgtcatctct acagcccata ctgcaccggc ggcagcggca gcggcagcaa cagcagcggc 29820
cacacagaag caaaggcgac cggatagcaa gactctgaca aagcccaaga aatccacagc 29880
ggcggcagca gcaggaggag gagcgctgcg tctggcgccc aacgaacccg tatcgacccg 29940
cgagcttaga aacaggattt ttcccactct gtatgctata tttcaacaga gcaggggcca 30000
agaacaagag ctgaaaataa aaaacaggtc tctgcgatcc ctcacccgca gctgcctgta 30060
tcacaaaagc gaagatcagc ttcggcgcac gctggaagac gcggaggctc tcttcagtaa 30120
atactgcgcg ctgactctta aggactagtt tcgcgccctt tctcaaattt aagcgcgaaa 30180
actacgtcat ctccagcggc cacacccggc gccagcacct gtcgtcagcg ccattatgag 30240
caaggaaatt cccacgccct acatgtggag ttaccagcca caaatgggac ttgcggctgg 30300
agctgcccaa gactactcaa cccgaataaa ctacatgagc gcgggacccc acatgatatc 30360
ccgggtcaac ggaatccgcg cccaccgaaa ccgaattctc ttggaacagg cggctattac 30420
caccacacct cgtaataacc ttaatccccg tagttggccc gctgccctgg tgtaccagga 30480
aagtcccgct cccaccactg tggtacttcc cagagacgcc caggccgaag ttcagatgac 30540
taactcaggg gcgcagcttg cgggcggctt tcgtcacagg gtgcggtcgc ccgggcaggg 30600
tataactcac ctgacaatca gagggcgagg tattcagctc aacgacgagt cggtgagctc 30660
ctcgcttggt ctccgtccgg acgggacatt tcagatcggc ggcgccggcc gtccttcatt 30720
cacgcctcgt caggcaatcc taactctgca gacctcgtcc tctgagccgc gctctggagg 30780
cattggaact ctgcaattta ttgaggagtt tgtgccatcg gtctacttta accccttctc 30840
gggacctccc ggccactatc cggatcaatt tattcctaac tttgacgcgg taaaggactc 30900
ggcggacggc tacgactgaa tgttaagtgg agaggcagag caactgcgcc tgaaacacct 30960
ggtccactgt cgccgccaca agtgctttgc ccgcgactcc ggtgagtttt gctactttga 31020
attgcccgag gatcatatcg agggcccggc gcacggcgtc cggcttaccg cccagggaga 31080
gcttgcccgt agcctgattc gggagtttac ccagcgcccc ctgctagttg agcgggacag 31140
gggaccctgt gttctcactg tgatttgcaa ctgtcctaac cttggattac atcaagatcc 31200
tctagttata actagagtac ccggggatct tattcccttt aactaataaa aaaaaataat 31260
aaagcatcac ttacttaaaa tcagttagca aatttctgtc cagtttattc agcagcacct 31320
ccttgccctc ctcccagctc tggtattgca gcttcctcct ggctgcaaac tttctccaca 31380
atctaaatgg aatgtcagtt tcctcctgtt cctgtccatc cgcacccact atcttcatgt 31440
tgttgcagat gaagcgcgca agaccgtctg aagatacctt caaccccgtg tatccatatg 31500
acacggaaac cggtcctcca actgtgcctt ttcttactcc tccctttgta tcccccaatg 31560
ggtttcaaga gagtccccct ggggtactct ctttgcgcct atccgaacct ctagttacct 31620
ccaatggcat gcttgcgctc aaaatgggca acggcctctc tctggacgag gccggcaacc 31680
ttacctccca aaatgtaacc actgtgagcc cacctctcaa aaaaaccaag tcaaacataa 31740
acctggaaat atctgcaccc ctcacagtta cctcagaagc cctaactgtg gctgccgccg 31800
cacctctaat ggtcgcgggc aacacactca ccatgcaatc acaggccccg ctaaccgtgc 31860
acgactccaa acttagcatt gccacccaag gacccctcac agtgtcagaa ggaaagctag 31920
ccctgcaaac atcaggcccc ctcaccacca ccgatagcag tacccttact atcactgcct 31980
caccccctct aactactgcc actggtagct tgggcattga cttgaaagag cccatttata 32040
cacaaaatgg aaaactagga ctaaagtacg gggctccttt gcatgtaaca gacgacctaa 32100
acactttgac cgtagcaact ggtccaggtg tgactattaa taatacttcc ttgcaaacta 32160
aagttactgg agccttgggt tttgattcac aaggcaatat gcaacttaat gtagcaggag 32220
gactaaggat tgattctcaa aacagacgcc ttatacttga tgttagttat ccgtttgatg 32280
ctcaaaacca actaaatcta agactaggac agggccctct ttttataaac tcagcccaca 32340
acttggatat taactacaac aaaggccttt acttgtttac agcttcaaac aattccaaaa 32400
agcttgaggt taacctaagc actgccaagg ggttgatgtt tgacgctaca gccatagcca 32460
ttaatgcagg agatgggctt gaatttggtt cacctaatgc accaaacaca aatcccctca 32520
aaacaaaaat tggccatggc ctagaatttg attcaaacaa ggctatggtt cctaaactag 32580
gaactggcct tagttttgac agcacaggtg ccattacagt aggaaacaaa aataatgata 32640
agctaacttt gtggaccaca ccagctccat ctcctaactg tagactaaat gcagagaaag 32700
atgctaaact cactttggtc ttaacaaaat gtggcagtca aatacttgct acagtttcag 32760
ttttggctgt taaaggcagt ttggctccaa tatctggaac agttcaaagt gctcatctta 32820
ttataagatt tgacgaaaat ggagtgctac taaacaattc cttcctggac ccagaatatt 32880
ggaactttag aaatggagat cttactgaag gcacagccta tacaaacgct gttggattta 32940
tgcctaacct atcagcttat ccaaaatctc acggtaaaac tgccaaaagt aacattgtca 33000
gtcaagttta cttaaacgga gacaaaacta aacctgtaac actaaccatt acactaaacg 33060
gtacacagga aacaggagac acaactccaa gtgcatactc tatgtcattt tcatgggact 33120
ggtctggcca caactacatt aatgaaatat ttgccacatc ctcttacact ttttcataca 33180
ttgcccaaga ataaagaatc gtttgtgtta tgtttcaacg tgtttatttt tcaattgcag 33240
aaaatttcaa gtcatttttc attcagtagt atagccccac caccacatag cttatacaga 33300
tcaccgtacc ttaatcaaac tcacagaacc ctagtattca acctgccacc tccctcccaa 33360
cacacagagt acacagtcct ttctccccgg ctggccttaa aaagcatcat atcatgggta 33420
acagacatat tcttaggtgt tatattccac acggtttcct gtcgagccaa acgctcatca 33480
gtgatattaa taaactcccc gggcagctca cttaagttca tgtcgctgtc cagctgctga 33540
gccacaggct gctgtccaac ttgcggttgc ttaacgggcg gcgaaggaga agtccacgcc 33600
tacatggggg tagagtcata atcgtgcatc aggatagggc ggtggtgctg cagcagcgcg 33660
cgaataaact gctgccgccg ccgctccgtc ctgcaggaat acaacatggc agtggtctcc 33720
tcagcgatga ttcgcaccgc ccgcagcata aggcgccttg tcctccgggc acagcagcgc 33780
accctgatct cacttaaatc agcacagtaa ctgcagcaca gcaccacaat attgttcaaa 33840
atcccacagt gcaaggcgct gtatccaaag ctcatggcgg ggaccacaga acccacgtgg 33900
ccatcatacc acaagcgcag gtagattaag tggcgacccc tcataaacac gctggacata 33960
aacattacct cttttggcat gttgtaattc accacctccc ggtaccatat aaacctctga 34020
ttaaacatgg cgccatccac caccatccta aaccagctgg ccaaaacctg cccgccggct 34080
atacactgca gggaaccggg actggaacaa tgacagtgga gagcccagga ctcgtaacca 34140
tggatcatca tgctcgtcat gatatcaatg ttggcacaac acaggcacac gtgcatacac 34200
ttcctcagga ttacaagctc ctcccgcgtt agaaccatat cccagggaac aacccattcc 34260
tgaatcagcg taaatcccac actgcaggga agacctcgca cgtaactcac gttgtgcatt 34320
gtcaaagtgt tacattcggg cagcagcgga tgatcctcca gtatggtagc gcgggtttct 34380
gtctcaaaag gaggtagacg atccctactg tacggagtgc gccgagacaa ccgagatcgt 34440
gttggtcgta gtgtcatgcc aaatggaacg ccggacgtag tcatatttcc tgaagcaaaa 34500
ccaggtgcgg gcgtgacaaa cagatctgcg tctccggtct cgccgcttag atcgctctgt 34560
gtagtagttg tagtatatcc actctctcaa agcatccagg cgccccctgg cttcgggttc 34620
tatgtaaact ccttcatgcg ccgctgccct gataacatcc accaccgcag aataagccac 34680
acccagccaa cctacacatt cgttctgcga gtcacacacg ggaggagcgg gaagagctgg 34740
aagaaccatg tttttttttt tattccaaaa gattatccaa aacctcaaaa tgaagatcta 34800
ttaagtgaac gcgctcccct ccggtggcgt ggtcaaactc tacagccaaa gaacagataa 34860
tggcatttgt aagatgttgc acaatggctt ccaaaaggca aacggccctc acgtccaagt 34920
ggacgtaaag gctaaaccct tcagggtgaa tctcctctat aaacattcca gcaccttcaa 34980
ccatgcccaa ataattctca tctcgccacc ttctcaatat atctctaagc aaatcccgaa 35040
tattaagtcc ggccattgta aaaatctgct ccagagcgcc ctccaccttc agcctcaagc 35100
agcgaatcat gattgcaaaa attcaggttc ctcacagacc tgtataagat tcaaaagcgg 35160
aacattaaca aaaataccgc gatcccgtag gtcccttcgc agggccagct gaacataatc 35220
gtgcaggtct gcacggacca gcgcggccac ttccccgcca ggaaccttga caaaagaacc 35280
cacactgatt atgacacgca tactcggagc tatgctaacc agcgtagccc cgatgtaagc 35340
tttgttgcat gggcggcgat ataaaatgca aggtgctgct caaaaaatca ggcaaagcct 35400
cgcgcaaaaa agaaagcaca tcgtagtcat gctcatgcag ataaaggcag gtaagctccg 35460
gaaccaccac agaaaaagac accatttttc tctcaaacat gtctgcgggt ttctgcataa 35520
acacaaaata aaataacaaa aaaacattta aacattagaa gcctgtctta caacaggaaa 35580
aacaaccctt ataagcataa gacggactac ggccatgccg gcgtgaccgt aaaaaaactg 35640
gtcaccgtga ttaaaaagca ccaccgacag ctcctcggtc atgtccggag tcataatgta 35700
agactcggta aacacatcag gttgattcat cggtcagtgc taaaaagcga ccgaaatagc 35760
ccgggggaat acatacccgc aggcgtagag acaacattac agcccccata ggaggtataa 35820
caaaattaat aggagagaaa aacacataaa cacctgaaaa accctcctgc ctaggcaaaa 35880
tagcaccctc ccgctccaga acaacataca gcgcttcaca gcggcagcct aacagtcagc 35940
cttaccagta aaaaagaaaa cctattaaaa aaacaccact cgacacggca ccagctcaat 36000
cagtcacagt gtaaaaaagg gccaagtgca gagcgagtat atataggact aaaaaatgac 36060
gtaacggtta aagtccacaa aaaacaccca gaaaaccgca cgcgaaccta cgcccagaaa 36120
cgaaagccaa aaaacccaca acttcctcaa atcgtcactt ccgttttccc acgttacgta 36180
acttcccatt ttaagaaaac tacaattccc aacacataca agttactccg ccctaaaacc 36240
tacgtcaccc gccccgttcc cacgccccgc gccacgtcac aaactccacc ccctcattat 36300
catattggct tcaatccaaa ataaggtata ttattgatga tact 36344
<211> 37934
<212> DNA
<213> Artificial(人工序列)
ctttaattaa ggatcccgac ctgtcctcga ccgatgccct tgagagcctt caacccagtc 60
agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt 120
atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc 180
tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc 240
ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt 300
atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc 360
tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg 420
caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc 480
gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc 540
gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc 600
tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc 660
ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga 720
gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca 780
gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg 840
tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg 900
aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag 960
caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag 1020
cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa 1080
cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc 1140
gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat 1200
catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga 1260
acagaaattc ccccttacac ggaggcatca agtgaccaaa caggaaaaaa ccgcccttaa 1320
catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga 1380
cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg 1440
cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga 1500
gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc 1560
agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt 1620
gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 1680
tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 1740
tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 1800
aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 1860
aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 1920
ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 1980
acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2040
ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2100
tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 2160
tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 2220
gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 2280
agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 2340
tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 2400
agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 2460
tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 2520
acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 2580
tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 2640
agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 2700
tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 2760
acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 2820
tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 2880
ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 2940
agtagttcgc cagttaatag tttgcgcaac gttgttgtca aaaaggatct tcacctagat 3000
ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 3060
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 3120
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 3180
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctc 3240
gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag gatgaggatc 3300
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 3360
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 3420
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 3480
tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 3540
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 3600
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 3660
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 3720
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 3780
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat 3840
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 3900
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 3960
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 4020
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 4080
ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc tcatttttta 4140
accaataggc cgaaatcggc aacatccctt ataaatcaaa agaatagacc gcgatagggt 4200
tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 4260
aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa 4320
gttttttgcg gtcgaggtgc cgtaaagctc taaatcggaa ccctaaaggg agcccccgat 4380
ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 4440
gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 4500
cgcgcttaat gcgccggatt aattaatcag tcccttccag ctctctgccc cttttggatt 4560
gaagccaata tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg 4620
gcgggtgacg tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa 4680
gcgacggatg tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa 4740
ttttcgcgcg gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg 4800
ccattttcgc gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca 4860
tagcgcgtaa tcgcgcgtta agatacattg atgagtttgg acaaaccaca actagaatgc 4920
agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 4980
taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 5040
gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt 5100
atgatcagtt atctagatcc ggtggatctg agtccggact tgtacagctc gtccatgccg 5160
agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg 5220
gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg 5280
ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg 5340
atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata 5400
tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc 5460
ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc 5520
tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag 5580
ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg 5640
aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg 5700
ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg 5760
ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc 5820
accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcgga 5880
tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc 5940
ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt 6000
gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa 6060
accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga 6120
ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat 6180
gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca 6240
cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga 6300
aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt 6360
cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat 6420
atgggctatg aactaatgac cccgtaattg attactatta gtcctagcag atcctggttc 6480
tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc 6540
aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca 6600
gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca 6660
gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag 6720
tcgaggctga tcagcgggtt tagcggccga tatcaagcgc tcacgggata ctcgatttaa 6780
attgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga 6840
ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca 6900
aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg 6960
tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg 7020
gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta 7080
aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata 7140
ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag 7200
ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga 7260
tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca 7320
gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt 7380
caggctccct tatacacagc cagtctgcag ctcgctcttc atttaaattc cgcgtccgct 7440
ttccgattcc tgaattcggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca 7500
ggcgatctga cggttcacta aacagctcga tctctatcac tgatagggag atctctatca 7560
ctgataggga gatctctatc actgataggg agatctctat cactgatagg gagagctctg 7620
cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg 7680
ttacgacatt ttggaaagtc ccgttgattt tggttccaaa acaaactccc attgacgtca 7740
atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac 7800
tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta 7860
ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg 7920
acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt 7980
taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga 8040
acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca 8100
tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta 8160
attgattact attaataact agagatcgta agggtgggaa agaatatata aggtgggggt 8220
cttatgtagt tttgtatctg ttttgcagca gccgccgccg ccatgagcac caactcgttt 8280
gatggaagca ttgtgagctc atatttgaca acgcgcatgc ccccatgggc cggggtgcgt 8340
cagaatgtga tgggctccag cattgatggt cgccccgtcc tgcccgcaaa ctctactacc 8400
ttgacctacg agaccgtgtc tggaacgccg ttggagactg cagcctccgc cgccgcttca 8460
gccgctgcag ccaccgcccg cgggattgtg actgactttg ctttcctgag cccgcttgca 8520
agcagtgcag cttcccgttc atccgcccgc gatgacaagt tgacggctct tttggcacaa 8580
ttggattctt tgacccggga acttaatgtc gtttctcagc agctgttgga tctgcgccag 8640
caggtttctg ccctgaaggc ttcctcccct cccaatgcgg tttaaaacat aaataaaaaa 8700
ccagactctg tttggatttg gatcaagcaa gtgtcttgct gtctttattt aggggttttg 8760
cgcgcgcggt aggcccggga ccagcggtct cggtcgttga gggtcctgtg tattttttcc 8820
aggacgtggt aaaggtgact ctggatgttc agatacatgg gcataagccc gtctctgggg 8880
tggaggtagc accactgcag agcttcatgc tgcggggtgg tgttgtagat gatccagtcg 8940
tagcaggagc gctgggcgtg gtgcctaaaa atgtctttca gtagcaagct gattgccagg 9000
ggcaggccct tggtgtaagt gtttacaaag cggttaagct gggatgggtg catacgtggg 9060
gatatgagat gcatcttgga ctgtattttt aggttggcta tgttcccagc catatccctc 9120
cggggattca tgttgtgcag aaccaccagc acagtgtatc cggtgcactt gggaaatttg 9180
tcatgtagct tagaaggaaa tgcgtggaag aacttggaga cgcccttgtg acctccaaga 9240
ttttccatgc attcgtccat aatgatggca atgggcccac gggcggcggc ctgggcgaag 9300
atatttctgg gatcactaac gtcatagttg tgttccagga tgagatcgtc ataggccatt 9360
tttacaaagc gcgggcggag ggtgccagac tgcggtataa tggttccatc cggcccaggg 9420
gcgtagttac cctcacagat ttgcatttcc cacgctttga gttcagatgg ggggatcatg 9480
tctacctgcg gggcgatgaa gaaaacggtt tccggggtag gggagatcag ctgggaagaa 9540
agcaggttcc tgagcagctg cgacttaccg cagccggtgg gcccgtaaat cacacctatt 9600
accgggtgca actggtagtt aagagagctg cagctgccgt catccctgag caggggggcc 9660
acttcgttaa gcatgtccct gactcgcatg ttttccctga ccaaatccgc cagaaggcgc 9720
tcgccgccca gcgatagcag ttcttgcaag gaagcaaagt ttttcaacgg tttgagaccg 9780
tccgccgtag gcatgctttt gagcgtttga ccaagcagtt ccaggcggtc ccacagctcg 9840
gtcacctgct ctacggcatc tcgatccagc atatctcctc gtttcgcggg ttggggcggc 9900
tttcgctgta cggcagtagt cggtgctcgt ccagacgggc cagggtcatg tctttccacg 9960
ggcgcagggt cctcgtcagc gtagtctggg tcacggtgaa ggggtgcgct ccgggctgcg 10020
cgctggccag ggtgcgcttg aggctggtcc tgctggtgct gaagcgctgc cggtcttcgc 10080
cctgcgcgtc ggccaggtag catttgacca tggtgtcata gtccagcccc tccgcggcgt 10140
ggcccttggc gcgcagcttg cccttggagg aggcgccgca cgaggggcag tgcagacttt 10200
tgagggcgta gagcttgggc gcgagaaata ccgattccgg ggagtaggca tccgcgccgc 10260
aggccccgca gacggtctcg cattccacga gccaggtgag ctctggccgt tcggggtcaa 10320
aaaccaggtt tcccccatgc tttttgatgc gtttcttacc tctggtttcc atgagccggt 10380
gtccacgctc ggtgacgaaa aggctgtccg tgtccccgta tacagacttg agaggcctgt 10440
cctcgagcgg tgttccgcgg tcctcctcgt atagaaactc ggaccactct gagacaaagg 10500
ctcgcgtcca ggccagcacg aaggaggcta agtgggaggg gtagcggtcg ttgtccacta 10560
gggggtccac tcgctccagg gtgtgaagac acatgtcgcc ctcttcggca tcaaggaagg 10620
tgattggttt gtaggtgtag gccacgtgac cgggtgttcc tgaagggggg ctataaaagg 10680
gggtgggggc gcgttcgtcc tcactctctt ccgcatcgct gtctgcgagg gccagctgtt 10740
ggggtgagta ctccctctga aaagcgggca tgacttctgc gctaagattg tcagtttcca 10800
aaaacgagga ggatttgata ttcacctggc ccgcggtgat gcctttgagg gtggccgcat 10860
ccatctggtc agaaaagaca atctttttgt tgtcaagctt ggtggcaaac gacccgtaga 10920
gggcgttgga cagcaacttg gcgatggagc gcagggtttg gtttttgtcg cgatcggcgc 10980
gctccttggc cgcgatgttt agctgcacgt attcgcgcgc aacgcaccgc cattcgggaa 11040
agacggtggt gcgctcgtcg ggcaccaggt gcacgcgcca accgcggttg tgcagggtga 11100
caaggtcaac gctggtggct acctctccgc gtaggcgctc gttggtccag cagaggcggc 11160
cgcccttgcg cgagcagaat ggcggtaggg ggtctagctg cgtctcgtcc ggggggtctg 11220
cgtccacggt aaagaccccg ggcagcaggc gcgcgtcgaa gtagtctatc ttgcatcctt 11280
gcaagtctag cgcctgctgc catgcgcggg cggcaagcgc gcgctcgtat gggttgagtg 11340
ggggacccca tggcatgggg tgggtgagcg cggaggcgta catgccgcaa atgtcgtaaa 11400
cgtagagggg ctctctgagt attccaagat atgtagggta gcatcttcca ccgcggatgc 11460
tggcgcgcac gtaatcgtat agttcgtgcg agggagcgag gaggtcggga ccgaggttgc 11520
tacgggcggg ctgctctgct cggaagacta tctgcctgaa gatggcatgt gagttggatg 11580
atatggttgg acgctggaag acgttgaagc tggcgtctgt gagacctacc gcgtcacgca 11640
cgaaggaggc gtaggagtcg cgcagcttgt tgaccagctc ggcggtgacc tgcacgtcta 11700
gggcgcagta gtccagggtt tccttgatga tgtcatactt atcctgtccc ttttttttcc 11760
acagctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc 11820
cgtcggcctc cgaacggtaa gagcctagca tgtagaactg gttgacggcc tggtaggcgc 11880
agcatccctt ttctacgggt agcgcgtatg cctgcgcggc cttccggagc gaggtgtggg 11940
tgagcgcaaa ggtgtccctg accatgactt tgaggtactg gtatttgaag tcagtgtcgt 12000
cgcatccgcc ctgctcccag agcaaaaagt ccgtgcgctt tttggaacgc ggatttggca 12060
gggcgaaggt gacatcgttg aagagtatct ttcccgcgcg aggcataaag ttgcgtgtga 12120
tgcggaaggg tcccggcacc tcggaacggt tgttaattac ctgggcggcg agcacgatct 12180
cgtcaaagcc gttgatgttg tggcccacaa tgtaaagttc caagaagcgc gggatgccct 12240
tgatggaagg caatttttta agttcctcgt aggtgagctc ttcaggggag ctgagcccgt 12300
gctctgaaag ggcccagtct gcaagatgag ggttggaagc gacgaatgag ctccacaggt 12360
cacgggccat tagcatttgc aggtggtcgc gaaaggtcct aaactggcga cctatggcca 12420
ttttttctgg ggtgatgcag tagaaggtaa gcgggtcttg ttcccagcgg tcccatccaa 12480
ggttcgcggc taggtctcgc gcggcagtca ctagaggctc atctccgccg aacttcatga 12540
ccagcatgaa gggcacgagc tgcttcccaa aggcccccat ccaagtatag gtctctacat 12600
cgtaggtgac aaagagacgc tcggtgcgag gatgcgagcc gatcgggaag aactggatct 12660
cccgccacca attggaggag tggctattga tgtggtgaaa gtagaagtcc ctgcgacggg 12720
ccgaacactc gtgctggctt ttgtaaaaac gtgcgcagta ctggcagcgg tgcacgggct 12780
gtacatcctg cacgaggttg acctgacgac cgcgcacaag gaagcagagt gggaatttga 12840
gcccctcgcc tggcgggttt ggctggtggt cttctacttc ggctgcttgt ccttgaccgt 12900
ctggctgctc gaggggagtt acggtggatc ggaccaccac gccgcgcgag cccaaagtcc 12960
agatgtccgc gcgcggcggt cggagcttga tgacaacatc gcgcagatgg gagctgtcca 13020
tggtctggag ctcccgcggc gtcaggtcag gcgggagctc ctgcaggttt acctcgcata 13080
gacgggtcag ggcgcgggct agatccaggt gatacctaat ttccaggggc tggttggtgg 13140
cggcgtcgat ggcttgcaag aggccgcatc cccgcggcgc gactacggta ccgcgcggcg 13200
ggcggtgggc cgcgggggtg tccttggatg atgcatctaa aagcggtgac gcgggcgagc 13260
ccccggaggt agggggggct ccggacccgc cgggagaggg ggcaggggca cgtcggcgcc 13320
gcgcgcgggc aggagctggt gctgcgcgcg taggttgctg gcgaacgcga cgacgcggcg 13380
gttgatctcc tgaatctggc gcctctgcgt gaagacgacg ggcccggtga gcttgagcct 13440
gaaagagagt tcgacagaat caatttcggt gtcgttgacg gcggcctggc gcaaaatctc 13500
ctgcacgtct cctgagttgt cttgataggc gatctcggcc atgaactgct cgatctcttc 13560
ctcctggaga tctccgcgtc cggctcgctc cacggtggcg gcgaggtcgt tggaaatgcg 13620
ggccatgagc tgcgagaagg cgttgaggcc tccctcgttc cagacgcggc tgtagaccac 13680
gcccccttcg gcatcgcggg cgcgcatgac cacctgcgcg agattgagct ccacgtgccg 13740
ggcgaagacg gcgtagtttc gcaggcgctg aaagaggtag ttgagggtgg tggcggtgtg 13800
ttctgccacg aagaagtaca taacccagcg tcgcaacgtg gattcgttga tatcccccaa 13860
ggcctcaagg cgctccatgg cctcgtagaa gtccacggcg aagttgaaaa actgggagtt 13920
gcgcgccgac acggttaact cctcctccag aagacggatg agctcggcga cagtgtcgcg 13980
cacctcgcgc tcaaaggcta caggggcctc ttcttcttct tcaatctcct cttccataag 14040
ggcctcccct tcttcttctt ctggcggcgg tgggggaggg gggacacggc ggcgacgacg 14100
gcgcaccggg aggcggtcga caaagcgctc gatcatctcc ccgcggcgac ggcgcatggt 14160
ctcggtgacg gcgcggccgt tctcgcgggg gcgcagttgg aagacgccgc ccgtcatgtc 14220
ccggttatgg gttggcgggg ggctgccatg cggcagggat acggcgctaa cgatgcatct 14280
caacaattgt tgtgtaggta ctccgccgcc gagggacctg agcgagtccg catcgaccgg 14340
atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac 14400
cgtggcgggc ggcagcgggc ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat 14460
gtaattaaag taggcggtct tgagacggcg gatggtcgac agaagcacca tgtccttggg 14520
tccggcctgc tgaatgcgca ggcggtcggc catgccccag gcttcgtttt gacatcggcg 14580
caggtctttg tagtagtctt gcatgagcct ttctaccggc acttcttctt ctccttcctc 14640
ttgtcctgca tctcttgcat ctatcgctgc ggcggcggcg gagtttggcc gtaggtggcg 14700
ccctcttcct cccatgcgtg tgaccccgaa gcccctcatc ggctgaagca gggctaggtc 14760
ggcgacaacg cgctcggcta atatggcctg ctgcacctgc gtgagggtag actggaagtc 14820
atccatgtcc acaaagcggt ggtatgcgcc cgtgttgatg gtgtaagtgc agttggccat 14880
aacggaccag ttaacggtct ggtgacccgg ctgcgagagc tcggtgtacc tgagacgcga 14940
gtaagccctc gagtcaaata cgtagtcgtt gcaagtccgc accaggtact ggtatcccac 15000
caaaaagtgc ggcggcggct ggcggtagag gggccagcgt agggtggccg gggctccggg 15060
ggcgagatct tccaacataa ggcgatgata tccgtagatg tacctggaca tccaggtgat 15120
gccggcggcg gtggtggagg cgcgcggaaa gtcgcggacg cggttccaga tgttgcgcag 15180
cggcaaaaag tgctccatgg tcgggacgct ctggccggtc aggcgcgcgc aatcgttgac 15240
gctctaccgt gcaaaaggag agcctgtaag cgggcactct tccgtggtct ggtggataaa 15300
ttcgcaaggg tatcatggcg gacgaccggg gttcgagccc cgtatccggc cgtccgccgt 15360
gatccatgcg gttaccgccc gcgtgtcgaa cccaggtgtg cgacgtcaga caacggggga 15420
gtgctccttt tggcttcctt ccaggcgcgg cggctgctgc gctagctttt ttggccactg 15480
gccgcgcgca gcgtaagcgg ttaggctgga aagcgaaagc attaagtggc tcgctccctg 15540
tagccggagg gttattttcc aagggttgag tcgcgggacc cccggttcga gtctcggacc 15600
ggccggactg cggcgaacgg gggtttgcct ccccgtcatg caagaccccg cttgcaaatt 15660
cctccggaaa cagggacgag cccctttttt gcttttccca gatgcatccg gtgctgcggc 15720
agatgcgccc ccctcctcag cagcggcaag agcaagagca gcggcagaca tgcagggcac 15780
cctcccctcc tcctaccgcg tcaggagggg cgacatccgc ggttgacgcg gcagcagatg 15840
gtgattacga acccccgcgg cgccgggccc ggcactacct ggacttggag gagggcgagg 15900
gcctggcgcg gctaggagcg ccctctcctg agcggtaccc aagggtgcag ctgaagcgtg 15960
atacgcgtga ggcgtacgtg ccgcggcaga acctgtttcg cgaccgcgag ggagaggagc 16020
ccgaggagat gcgggatcga aagttccacg cagggcgcga gctgcggcat ggcctgaatc 16080
gcgagcggtt gctgcgcgag gaggactttg agcccgacgc gcgaaccggg attagtcccg 16140
cgcgcgcaca cgtggcggcc gccgacctgg taaccgcata cgagcagacg gtgaaccagg 16200
agattaactt tcaaaaaagc tttaacaacc acgtgcgtac gcttgtggcg cgcgaggagg 16260
tggctatagg actgatgcat ctgtgggact ttgtaagcgc gctggagcaa aacccaaata 16320
gcaagccgct catggcgcag ctgttcctta tagtgcagca cagcagggac aacgaggcat 16380
tcagggatgc gctgctaaac atagtagagc ccgagggccg ctggctgctc gatttgataa 16440
acatcctgca gagcatagtg gtgcaggagc gcagcttgag cctggctgac aaggtggccg 16500
ccatcaacta ttccatgctt agcctgggca agttttacgc ccgcaagata taccataccc 16560
cttacgttcc catagacaag gaggtaaaga tcgaggggtt ctacatgcgc atggcgctga 16620
aggtgcttac cttgagcgac gacctgggcg tttatcgcaa cgagcgcatc cacaaggccg 16680
tgagcgtgag ccggcggcgc gagctcagcg accgcgagct gatgcacagc ctgcaaaggg 16740
ccctggctgg cacgggcagc ggcgatagag aggccgagtc ctactttgac gcgggcgctg 16800
acctgcgctg ggccccaagc cgacgcgccc tggaggcagc tggggccgga cctgggctgg 16860
cggtggcacc cgcgcgcgct ggcaacgtcg gcggcgtgga ggaatatgac gaggacgatg 16920
agtacgagcc agaggacggc gagtactaag cggtgatgtt tctgatcaga tgatgcaaga 16980
cgcaacggac ccggcggtgc gggcggcgct gcagagccag ccgtccggcc ttaactccac 17040
ggacgactgg cgccaggtca tggaccgcat catgtcgctg actgcgcgca atcctgacgc 17100
gttccggcag cagccgcagg ccaaccggct ctccgcaatt ctggaagcgg tggtcccggc 17160
gcgcgcaaac cccacgcacg agaaggtgct ggcgatcgta aacgcgctgg ccgaaaacag 17220
ggccatccgg cccgacgagg ccggcctggt ctacgacgcg ctgcttcagc gcgtggctcg 17280
ttacaacagc ggcaacgtgc agaccaacct ggaccggctg gtgggggatg tgcgcgaggc 17340
cgtggcgcag cgtgagcgcg cgcagcagca gggcaacctg ggctccatgg ttgcactaaa 17400
cgccttcctg agtacacagc ccgccaacgt gccgcgggga caggaggact acaccaactt 17460
tgtgagcgca ctgcggctaa tggtgactga gacaccgcaa agtgaggtgt accagtctgg 17520
gccagactat tttttccaga ccagtagaca aggcctgcag accgtaaacc tgagccaggc 17580
tttcaaaaac ttgcaggggc tgtggggggt gcgggctccc acaggcgacc gcgcgaccgt 17640
gtctagcttg ctgacgccca actcgcgcct gttgctgctg ctaatagcgc ccttcacgga 17700
cagtggcagc gtgtcccggg acacatacct aggtcacttg ctgacactgt accgcgaggc 17760
cataggtcag gcgcatgtgg acgagcatac tttccaggag attacaagtg tcagccgcgc 17820
gctggggcag gaggacacgg gcagcctgga ggcaacccta aactacctgc tgaccaaccg 17880
gcggcagaag atcccctcgt tgcacagttt aaacagcgag gaggagcgca ttttgcgcta 17940
cgtgcagcag agcgtgagcc ttaacctgat gcgcgacggg gtaacgccca gcgtggcgct 18000
ggacatgacc gcgcgcaaca tggaaccggg catgtatgcc tcaaaccggc cgtttatcaa 18060
ccgcctaatg gactacttgc atcgcgcggc cgccgtgaac cccgagtatt tcaccaatgc 18120
catcttgaac ccgcactggc taccgccccc tggtttctac accgggggat tcgaggtgcc 18180
cgagggtaac gatggattcc tctgggacga catagacgac agcgtgtttt ccccgcaacc 18240
gcagaccctg ctagagttgc aacagcgcga gcaggcagag gcggcgctgc gaaaggaaag 18300
cttccgcagg ccaagcagct tgtccgatct aggcgctgcg gccccgcggt cagatgctag 18360
tagcccattt ccaagcttga tagggtctct taccagcact cgcaccaccc gcccgcgcct 18420
gctgggcgag gaggagtacc taaacaactc gctgctgcag ccgcagcgcg aaaaaaacct 18480
gcctccggca tttcccaaca acgggataga gagcctagtg gacaagatga gtagatggaa 18540
gacgtacgcg caggagcaca gggacgtgcc aggcccgcgc ccgcccaccc gtcgtcaaag 18600
gcacgaccgt cagcggggtc tggtgtggga ggacgatgac tcggcagacg acagcagcgt 18660
cctggatttg ggagggagtg gcaacccgtt tgcgcacctt cgccccaggc tggggagaat 18720
gttttaaaaa aaaaaaagca tgatgcaaaa taaaaaactc accaaggcca tggcaccgag 18780
cgttggtttt cttgtattcc ccttagtatg cggcgcgcgg cgatgtatga ggaaggtcct 18840
cctccctcct acgagagtgt ggtgagcgcg gcgccagtgg cggcggcgct gggttctccc 18900
ttcgatgctc ccctggaccc gccgtttgtg cctccgcggt acctgcggcc taccgggggg 18960
agaaacagca tccgttactc tgagttggca cccctattcg acaccacccg tgtgtacctg 19020
gtggacaaca agtcaacgga tgtggcatcc ctgaactacc agaacgacca cagcaacttt 19080
ctgaccacgg tcattcaaaa caatgactac agcccggggg aggcaagcac acagaccatc 19140
aatcttgacg accggtcgca ctggggcggc gacctgaaaa ccatcctgca taccaacatg 19200
ccaaatgtga acgagttcat gtttaccaat aagtttaagg cgcgggtgat ggtgtcgcgc 19260
ttgcctacta aggacaatca ggtggagctg aaatacgagt gggtggagtt cacgctgccc 19320
gagggcaact actccgagac catgaccata gaccttatga acaacgcgat cgtggagcac 19380
tacttgaaag tgggcagaca gaacggggtt ctggaaagcg acatcggggt aaagtttgac 19440
acccgcaact tcagactggg gtttgacccc gtcactggtc ttgtcatgcc tggggtatat 19500
acaaacgaag ccttccatcc agacatcatt ttgctgccag gatgcggggt ggacttcacc 19560
cacagccgcc tgagcaactt gttgggcatc cgcaagcggc aacccttcca ggagggcttt 19620
aggatcacct acgatgatct ggagggtggt aacattcccg cactgttgga tgtggacgcc 19680
taccaggcga gcttgaaaga tgacaccgaa cagggcgggg gtggcgcagg cggcagcaac 19740
agcagtggca gcggcgcgga agagaactcc aacgcggcag ccgcggcaat gcagccggtg 19800
gaggacatga acgatcatgc cattcgcggc gacacctttg ccacacgggc tgaggagaag 19860
cgcgctgagg ccgaagcagc ggccgaagct gccgcccccg ctgcgcaacc cgaggtcgag 19920
aagcctcaga agaaaccggt gatcaaaccc ctgacagagg acagcaagaa acgcagttac 19980
aacctaataa gcaatgacag caccttcacc cagtaccgca gctggtacct tgcatacaac 20040
tacggcgacc ctcagaccgg aatccgctca tggaccctgc tttgcactcc tgacgtaacc 20100
tgcggctcgg agcaggtcta ctggtcgttg ccagacatga tgcaagaccc cgtgaccttc 20160
cgctccacgc gccagatcag caactttccg gtggtgggcg ccgagctgtt gcccgtgcac 20220
tccaagagct tctacaacga ccaggccgtc tactcccaac tcatccgcca gtttacctct 20280
ctgacccacg tgttcaatcg ctttcccgag aaccagattt tggcgcgccc gccagccccc 20340
accatcacca ccgtcagtga aaacgttcct gctctcacag atcacgggac gctaccgctg 20400
cgcaacagca tcggaggagt ccagcgagtg accattactg acgccagacg ccgcacctgc 20460
ccctacgttt acaaggccct gggcatagtc tcgccgcgcg tcctatcgag ccgcactttt 20520
tgagcaagca tgtccatcct tatatcgccc agcaataaca caggctgggg cctgcgcttc 20580
ccaagcaaga tgtttggcgg ggccaagaag cgctccgacc aacacccagt gcgcgtgcgc 20640
gggcactacc gcgcgccctg gggcgcgcac aaacgcggcc gcactgggcg caccaccgtc 20700
gatgacgcca tcgacgcggt ggtggaggag gcgcgcaact acacgcccac gccgccacca 20760
gtgtccacag tggacgcggc cattcagacc gtggtgcgcg gagcccggcg ctatgctaaa 20820
atgaagagac ggcggaggcg cgtagcacgt cgccaccgcc gccgacccgg cactgccgcc 20880
caacgcgcgg cggcggccct gcttaaccgc gcacgtcgca ccggccgacg ggcggccatg 20940
cgggccgctc gaaggctggc cgcgggtatt gtcactgtgc cccccaggtc caggcgacga 21000
gcggccgccg cagcagccgc ggccattagt gctatgactc agggtcgcag gggcaacgtg 21060
tattgggtgc gcgactcggt tagcggcctg cgcgtgcccg tgcgcacccg ccccccgcgc 21120
aactagattg caagaaaaaa ctacttagac tcgtactgtt gtatgtatcc agcggcggcg 21180
gcgcgcaacg aagctatgtc caagcgcaaa atcaaagaag agatgctcca ggtcatcgcg 21240
ccggagatct atggcccccc gaagaaggaa gagcaggatt acaagccccg aaagctaaag 21300
cgggtcaaaa agaaaaagaa agatgatgat gatgaacttg acgacgaggt ggaactgctg 21360
cacgctaccg cgcccaggcg acgggtacag tggaaaggtc gacgcgtaaa acgtgttttg 21420
cgacccggca ccaccgtagt ctttacgccc ggtgagcgct ccacccgcac ctacaagcgc 21480
gtgtatgatg aggtgtacgg cgacgaggac ctgcttgagc aggccaacga gcgcctcggg 21540
gagtttgcct acggaaagcg gcataaggac atgctggcgt tgccgctgga cgagggcaac 21600
ccaacaccta gcctaaagcc cgtaacactg cagcaggtgc tgcccgcgct tgcaccgtcc 21660
gaagaaaagc gcggcctaaa gcgcgagtct ggtgacttgg cacccaccgt gcagctgatg 21720
gtacccaagc gccagcgact ggaagatgtc ttggaaaaaa tgaccgtgga acctgggctg 21780
gagcccgagg tccgcgtgcg gccaatcaag caggtggcgc cgggactggg cgtgcagacc 21840
gtggacgttc agatacccac taccagtagc accagtattg ccaccgccac agagggcatg 21900
gagacacaaa cgtccccggt tgcctcagcg gtggcggatg ccgcggtgca ggcggtcgct 21960
gcggccgcgt ccaagacctc tacggaggtg caaacggacc cgtggatgtt tcgcgtttca 22020
gccccccggc gcccgcgcgg ttcgaggaag tacggcgccg ccagcgcgct actgcccgaa 22080
tatgccctac atccttccat tgcgcctacc cccggctatc gtggctacac ctaccgcccc 22140
agaagacgag caactacccg acgccgaacc accactggaa cccgccgccg ccgtcgccgt 22200
cgccagcccg tgctggcccc gatttccgtg cgcagggtgg ctcgcgaagg aggcaggacc 22260
ctggtgctgc caacagcgcg ctaccacccc agcatcgttt aaaagccggt ctttgtggtt 22320
cttgcagata tggccctcac ctgccgcctc cgtttcccgg tgccgggatt ccgaggaaga 22380
atgcaccgta ggaggggcat ggccggccac ggcctgacgg gcggcatgcg tcgtgcgcac 22440
caccggcggc ggcgcgcgtc gcaccgtcgc atgcgcggcg gtatcctgcc cctccttatt 22500
ccactgatcg ccgcggcgat tggcgccgtg cccggaattg catccgtggc cttgcaggcg 22560
cagagacact gattaaaaac aagttgcatg tggaaaaatc aaaataaaaa gtctggactc 22620
tcacgctcgc ttggtcctgt aactattttg tagaatggaa gacatcaact ttgcgtctct 22680
ggccccgcga cacggctcgc gcccgttcat gggaaactgg caagatatcg gcaccagcaa 22740
tatgagcggt ggcgccttca gctggggctc gctgtggagc ggcattaaaa atttcggttc 22800
caccgttaag aactatggca gcaaggcctg gaacagcagc acaggccaga tgctgaggga 22860
taagttgaaa gagcaaaatt tccaacaaaa ggtggtagat ggcctggcct ctggcattag 22920
cggggtggtg gacctggcca accaggcagt gcaaaataag attaacagta agcttgatcc 22980
ccgccctccc gtagaggagc ctccaccggc cgtggagaca gtgtctccag aggggcgtgg 23040
cgaaaagcgt ccgcgccccg acagggaaga aactctggtg acgcaaatag acgagcctcc 23100
ctcgtacgag gaggcactaa agcaaggcct gcccaccacc cgtcccatcg cgcccatggc 23160
taccggagtg ctgggccagc acacacccgt aacgctggac ctgcctcccc ccgccgacac 23220
ccagcagaaa cctgtgctgc caggcccgac cgccgttgtt gtaacccgtc ctagccgcgc 23280
gtccctgcgc cgcgccgcca gcggtccgcg atcgttgcgg cccgtagcca gtggcaactg 23340
gcaaagcaca ctgaacagca tcgtgggtct gggggtgcaa tccctgaagc gccgacgatg 23400
cttctgaata gctaacgtgt cgtatgtgtg tcatgtatgc gtccatgtcg ccgccagagg 23460
agctgctgag ccgccgcgcg cccgctttcc aagatggcta ccccttcgat gatgccgcag 23520
tggtcttaca tgcacatctc gggccaggac gcctcggagt acctgagccc cgggctggtg 23580
cagtttgccc gcgccaccga gacgtacttc agcctgaata acaagtttag aaaccccacg 23640
gtggcgccta cgcacgacgt gaccacagac cggtcccagc gtttgacgct gcggttcatc 23700
cctgtggacc gtgaggatac tgcgtactcg tacaaggcgc ggttcaccct agctgtgggt 23760
gataaccgtg tgctggacat ggcttccacg tactttgaca tccgcggcgt gctggacagg 23820
ggccctactt ttaagcccta ctctggcact gcctacaacg ccctggctcc caagggtgcc 23880
ccaaatcctt gcgaatggga tgaagctgct actgctcttg aaataaacct agaagaagag 23940
gacgatgaca acgaagacga agtagacgag caagctgagc agcaaaaaac tcacgtattt 24000
gggcaggcgc cttattctgg tataaatatt acaaaggagg gtattcaaat aggtgtcgaa 24060
ggtcaaacac ctaaatatgc cgataaaaca tttcaacctg aacctcaaat aggagaatct 24120
cagtggtacg aaactgaaat taatcatgca gctgggagag tccttaaaaa gactacccca 24180
atgaaaccat gttacggttc atatgcaaaa cccacaaatg aaaatggagg gcaaggcatt 24240
cttgtaaagc aacaaaatgg aaagctagaa agtcaagtgg aaatgcaatt tttctcaact 24300
actgaggcga ccgcaggcaa tggtgataac ttgactccta aagtggtatt gtacagtgaa 24360
gatgtagata tagaaacccc agacactcat atttcttaca tgcccactat taaggaaggt 24420
aactcacgag aactaatggg ccaacaatct atgcccaaca ggcctaatta cattgctttt 24480
agggacaatt ttattggtct aatgtattac aacagcacgg gtaatatggg tgttctggcg 24540
ggccaagcat cgcagttgaa tgctgttgta gatttgcaag acagaaacac agagctttca 24600
taccagcttt tgcttgattc cattggtgat agaaccaggt acttttctat gtggaatcag 24660
gctgttgaca gctatgatcc agatgttaga attattgaaa atcatggaac tgaagatgaa 24720
cttccaaatt actgctttcc actgggaggt gtgattaata cagagactct taccaaggta 24780
aaacctaaaa caggtcagga aaatggatgg gaaaaagatg ctacagaatt ttcagataaa 24840
aatgaaataa gagttggaaa taattttgcc atggaaatca atctaaatgc caacctgtgg 24900
agaaatttcc tgtactccaa catagcgctg tatttgcccg acaagctaaa gtacagtcct 24960
tccaacgtaa aaatttctga taacccaaac acctacgact acatgaacaa gcgagtggtg 25020
gctcccgggt tagtggactg ctacattaac cttggagcac gctggtccct tgactatatg 25080
gacaacgtca acccatttaa ccaccaccgc aatgctggcc tgcgctaccg ctcaatgttg 25140
ctgggcaatg gtcgctatgt gcccttccac atccaggtgc ctcagaagtt ctttgccatt 25200
aaaaacctcc ttctcctgcc gggctcatac acctacgagt ggaacttcag gaaggatgtt 25260
aacatggttc tgcagagctc cctaggaaat gacctaaggg ttgacggagc cagcattaag 25320
tttgatagca tttgccttta cgccaccttc ttccccatgg cccacaacac cgcctccacg 25380
cttgaggcca tgcttagaaa cgacaccaac gaccagtcct ttaacgacta tctctccgcc 25440
gccaacatgc tctaccctat acccgccaac gctaccaacg tgcccatatc catcccctcc 25500
cgcaactggg cggctttccg cggctgggcc ttcacgcgcc ttaagactaa ggaaacccca 25560
tcactgggct cgggctacga cccttattac acctactctg gctctatacc ctacctagat 25620
ggaacctttt acctcaacca cacctttaag aaggtggcca ttacctttga ctcttctgtc 25680
agctggcctg gcaatgaccg cctgcttacc cccaacgagt ttgaaattaa gcgctcagtt 25740
gacggggagg gttacaacgt tgcccagtgt aacatgacca aagactggtt cctggtacaa 25800
atgctagcta actacaacat tggctaccag ggcttctata tcccagagag ctacaaggac 25860
cgcatgtact ccttctttag aaacttccag cccatgagcc gtcaggtggt ggatgatact 25920
aaatacaagg actaccaaca ggtgggcatc ctacaccaac acaacaactc tggatttgtt 25980
ggctaccttg cccccaccat gcgcgaagga caggcctacc ctgctaactt cccctatccg 26040
cttataggca agaccgcagt tgacagcatt acccagaaaa agtttctttg cgatcgcacc 26100
ctttggcgca tcccattctc cagtaacttt atgtccatgg gcgcactcac agacctgggc 26160
caaaaccttc tctacgccaa ctccgcccac gcgctagaca tgacttttga ggtggatccc 26220
atggacgagc ccacccttct ttatgttttg tttgaagtct ttgacgtggt ccgtgtgcac 26280
cggccgcacc gcggcgtcat cgaaaccgtg tacctgcgca cgcccttctc ggccggcaac 26340
gccacaacat aaagaagcaa gcaacatcaa caacagctgc cgccatgggc tccagtgagc 26400
aggaactgaa agccattgtc aaagatcttg gttgtgggcc atattttttg ggcacctatg 26460
acaagcgctt tccaggcttt gtttctccac acaagctcgc ctgcgccata gtcaatacgg 26520
ccggtcgcga gactgggggc gtacactgga tggcctttgc ctggaacccg cactcaaaaa 26580
catgctacct ctttgagccc tttggctttt ctgaccagcg actcaagcag gtttaccagt 26640
ttgagtacga gtcactcctg cgccgtagcg ccattgcttc ttcccccgac cgctgtataa 26700
cgctggaaaa gtccacccaa agcgtacagg ggcccaactc ggccgcctgt ggactattct 26760
gctgcatgtt tctccacgcc tttgccaact ggccccaaac tcccatggat cacaacccca 26820
ccatgaacct tattaccggg gtacccaact ccatgctcaa cagtccccag gtacagccca 26880
ccctgcgtcg caaccaggaa cagctctaca gcttcctgga gcgccactcg ccctacttcc 26940
gcagccacag tgcgcagatt aggagcgcca cttctttttg tcacttgaaa aacatgtaaa 27000
aataatgtac tagagacact ttcaataaag gcaaatgctt ttatttgtac actctcgggt 27060
gattatttac ccccaccctt gccgtctgcg ccgtttaaaa atcaaagggg ttctgccgcg 27120
catcgctatg cgccactggc agggacacgt tgcgatactg gtgtttagtg ctccacttaa 27180
actcaggcac aaccatccgc ggcagctcgg tgaagttttc actccacagg ctgcgcacca 27240
tcaccaacgc gtttagcagg tcgggcgccg atatcttgaa gtcgcagttg gggcctccgc 27300
cctgcgcgcg cgagttgcga tacacagggt tgcagcactg gaacactatc agcgccgggt 27360
ggtgcacgct ggccagcacg ctcttgtcgg agatcagatc cgcgtccagg tcctccgcgt 27420
tgctcagggc gaacggagtc aactttggta gctgccttcc caaaaagggc gcgtgcccag 27480
gctttgagtt gcactcgcac cgtagtggca tcaaaaggtg accgtgcccg gtctgggcgt 27540
taggatacag cgcctgcata aaagccttga tctgcttaaa agccacctga gcctttgcgc 27600
cttcagagaa gaacatgccg caagacttgc cggaaaactg attggccgga caggccgcgt 27660
cgtgcacgca gcaccttgcg tcggtgttgg agatctgcac cacatttcgg ccccaccggt 27720
tcttcacgat cttggccttg ctagactgct ccttcagcgc gcgctgcccg ttttcgctcg 27780
tcacatccat ttcaatcacg tgctccttat ttatcataat gcttccgtgt agacacttaa 27840
gctcgccttc gatctcagcg cagcggtgca gccacaacgc gcagcccgtg ggctcgtgat 27900
gcttgtaggt cacctctgca aacgactgca ggtacgcctg caggaatcgc cccatcatcg 27960
tcacaaaggt cttgttgctg gtgaaggtca gctgcaaccc gcggtgctcc tcgttcagcc 28020
aggtcttgca tacggccgcc agagcttcca cttggtcagg cagtagtttg aagttcgcct 28080
ttagatcgtt atccacgtgg tacttgtcca tcagcgcgcg cgcagcctcc atgcccttct 28140
cccacgcaga cacgatcggc acactcagcg ggttcatcac cgtaatttca ctttccgctt 28200
cgctgggctc ttcctcttcc tcttgcgtcc gcataccacg cgccactggg tcgtcttcat 28260
tcagccgccg cactgtgcgc ttacctcctt tgccatgctt gattagcacc ggtgggttgc 28320
tgaaacccac catttgtagc gccacatctt ctctttcttc ctcgctgtcc acgattacct 28380
ctggtgatgg cgggcgctcg ggcttgggag aagggcgctt ctttttcttc ttgggcgcaa 28440
tggccaaatc cgccgccgag gtcgatggcc gcgggctggg tgtgcgcggc accagcgcgt 28500
cttgtgatga gtcttcctcg tcctcggact cgatacgccg cctcatccgc ttttttgggg 28560
gcgcccgggg aggcggcggc gacggggacg gggacgacac gtcctccatg gttgggggac 28620
gtcgcgccgc accgcgtccg cgctcggggg tggtttcgcg ctgctcctct tcccgactgg 28680
ccatttcctt ctcctatagg cagaaaaaga tcatggagtc agtcgagaag aaggacagcc 28740
taaccgcccc ctctgagttc gccaccaccg cctccaccga tgccgccaac gcgcctacca 28800
ccttccccgt cgaggcaccc ccgcttgagg aggaggaagt gattatcgag caggacccag 28860
gttttgtaag cgaagacgac gaggaccgct cagtaccaac agaggataaa aagcaagacc 28920
aggacaacgc agaggcaaac gaggaacaag tcgggcgggg ggacgaaagg catggcgact 28980
acctagatgt gggagacgac gtgctgttga agcatctgca gcgccagtgc gccattatct 29040
gcgacgcgtt gcaagagcgc agcgatgtgc ccctcgccat agcggatgtc agccttgcct 29100
acgaacgcca cctattctca ccgcgcgtac cccccaaacg ccaagaaaac ggcacatgcg 29160
agcccaaccc gcgcctcaac ttctaccccg tatttgccgt gccagaggtg cttgccacct 29220
atcacatctt tttccaaaac tgcaagatac ccctatcctg ccgtgccaac cgcagccgag 29280
cggacaagca gctggccttg cggcagggcg ctgtcatacc tgatatcgcc tcgctcaacg 29340
aagtgccaaa aatctttgag ggtcttggac gcgacgagaa gcgcgcggca aacgctctgc 29400
aacaggaaaa cagcgaaaat gaaagtcact ctggagtgtt ggtggaactc gagggtgaca 29460
acgcgcgcct agccgtacta aaacgcagca tcgaggtcac ccactttgcc tacccggcac 29520
ttaacctacc ccccaaggtc atgagcacag tcatgagtga gctgatcgtg cgccgtgcgc 29580
agcccctgga gagggatgca aatttgcaag aacaaacaga ggagggccta cccgcagttg 29640
gcgacgagca gctagcgcgc tggcttcaaa cgcgcgagcc tgccgacttg gaggagcgac 29700
gcaaactaat gatggccgca gtgctcgtta ccgtggagct tgagtgcatg cagcggttct 29760
ttgctgaccc ggagatgcag cgcaagctag aggaaacatt gcactacacc tttcgacagg 29820
gctacgtacg ccaggcctgc aagatctcca acgtggagct ctgcaacctg gtctcctacc 29880
ttggaatttt gcacgaaaac cgccttgggc aaaacgtgct tcattccacg ctcaagggcg 29940
aggcgcgccg cgactacgtc cgcgactgcg tttacttatt tctatgctac acctggcaga 30000
cggccatggg cgtttggcag cagtgcttgg aggagtgcaa cctcaaggag ctgcagaaac 30060
tgctaaagca aaacttgaag gacctatgga cggccttcaa cgagcgctcc gtggccgcgc 30120
acctggcgga catcattttc cccgaacgcc tgcttaaaac cctgcaacag ggtctgccag 30180
acttcaccag tcaaagcatg ttgcagaact ttaggaactt tatcctagag cgctcaggaa 30240
tcttgcccgc cacctgctgt gcacttccta gcgactttgt gcccattaag taccgcgaat 30300
gccctccgcc gctttggggc cactgctacc ttctgcagct agccaactac cttgcctacc 30360
actctgacat aatggaagac gtgagcggtg acggtctact ggagtgtcac tgtcgctgca 30420
acctatgcac cccgcaccgc tccctggttt gcaattcgca gctgcttaac gaaagtcaaa 30480
ttatcggtac ctttgagctg cagggtccct cgcctgacga aaagtccgcg gctccggggt 30540
tgaaactcac tccggggctg tggacgtcgg cttaccttcg caaatttgta cctgaggact 30600
accacgccca cgagattagg ttctacgaag accaatcccg cccgccaaat gcggagctta 30660
ccgcctgcgt cattacccag ggccacattc ttggccaatt gcaagccatc aacaaagccc 30720
gccaagagtt tctgctacga aagggacggg gggtttactt ggacccccag tccggcgagg 30780
agctcaaccc aatccccccg ccgccgcagc cctatcagca gcagccgcgg gcccttgctt 30840
cccaggatgg cacccaaaaa gaagctgcag ctgccgccgc cacccacgga cgaggaggaa 30900
tactgggaca gtcaggcaga ggaggttttg gacgaggagg aggaggacat gatggaagac 30960
tgggagagcc tagacgagga agcttccgag gtcgaagagg tgtcagacga aacaccgtca 31020
ccctcggtcg cattcccctc gccggcgccc cagaaatcgg caaccggttc cagcatggct 31080
acaacctccg ctcctcaggc gccgccggca ctgcccgttc gccgacccaa ccgtagatgg 31140
gacaccactg gaaccagggc cggtaagtcc aagcagccgc cgccgttagc ccaagagcaa 31200
caacagcgcc aaggctaccg ctcatggcgc gggcacaaga acgccatagt tgcttgcttg 31260
caagactgtg ggggcaacat ctccttcgcc cgccgctttc ttctctacca tcacggcgtg 31320
gccttccccc gtaacatcct gcattactac cgtcatctct acagcccata ctgcaccggc 31380
ggcagcggca gcggcagcaa cagcagcggc cacacagaag caaaggcgac cggatagcaa 31440
gactctgaca aagcccaaga aatccacagc ggcggcagca gcaggaggag gagcgctgcg 31500
tctggcgccc aacgaacccg tatcgacccg cgagcttaga aacaggattt ttcccactct 31560
gtatgctata tttcaacaga gcaggggcca agaacaagag ctgaaaataa aaaacaggtc 31620
tctgcgatcc ctcacccgca gctgcctgta tcacaaaagc gaagatcagc ttcggcgcac 31680
gctggaagac gcggaggctc tcttcagtaa atactgcgcg ctgactctta aggactagtt 31740
tcgcgccctt tctcaaattt aagcgcgaaa actacgtcat ctccagcggc cacacccggc 31800
gccagcacct gtcgtcagcg ccattatgag caaggaaatt cccacgccct acatgtggag 31860
ttaccagcca caaatgggac ttgcggctgg agctgcccaa gactactcaa cccgaataaa 31920
ctacatgagc gcgggacccc acatgatatc ccgggtcaac ggaatccgcg cccaccgaaa 31980
ccgaattctc ttggaacagg cggctattac caccacacct cgtaataacc ttaatccccg 32040
tagttggccc gctgccctgg tgtaccagga aagtcccgct cccaccactg tggtacttcc 32100
cagagacgcc caggccgaag ttcagatgac taactcaggg gcgcagcttg cgggcggctt 32160
tcgtcacagg gtgcggtcgc ccgggcaggg tataactcac ctgacaatca gagggcgagg 32220
tattcagctc aacgacgagt cggtgagctc ctcgcttggt ctccgtccgg acgggacatt 32280
tcagatcggc ggcgccggcc gtccttcatt cacgcctcgt caggcaatcc taactctgca 32340
gacctcgtcc tctgagccgc gctctggagg cattggaact ctgcaattta ttgaggagtt 32400
tgtgccatcg gtctacttta accccttctc gggacctccc ggccactatc cggatcaatt 32460
tattcctaac tttgacgcgg taaaggactc ggcggacggc tacgactgaa tgttaagtgg 32520
agaggcagag caactgcgcc tgaaacacct ggtccactgt cgccgccaca agtgctttgc 32580
ccgcgactcc ggtgagtttt gctactttga attgcccgag gatcatatcg agggcccggc 32640
gcacggcgtc cggcttaccg cccagggaga gcttgcccgt agcctgattc gggagtttac 32700
ccagcgcccc ctgctagttg agcgggacag gggaccctgt gttctcactg tgatttgcaa 32760
ctgtcctaac cttggattac atcaagatcc tctagttata actagagtac ccggggatct 32820
tattcccttt aactaataaa aaaaaataat aaagcatcac ttacttaaaa tcagttagca 32880
aatttctgtc cagtttattc agcagcacct ccttgccctc ctcccagctc tggtattgca 32940
gcttcctcct ggctgcaaac tttctccaca atctaaatgg aatgtcagtt tcctcctgtt 33000
cctgtccatc cgcacccact atcttcatgt tgttgcagat gaagcgcgca agaccgtctg 33060
aagatacctt caaccccgtg tatccatatg acacggaaac cggtcctcca actgtgcctt 33120
ttcttactcc tccctttgta tcccccaatg ggtttcaaga gagtccccct ggggtactct 33180
ctttgcgcct atccgaacct ctagttacct ccaatggcat gcttgcgctc aaaatgggca 33240
acggcctctc tctggacgag gccggcaacc ttacctccca aaatgtaacc actgtgagcc 33300
cacctctcaa aaaaaccaag tcaaacataa acctggaaat atctgcaccc ctcacagtta 33360
cctcagaagc cctaactgtg gctgccgccg cacctctaat ggtcgcgggc aacacactca 33420
ccatgcaatc acaggccccg ctaaccgtgc acgactccaa acttagcatt gccacccaag 33480
gacccctcac agtgtcagaa ggaaagctag ccctgcaaac atcaggcccc ctcaccacca 33540
ccgatagcag tacccttact atcactgcct caccccctct aactactgcc actggtagct 33600
tgggcattga cttgaaagag cccatttata cacaaaatgg aaaactagga ctaaagtacg 33660
gggctccttt gcatgtaaca gacgacctaa acactttgac cgtagcaact ggtccaggtg 33720
tgactattaa taatacttcc ttgcaaacta aagttactgg agccttgggt tttgattcac 33780
aaggcaatat gcaacttaat gtagcaggag gactaaggat tgattctcaa aacagacgcc 33840
ttatacttga tgttagttat ccgtttgatg ctcaaaacca actaaatcta agactaggac 33900
agggccctct ttttataaac tcagcccaca acttggatat taactacaac aaaggccttt 33960
acttgtttac agcttcaaac aattccaaaa agcttgaggt taacctaagc actgccaagg 34020
ggttgatgtt tgacgctaca gccatagcca ttaatgcagg agatgggctt gaatttggtt 34080
cacctaatgc accaaacaca aatcccctca aaacaaaaat tggccatggc ctagaatttg 34140
attcaaacaa ggctatggtt cctaaactag gaactggcct tagttttgac agcacaggtg 34200
ccattacagt aggaaacaaa aataatgata agctaacttt gtggaccaca ccagctccat 34260
ctcctaactg tagactaaat gcagagaaag atgctaaact cactttggtc ttaacaaaat 34320
gtggcagtca aatacttgct acagtttcag ttttggctgt taaaggcagt ttggctccaa 34380
tatctggaac agttcaaagt gctcatctta ttataagatt tgacgaaaat ggagtgctac 34440
taaacaattc cttcctggac ccagaatatt ggaactttag aaatggagat cttactgaag 34500
gcacagccta tacaaacgct gttggattta tgcctaacct atcagcttat ccaaaatctc 34560
acggtaaaac tgccaaaagt aacattgtca gtcaagttta cttaaacgga gacaaaacta 34620
aacctgtaac actaaccatt acactaaacg gtacacagga aacaggagac acaactccaa 34680
gtgcatactc tatgtcattt tcatgggact ggtctggcca caactacatt aatgaaatat 34740
ttgccacatc ctcttacact ttttcataca ttgcccaaga ataaagaatc gtttgtgtta 34800
tgtttcaacg tgtttatttt tcaattgcag aaaatttcaa gtcatttttc attcagtagt 34860
atagccccac caccacatag cttatacaga tcaccgtacc ttaatcaaac tcacagaacc 34920
ctagtattca acctgccacc tccctcccaa cacacagagt acacagtcct ttctccccgg 34980
ctggccttaa aaagcatcat atcatgggta acagacatat tcttaggtgt tatattccac 35040
acggtttcct gtcgagccaa acgctcatca gtgatattaa taaactcccc gggcagctca 35100
cttaagttca tgtcgctgtc cagctgctga gccacaggct gctgtccaac ttgcggttgc 35160
ttaacgggcg gcgaaggaga agtccacgcc tacatggggg tagagtcata atcgtgcatc 35220
aggatagggc ggtggtgctg cagcagcgcg cgaataaact gctgccgccg ccgctccgtc 35280
ctgcaggaat acaacatggc agtggtctcc tcagcgatga ttcgcaccgc ccgcagcata 35340
aggcgccttg tcctccgggc acagcagcgc accctgatct cacttaaatc agcacagtaa 35400
ctgcagcaca gcaccacaat attgttcaaa atcccacagt gcaaggcgct gtatccaaag 35460
ctcatggcgg ggaccacaga acccacgtgg ccatcatacc acaagcgcag gtagattaag 35520
tggcgacccc tcataaacac gctggacata aacattacct cttttggcat gttgtaattc 35580
accacctccc ggtaccatat aaacctctga ttaaacatgg cgccatccac caccatccta 35640
aaccagctgg ccaaaacctg cccgccggct atacactgca gggaaccggg actggaacaa 35700
tgacagtgga gagcccagga ctcgtaacca tggatcatca tgctcgtcat gatatcaatg 35760
ttggcacaac acaggcacac gtgcatacac ttcctcagga ttacaagctc ctcccgcgtt 35820
agaaccatat cccagggaac aacccattcc tgaatcagcg taaatcccac actgcaggga 35880
agacctcgca cgtaactcac gttgtgcatt gtcaaagtgt tacattcggg cagcagcgga 35940
tgatcctcca gtatggtagc gcgggtttct gtctcaaaag gaggtagacg atccctactg 36000
tacggagtgc gccgagacaa ccgagatcgt gttggtcgta gtgtcatgcc aaatggaacg 36060
ccggacgtag tcatatttcc tgaagcaaaa ccaggtgcgg gcgtgacaaa cagatctgcg 36120
tctccggtct cgccgcttag atcgctctgt gtagtagttg tagtatatcc actctctcaa 36180
agcatccagg cgccccctgg cttcgggttc tatgtaaact ccttcatgcg ccgctgccct 36240
gataacatcc accaccgcag aataagccac acccagccaa cctacacatt cgttctgcga 36300
gtcacacacg ggaggagcgg gaagagctgg aagaaccatg tttttttttt tattccaaaa 36360
gattatccaa aacctcaaaa tgaagatcta ttaagtgaac gcgctcccct ccggtggcgt 36420
ggtcaaactc tacagccaaa gaacagataa tggcatttgt aagatgttgc acaatggctt 36480
ccaaaaggca aacggccctc acgtccaagt ggacgtaaag gctaaaccct tcagggtgaa 36540
tctcctctat aaacattcca gcaccttcaa ccatgcccaa ataattctca tctcgccacc 36600
ttctcaatat atctctaagc aaatcccgaa tattaagtcc ggccattgta aaaatctgct 36660
ccagagcgcc ctccaccttc agcctcaagc agcgaatcat gattgcaaaa attcaggttc 36720
ctcacagacc tgtataagat tcaaaagcgg aacattaaca aaaataccgc gatcccgtag 36780
gtcccttcgc agggccagct gaacataatc gtgcaggtct gcacggacca gcgcggccac 36840
ttccccgcca ggaaccttga caaaagaacc cacactgatt atgacacgca tactcggagc 36900
tatgctaacc agcgtagccc cgatgtaagc tttgttgcat gggcggcgat ataaaatgca 36960
aggtgctgct caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca tcgtagtcat 37020
gctcatgcag ataaaggcag gtaagctccg gaaccaccac agaaaaagac accatttttc 37080
tctcaaacat gtctgcgggt ttctgcataa acacaaaata aaataacaaa aaaacattta 37140
aacattagaa gcctgtctta caacaggaaa aacaaccctt ataagcataa gacggactac 37200
ggccatgccg gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca ccaccgacag 37260
ctcctcggtc atgtccggag tcataatgta agactcggta aacacatcag gttgattcat 37320
cggtcagtgc taaaaagcga ccgaaatagc ccgggggaat acatacccgc aggcgtagag 37380
acaacattac agcccccata ggaggtataa caaaattaat aggagagaaa aacacataaa 37440
cacctgaaaa accctcctgc ctaggcaaaa tagcaccctc ccgctccaga acaacataca 37500
gcgcttcaca gcggcagcct aacagtcagc cttaccagta aaaaagaaaa cctattaaaa 37560
aaacaccact cgacacggca ccagctcaat cagtcacagt gtaaaaaagg gccaagtgca 37620
gagcgagtat atataggact aaaaaatgac gtaacggtta aagtccacaa aaaacaccca 37680
gaaaaccgca cgcgaaccta cgcccagaaa cgaaagccaa aaaacccaca acttcctcaa 37740
atcgtcactt ccgttttccc acgttacgta acttcccatt ttaagaaaac tacaattccc 37800
aacacataca agttactccg ccctaaaacc tacgtcaccc gccccgttcc cacgccccgc 37860
gccacgtcac aaactccacc ccctcattat catattggct tcaatccaaa ataaggtata 37920
ttattgatga tact 37934
<211> 37892
<212> DNA
<213> Artificial(人工序列)
ctttaattaa ggatcccgac ctgtcctcga ccgatgccct tgagagcctt caacccagtc 60
agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt 120
atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc 180
tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc 240
ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt 300
atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc 360
tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg 420
caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc 480
gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc 540
gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc 600
tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc 660
ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga 720
gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca 780
gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg 840
tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg 900
aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag 960
caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag 1020
cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa 1080
cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc 1140
gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat 1200
catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga 1260
acagaaattc ccccttacac ggaggcatca agtgaccaaa caggaaaaaa ccgcccttaa 1320
catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga 1380
cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg 1440
cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga 1500
gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc 1560
agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt 1620
gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 1680
tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 1740
tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 1800
aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 1860
aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 1920
ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 1980
acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2040
ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2100
tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 2160
tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 2220
gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 2280
agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 2340
tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 2400
agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 2460
tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 2520
acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 2580
tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 2640
agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 2700
tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 2760
acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 2820
tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 2880
ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 2940
agtagttcgc cagttaatag tttgcgcaac gttgttgtca aaaaggatct tcacctagat 3000
ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 3060
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 3120
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 3180
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctc 3240
gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag gatgaggatc 3300
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 3360
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 3420
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 3480
tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 3540
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 3600
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 3660
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 3720
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 3780
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat 3840
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 3900
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 3960
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 4020
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 4080
ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc tcatttttta 4140
accaataggc cgaaatcggc aacatccctt ataaatcaaa agaatagacc gcgatagggt 4200
tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 4260
aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa 4320
gttttttgcg gtcgaggtgc cgtaaagctc taaatcggaa ccctaaaggg agcccccgat 4380
ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 4440
gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 4500
cgcgcttaat gcgccgccgg attaattaat cagtcccttc cagctctctg ccccttttgg 4560
attgaagcca atatgataat gagggggtgg agtttgtgac gtggcgcggg gcgtgggaac 4620
ggggcgggtg acgtagtagt gtggcggaag tgtgatgttg caagtgtggc ggaacacatg 4680
taagcgacgg atgtggcaaa agtgacgttt ttggtgtgcg ccggtgtaca caggaagtga 4740
caattttcgc gcggttttag gcggatgttg tagtaaattt gggcgtaacc gagtaagatt 4800
tggccatttt cgcgggaaaa ctgaataaga ggaagtgaaa tctgaataat tttgtgttac 4860
tcatagcgcg taatcggtag ttattaatag taatcaatta cggggtcatt agttcatagc 4920
ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 4980
aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 5040
actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 5100
caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 5160
tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 5220
ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 5280
cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 5340
tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 5400
atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt 5460
cagatccgct agaccatggc ctcctccgag gacgtcatca aggagttcat gcgcttcaag 5520
gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc 5580
cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc 5640
ttcgcctggg acatcctgtc ccctcagttc cagtacggct ccaaggccta cgtgaagcac 5700
cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc 5760
gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac 5820
ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta 5880
atgcagaaga agaccatggg ctgggaggcc tccaccgagc ggatgtaccc cgaggacggc 5940
gccctgaagg gcgagatcaa gatgaggctg aagctgaagg acggcggcca ctacgacgcc 6000
gaggtcaaga ccacctacat ggccaagaag cccgtgcagc tgcccggcgc ctacaagacc 6060
gacatcaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag 6120
cgcgccgagg gccgccactc caccggcgcc aatccaccgg atctagataa ctgatcataa 6180
tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 6240
tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 6300
atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 6360
attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtctct agttattaat 6420
agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 6480
ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 6540
tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 6600
atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 6660
ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 6720
gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 6780
ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 6840
tccaccccat tgacgtcaat gggagtttgt tttggaacca aaatcaacgg gactttccaa 6900
aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 6960
tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc 7020
tccctatcag tgatagagat ctccctatca gtgatagaga tcgagctgtt tagtgaaccg 7080
tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgaattca 7140
ggggatccaa tcggaaagcg gacgcggaat ttaaattgat ttttgcggta taagaatata 7200
tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca 7260
gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg 7320
gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag 7380
cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg 7440
ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc 7500
cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg 7560
gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg 7620
gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg 7680
gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac 7740
gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct 7800
gcagctcgct cttcatttaa atcgagtatc ccgtgagcgc tttctagaga tatcggccgc 7860
taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 7920
tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 7980
gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 8040
caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 8100
tctatggctt ctgaggcgga aagaaccagg atctgctagg atctcgtaag ggtgggaaag 8160
aatatataag gtgggggtct tatgtagttt tgtatctgtt ttgcagcagc cgccgccgcc 8220
atgagcacca actcgtttga tggaagcatt gtgagctcat atttgacaac gcgcatgccc 8280
ccatgggccg gggtgcgtca gaatgtgatg ggctccagca ttgatggtcg ccccgtcctg 8340
cccgcaaact ctactacctt gacctacgag accgtgtctg gaacgccgtt ggagactgca 8400
gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac tgactttgct 8460
ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga tgacaagttg 8520
acggctcttt tggcacaatt ggattctttg acccgggaac ttaatgtcgt ttctcagcag 8580
ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt 8640
taaaacataa ataaaaaacc agactctgtt tggatttgga tcaagcaagt gtcttgctgt 8700
ctttatttag gggttttgcg cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg 8760
gtcctgtgta ttttttccag gacgtggtaa aggtgactct ggatgttcag atacatgggc 8820
ataagcccgt ctctggggtg gaggtagcac cactgcagag cttcatgctg cggggtggtg 8880
ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt 8940
agcaagctga ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg 9000
gatgggtgca tacgtgggga tatgagatgc atcttggact gtatttttag gttggctatg 9060
ttcccagcca tatccctccg gggattcatg ttgtgcagaa ccaccagcac agtgtatccg 9120
gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa cttggagacg 9180
cccttgtgac ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg 9240
gcggcggcct gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg 9300
agatcgtcat aggccatttt tacaaagcgc gggcggaggg tgccagactg cggtataatg 9360
gttccatccg gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt 9420
tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtaggg 9480
gagatcagct gggaagaaag caggttcctg agcagctgcg acttaccgca gccggtgggc 9540
ccgtaaatca cacctattac cgggtgcaac tggtagttaa gagagctgca gctgccgtca 9600
tccctgagca ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc 9660
aaatccgcca gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt 9720
ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc 9780
aggcggtccc acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt 9840
ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca 9900
gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg 9960
ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga 10020
agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt 10080
ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcacg 10140
aggggcagtg cagacttttg agggcgtaga gcttgggcgc gagaaatacc gattccgggg 10200
agtaggcatc cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct 10260
ctggccgttc ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc 10320
tggtttccat gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata 10380
cagacttgag aggcctgtcc tcgagcggtg ttccgcggtc ctcctcgtat agaaactcgg 10440
accactctga gacaaaggct cgcgtccagg ccagcacgaa ggaggctaag tgggaggggt 10500
agcggtcgtt gtccactagg gggtccactc gctccagggt gtgaagacac atgtcgccct 10560
cttcggcatc aaggaaggtg attggtttgt aggtgtaggc cacgtgaccg ggtgttcctg 10620
aaggggggct ataaaagggg gtgggggcgc gttcgtcctc actctcttcc gcatcgctgt 10680
ctgcgagggc cagctgttgg ggtgagtact ccctctgaaa agcgggcatg acttctgcgc 10740
taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc gcggtgatgc 10800
ctttgagggt ggccgcatcc atctggtcag aaaagacaat ctttttgttg tcaagcttgg 10860
tggcaaacga cccgtagagg gcgttggaca gcaacttggc gatggagcgc agggtttggt 10920
ttttgtcgcg atcggcgcgc tccttggccg cgatgtttag ctgcacgtat tcgcgcgcaa 10980
cgcaccgcca ttcgggaaag acggtggtgc gctcgtcggg caccaggtgc acgcgccaac 11040
cgcggttgtg cagggtgaca aggtcaacgc tggtggctac ctctccgcgt aggcgctcgt 11100
tggtccagca gaggcggccg cccttgcgcg agcagaatgg cggtaggggg tctagctgcg 11160
tctcgtccgg ggggtctgcg tccacggtaa agaccccggg cagcaggcgc gcgtcgaagt 11220
agtctatctt gcatccttgc aagtctagcg cctgctgcca tgcgcgggcg gcaagcgcgc 11280
gctcgtatgg gttgagtggg ggaccccatg gcatggggtg ggtgagcgcg gaggcgtaca 11340
tgccgcaaat gtcgtaaacg tagaggggct ctctgagtat tccaagatat gtagggtagc 11400
atcttccacc gcggatgctg gcgcgcacgt aatcgtatag ttcgtgcgag ggagcgagga 11460
ggtcgggacc gaggttgcta cgggcgggct gctctgctcg gaagactatc tgcctgaaga 11520
tggcatgtga gttggatgat atggttggac gctggaagac gttgaagctg gcgtctgtga 11580
gacctaccgc gtcacgcacg aaggaggcgt aggagtcgcg cagcttgttg accagctcgg 11640
cggtgacctg cacgtctagg gcgcagtagt ccagggtttc cttgatgatg tcatacttat 11700
cctgtccctt ttttttccac agctcgcggt tgaggacaaa ctcttcgcgg tctttccagt 11760
actcttggat cggaaacccg tcggcctccg aacggtaaga gcctagcatg tagaactggt 11820
tgacggcctg gtaggcgcag catccctttt ctacgggtag cgcgtatgcc tgcgcggcct 11880
tccggagcga ggtgtgggtg agcgcaaagg tgtccctgac catgactttg aggtactggt 11940
atttgaagtc agtgtcgtcg catccgccct gctcccagag caaaaagtcc gtgcgctttt 12000
tggaacgcgg atttggcagg gcgaaggtga catcgttgaa gagtatcttt cccgcgcgag 12060
gcataaagtt gcgtgtgatg cggaagggtc ccggcacctc ggaacggttg ttaattacct 12120
gggcggcgag cacgatctcg tcaaagccgt tgatgttgtg gcccacaatg taaagttcca 12180
agaagcgcgg gatgcccttg atggaaggca attttttaag ttcctcgtag gtgagctctt 12240
caggggagct gagcccgtgc tctgaaaggg cccagtctgc aagatgaggg ttggaagcga 12300
cgaatgagct ccacaggtca cgggccatta gcatttgcag gtggtcgcga aaggtcctaa 12360
actggcgacc tatggccatt ttttctgggg tgatgcagta gaaggtaagc gggtcttgtt 12420
cccagcggtc ccatccaagg ttcgcggcta ggtctcgcgc ggcagtcact agaggctcat 12480
ctccgccgaa cttcatgacc agcatgaagg gcacgagctg cttcccaaag gcccccatcc 12540
aagtataggt ctctacatcg taggtgacaa agagacgctc ggtgcgagga tgcgagccga 12600
tcgggaagaa ctggatctcc cgccaccaat tggaggagtg gctattgatg tggtgaaagt 12660
agaagtccct gcgacgggcc gaacactcgt gctggctttt gtaaaaacgt gcgcagtact 12720
ggcagcggtg cacgggctgt acatcctgca cgaggttgac ctgacgaccg cgcacaagga 12780
agcagagtgg gaatttgagc ccctcgcctg gcgggtttgg ctggtggtct tctacttcgg 12840
ctgcttgtcc ttgaccgtct ggctgctcga ggggagttac ggtggatcgg accaccacgc 12900
cgcgcgagcc caaagtccag atgtccgcgc gcggcggtcg gagcttgatg acaacatcgc 12960
gcagatggga gctgtccatg gtctggagct cccgcggcgt caggtcaggc gggagctcct 13020
gcaggtttac ctcgcataga cgggtcaggg cgcgggctag atccaggtga tacctaattt 13080
ccaggggctg gttggtggcg gcgtcgatgg cttgcaagag gccgcatccc cgcggcgcga 13140
ctacggtacc gcgcggcggg cggtgggccg cgggggtgtc cttggatgat gcatctaaaa 13200
gcggtgacgc gggcgagccc ccggaggtag ggggggctcc ggacccgccg ggagaggggg 13260
caggggcacg tcggcgccgc gcgcgggcag gagctggtgc tgcgcgcgta ggttgctggc 13320
gaacgcgacg acgcggcggt tgatctcctg aatctggcgc ctctgcgtga agacgacggg 13380
cccggtgagc ttgagcctga aagagagttc gacagaatca atttcggtgt cgttgacggc 13440
ggcctggcgc aaaatctcct gcacgtctcc tgagttgtct tgataggcga tctcggccat 13500
gaactgctcg atctcttcct cctggagatc tccgcgtccg gctcgctcca cggtggcggc 13560
gaggtcgttg gaaatgcggg ccatgagctg cgagaaggcg ttgaggcctc cctcgttcca 13620
gacgcggctg tagaccacgc ccccttcggc atcgcgggcg cgcatgacca cctgcgcgag 13680
attgagctcc acgtgccggg cgaagacggc gtagtttcgc aggcgctgaa agaggtagtt 13740
gagggtggtg gcggtgtgtt ctgccacgaa gaagtacata acccagcgtc gcaacgtgga 13800
ttcgttgata tcccccaagg cctcaaggcg ctccatggcc tcgtagaagt ccacggcgaa 13860
gttgaaaaac tgggagttgc gcgccgacac ggttaactcc tcctccagaa gacggatgag 13920
ctcggcgaca gtgtcgcgca cctcgcgctc aaaggctaca ggggcctctt cttcttcttc 13980
aatctcctct tccataaggg cctccccttc ttcttcttct ggcggcggtg ggggaggggg 14040
gacacggcgg cgacgacggc gcaccgggag gcggtcgaca aagcgctcga tcatctcccc 14100
gcggcgacgg cgcatggtct cggtgacggc gcggccgttc tcgcgggggc gcagttggaa 14160
gacgccgccc gtcatgtccc ggttatgggt tggcgggggg ctgccatgcg gcagggatac 14220
ggcgctaacg atgcatctca acaattgttg tgtaggtact ccgccgccga gggacctgag 14280
cgagtccgca tcgaccggat cggaaaacct ctcgagaaag gcgtctaacc agtcacagtc 14340
gcaaggtagg ctgagcaccg tggcgggcgg cagcgggcgg cggtcggggt tgtttctggc 14400
ggaggtgctg ctgatgatgt aattaaagta ggcggtcttg agacggcgga tggtcgacag 14460
aagcaccatg tccttgggtc cggcctgctg aatgcgcagg cggtcggcca tgccccaggc 14520
ttcgttttga catcggcgca ggtctttgta gtagtcttgc atgagccttt ctaccggcac 14580
ttcttcttct ccttcctctt gtcctgcatc tcttgcatct atcgctgcgg cggcggcgga 14640
gtttggccgt aggtggcgcc ctcttcctcc catgcgtgtg accccgaagc ccctcatcgg 14700
ctgaagcagg gctaggtcgg cgacaacgcg ctcggctaat atggcctgct gcacctgcgt 14760
gagggtagac tggaagtcat ccatgtccac aaagcggtgg tatgcgcccg tgttgatggt 14820
gtaagtgcag ttggccataa cggaccagtt aacggtctgg tgacccggct gcgagagctc 14880
ggtgtacctg agacgcgagt aagccctcga gtcaaatacg tagtcgttgc aagtccgcac 14940
caggtactgg tatcccacca aaaagtgcgg cggcggctgg cggtagaggg gccagcgtag 15000
ggtggccggg gctccggggg cgagatcttc caacataagg cgatgatatc cgtagatgta 15060
cctggacatc caggtgatgc cggcggcggt ggtggaggcg cgcggaaagt cgcggacgcg 15120
gttccagatg ttgcgcagcg gcaaaaagtg ctccatggtc gggacgctct ggccggtcag 15180
gcgcgcgcaa tcgttgacgc tctaccgtgc aaaaggagag cctgtaagcg ggcactcttc 15240
cgtggtctgg tggataaatt cgcaagggta tcatggcgga cgaccggggt tcgagccccg 15300
tatccggccg tccgccgtga tccatgcggt taccgcccgc gtgtcgaacc caggtgtgcg 15360
acgtcagaca acgggggagt gctccttttg gcttccttcc aggcgcggcg gctgctgcgc 15420
tagctttttt ggccactggc cgcgcgcagc gtaagcggtt aggctggaaa gcgaaagcat 15480
taagtggctc gctccctgta gccggagggt tattttccaa gggttgagtc gcgggacccc 15540
cggttcgagt ctcggaccgg ccggactgcg gcgaacgggg gtttgcctcc ccgtcatgca 15600
agaccccgct tgcaaattcc tccggaaaca gggacgagcc ccttttttgc ttttcccaga 15660
tgcatccggt gctgcggcag atgcgccccc ctcctcagca gcggcaagag caagagcagc 15720
ggcagacatg cagggcaccc tcccctcctc ctaccgcgtc aggaggggcg acatccgcgg 15780
ttgacgcggc agcagatggt gattacgaac ccccgcggcg ccgggcccgg cactacctgg 15840
acttggagga gggcgagggc ctggcgcggc taggagcgcc ctctcctgag cggtacccaa 15900
gggtgcagct gaagcgtgat acgcgtgagg cgtacgtgcc gcggcagaac ctgtttcgcg 15960
accgcgaggg agaggagccc gaggagatgc gggatcgaaa gttccacgca gggcgcgagc 16020
tgcggcatgg cctgaatcgc gagcggttgc tgcgcgagga ggactttgag cccgacgcgc 16080
gaaccgggat tagtcccgcg cgcgcacacg tggcggccgc cgacctggta accgcatacg 16140
agcagacggt gaaccaggag attaactttc aaaaaagctt taacaaccac gtgcgtacgc 16200
ttgtggcgcg cgaggaggtg gctataggac tgatgcatct gtgggacttt gtaagcgcgc 16260
tggagcaaaa cccaaatagc aagccgctca tggcgcagct gttccttata gtgcagcaca 16320
gcagggacaa cgaggcattc agggatgcgc tgctaaacat agtagagccc gagggccgct 16380
ggctgctcga tttgataaac atcctgcaga gcatagtggt gcaggagcgc agcttgagcc 16440
tggctgacaa ggtggccgcc atcaactatt ccatgcttag cctgggcaag ttttacgccc 16500
gcaagatata ccatacccct tacgttccca tagacaagga ggtaaagatc gaggggttct 16560
acatgcgcat ggcgctgaag gtgcttacct tgagcgacga cctgggcgtt tatcgcaacg 16620
agcgcatcca caaggccgtg agcgtgagcc ggcggcgcga gctcagcgac cgcgagctga 16680
tgcacagcct gcaaagggcc ctggctggca cgggcagcgg cgatagagag gccgagtcct 16740
actttgacgc gggcgctgac ctgcgctggg ccccaagccg acgcgccctg gaggcagctg 16800
gggccggacc tgggctggcg gtggcacccg cgcgcgctgg caacgtcggc ggcgtggagg 16860
aatatgacga ggacgatgag tacgagccag aggacggcga gtactaagcg gtgatgtttc 16920
tgatcagatg atgcaagacg caacggaccc ggcggtgcgg gcggcgctgc agagccagcc 16980
gtccggcctt aactccacgg acgactggcg ccaggtcatg gaccgcatca tgtcgctgac 17040
tgcgcgcaat cctgacgcgt tccggcagca gccgcaggcc aaccggctct ccgcaattct 17100
ggaagcggtg gtcccggcgc gcgcaaaccc cacgcacgag aaggtgctgg cgatcgtaaa 17160
cgcgctggcc gaaaacaggg ccatccggcc cgacgaggcc ggcctggtct acgacgcgct 17220
gcttcagcgc gtggctcgtt acaacagcgg caacgtgcag accaacctgg accggctggt 17280
gggggatgtg cgcgaggccg tggcgcagcg tgagcgcgcg cagcagcagg gcaacctggg 17340
ctccatggtt gcactaaacg ccttcctgag tacacagccc gccaacgtgc cgcggggaca 17400
ggaggactac accaactttg tgagcgcact gcggctaatg gtgactgaga caccgcaaag 17460
tgaggtgtac cagtctgggc cagactattt tttccagacc agtagacaag gcctgcagac 17520
cgtaaacctg agccaggctt tcaaaaactt gcaggggctg tggggggtgc gggctcccac 17580
aggcgaccgc gcgaccgtgt ctagcttgct gacgcccaac tcgcgcctgt tgctgctgct 17640
aatagcgccc ttcacggaca gtggcagcgt gtcccgggac acatacctag gtcacttgct 17700
gacactgtac cgcgaggcca taggtcaggc gcatgtggac gagcatactt tccaggagat 17760
tacaagtgtc agccgcgcgc tggggcagga ggacacgggc agcctggagg caaccctaaa 17820
ctacctgctg accaaccggc ggcagaagat cccctcgttg cacagtttaa acagcgagga 17880
ggagcgcatt ttgcgctacg tgcagcagag cgtgagcctt aacctgatgc gcgacggggt 17940
aacgcccagc gtggcgctgg acatgaccgc gcgcaacatg gaaccgggca tgtatgcctc 18000
aaaccggccg tttatcaacc gcctaatgga ctacttgcat cgcgcggccg ccgtgaaccc 18060
cgagtatttc accaatgcca tcttgaaccc gcactggcta ccgccccctg gtttctacac 18120
cgggggattc gaggtgcccg agggtaacga tggattcctc tgggacgaca tagacgacag 18180
cgtgttttcc ccgcaaccgc agaccctgct agagttgcaa cagcgcgagc aggcagaggc 18240
ggcgctgcga aaggaaagct tccgcaggcc aagcagcttg tccgatctag gcgctgcggc 18300
cccgcggtca gatgctagta gcccatttcc aagcttgata gggtctctta ccagcactcg 18360
caccacccgc ccgcgcctgc tgggcgagga ggagtaccta aacaactcgc tgctgcagcc 18420
gcagcgcgaa aaaaacctgc ctccggcatt tcccaacaac gggatagaga gcctagtgga 18480
caagatgagt agatggaaga cgtacgcgca ggagcacagg gacgtgccag gcccgcgccc 18540
gcccacccgt cgtcaaaggc acgaccgtca gcggggtctg gtgtgggagg acgatgactc 18600
ggcagacgac agcagcgtcc tggatttggg agggagtggc aacccgtttg cgcaccttcg 18660
ccccaggctg gggagaatgt tttaaaaaaa aaaaagcatg atgcaaaata aaaaactcac 18720
caaggccatg gcaccgagcg ttggttttct tgtattcccc ttagtatgcg gcgcgcggcg 18780
atgtatgagg aaggtcctcc tccctcctac gagagtgtgg tgagcgcggc gccagtggcg 18840
gcggcgctgg gttctccctt cgatgctccc ctggacccgc cgtttgtgcc tccgcggtac 18900
ctgcggccta ccggggggag aaacagcatc cgttactctg agttggcacc cctattcgac 18960
accacccgtg tgtacctggt ggacaacaag tcaacggatg tggcatccct gaactaccag 19020
aacgaccaca gcaactttct gaccacggtc attcaaaaca atgactacag cccgggggag 19080
gcaagcacac agaccatcaa tcttgacgac cggtcgcact ggggcggcga cctgaaaacc 19140
atcctgcata ccaacatgcc aaatgtgaac gagttcatgt ttaccaataa gtttaaggcg 19200
cgggtgatgg tgtcgcgctt gcctactaag gacaatcagg tggagctgaa atacgagtgg 19260
gtggagttca cgctgcccga gggcaactac tccgagacca tgaccataga ccttatgaac 19320
aacgcgatcg tggagcacta cttgaaagtg ggcagacaga acggggttct ggaaagcgac 19380
atcggggtaa agtttgacac ccgcaacttc agactggggt ttgaccccgt cactggtctt 19440
gtcatgcctg gggtatatac aaacgaagcc ttccatccag acatcatttt gctgccagga 19500
tgcggggtgg acttcaccca cagccgcctg agcaacttgt tgggcatccg caagcggcaa 19560
cccttccagg agggctttag gatcacctac gatgatctgg agggtggtaa cattcccgca 19620
ctgttggatg tggacgccta ccaggcgagc ttgaaagatg acaccgaaca gggcgggggt 19680
ggcgcaggcg gcagcaacag cagtggcagc ggcgcggaag agaactccaa cgcggcagcc 19740
gcggcaatgc agccggtgga ggacatgaac gatcatgcca ttcgcggcga cacctttgcc 19800
acacgggctg aggagaagcg cgctgaggcc gaagcagcgg ccgaagctgc cgcccccgct 19860
gcgcaacccg aggtcgagaa gcctcagaag aaaccggtga tcaaacccct gacagaggac 19920
agcaagaaac gcagttacaa cctaataagc aatgacagca ccttcaccca gtaccgcagc 19980
tggtaccttg catacaacta cggcgaccct cagaccggaa tccgctcatg gaccctgctt 20040
tgcactcctg acgtaacctg cggctcggag caggtctact ggtcgttgcc agacatgatg 20100
caagaccccg tgaccttccg ctccacgcgc cagatcagca actttccggt ggtgggcgcc 20160
gagctgttgc ccgtgcactc caagagcttc tacaacgacc aggccgtcta ctcccaactc 20220
atccgccagt ttacctctct gacccacgtg ttcaatcgct ttcccgagaa ccagattttg 20280
gcgcgcccgc cagcccccac catcaccacc gtcagtgaaa acgttcctgc tctcacagat 20340
cacgggacgc taccgctgcg caacagcatc ggaggagtcc agcgagtgac cattactgac 20400
gccagacgcc gcacctgccc ctacgtttac aaggccctgg gcatagtctc gccgcgcgtc 20460
ctatcgagcc gcactttttg agcaagcatg tccatcctta tatcgcccag caataacaca 20520
ggctggggcc tgcgcttccc aagcaagatg tttggcgggg ccaagaagcg ctccgaccaa 20580
cacccagtgc gcgtgcgcgg gcactaccgc gcgccctggg gcgcgcacaa acgcggccgc 20640
actgggcgca ccaccgtcga tgacgccatc gacgcggtgg tggaggaggc gcgcaactac 20700
acgcccacgc cgccaccagt gtccacagtg gacgcggcca ttcagaccgt ggtgcgcgga 20760
gcccggcgct atgctaaaat gaagagacgg cggaggcgcg tagcacgtcg ccaccgccgc 20820
cgacccggca ctgccgccca acgcgcggcg gcggccctgc ttaaccgcgc acgtcgcacc 20880
ggccgacggg cggccatgcg ggccgctcga aggctggccg cgggtattgt cactgtgccc 20940
cccaggtcca ggcgacgagc ggccgccgca gcagccgcgg ccattagtgc tatgactcag 21000
ggtcgcaggg gcaacgtgta ttgggtgcgc gactcggtta gcggcctgcg cgtgcccgtg 21060
cgcacccgcc ccccgcgcaa ctagattgca agaaaaaact acttagactc gtactgttgt 21120
atgtatccag cggcggcggc gcgcaacgaa gctatgtcca agcgcaaaat caaagaagag 21180
atgctccagg tcatcgcgcc ggagatctat ggccccccga agaaggaaga gcaggattac 21240
aagccccgaa agctaaagcg ggtcaaaaag aaaaagaaag atgatgatga tgaacttgac 21300
gacgaggtgg aactgctgca cgctaccgcg cccaggcgac gggtacagtg gaaaggtcga 21360
cgcgtaaaac gtgttttgcg acccggcacc accgtagtct ttacgcccgg tgagcgctcc 21420
acccgcacct acaagcgcgt gtatgatgag gtgtacggcg acgaggacct gcttgagcag 21480
gccaacgagc gcctcgggga gtttgcctac ggaaagcggc ataaggacat gctggcgttg 21540
ccgctggacg agggcaaccc aacacctagc ctaaagcccg taacactgca gcaggtgctg 21600
cccgcgcttg caccgtccga agaaaagcgc ggcctaaagc gcgagtctgg tgacttggca 21660
cccaccgtgc agctgatggt acccaagcgc cagcgactgg aagatgtctt ggaaaaaatg 21720
accgtggaac ctgggctgga gcccgaggtc cgcgtgcggc caatcaagca ggtggcgccg 21780
ggactgggcg tgcagaccgt ggacgttcag atacccacta ccagtagcac cagtattgcc 21840
accgccacag agggcatgga gacacaaacg tccccggttg cctcagcggt ggcggatgcc 21900
gcggtgcagg cggtcgctgc ggccgcgtcc aagacctcta cggaggtgca aacggacccg 21960
tggatgtttc gcgtttcagc cccccggcgc ccgcgcggtt cgaggaagta cggcgccgcc 22020
agcgcgctac tgcccgaata tgccctacat ccttccattg cgcctacccc cggctatcgt 22080
ggctacacct accgccccag aagacgagca actacccgac gccgaaccac cactggaacc 22140
cgccgccgcc gtcgccgtcg ccagcccgtg ctggccccga tttccgtgcg cagggtggct 22200
cgcgaaggag gcaggaccct ggtgctgcca acagcgcgct accaccccag catcgtttaa 22260
aagccggtct ttgtggttct tgcagatatg gccctcacct gccgcctccg tttcccggtg 22320
ccgggattcc gaggaagaat gcaccgtagg aggggcatgg ccggccacgg cctgacgggc 22380
ggcatgcgtc gtgcgcacca ccggcggcgg cgcgcgtcgc accgtcgcat gcgcggcggt 22440
atcctgcccc tccttattcc actgatcgcc gcggcgattg gcgccgtgcc cggaattgca 22500
tccgtggcct tgcaggcgca gagacactga ttaaaaacaa gttgcatgtg gaaaaatcaa 22560
aataaaaagt ctggactctc acgctcgctt ggtcctgtaa ctattttgta gaatggaaga 22620
catcaacttt gcgtctctgg ccccgcgaca cggctcgcgc ccgttcatgg gaaactggca 22680
agatatcggc accagcaata tgagcggtgg cgccttcagc tggggctcgc tgtggagcgg 22740
cattaaaaat ttcggttcca ccgttaagaa ctatggcagc aaggcctgga acagcagcac 22800
aggccagatg ctgagggata agttgaaaga gcaaaatttc caacaaaagg tggtagatgg 22860
cctggcctct ggcattagcg gggtggtgga cctggccaac caggcagtgc aaaataagat 22920
taacagtaag cttgatcccc gccctcccgt agaggagcct ccaccggccg tggagacagt 22980
gtctccagag gggcgtggcg aaaagcgtcc gcgccccgac agggaagaaa ctctggtgac 23040
gcaaatagac gagcctccct cgtacgagga ggcactaaag caaggcctgc ccaccacccg 23100
tcccatcgcg cccatggcta ccggagtgct gggccagcac acacccgtaa cgctggacct 23160
gcctcccccc gccgacaccc agcagaaacc tgtgctgcca ggcccgaccg ccgttgttgt 23220
aacccgtcct agccgcgcgt ccctgcgccg cgccgccagc ggtccgcgat cgttgcggcc 23280
cgtagccagt ggcaactggc aaagcacact gaacagcatc gtgggtctgg gggtgcaatc 23340
cctgaagcgc cgacgatgct tctgaatagc taacgtgtcg tatgtgtgtc atgtatgcgt 23400
ccatgtcgcc gccagaggag ctgctgagcc gccgcgcgcc cgctttccaa gatggctacc 23460
ccttcgatga tgccgcagtg gtcttacatg cacatctcgg gccaggacgc ctcggagtac 23520
ctgagccccg ggctggtgca gtttgcccgc gccaccgaga cgtacttcag cctgaataac 23580
aagtttagaa accccacggt ggcgcctacg cacgacgtga ccacagaccg gtcccagcgt 23640
ttgacgctgc ggttcatccc tgtggaccgt gaggatactg cgtactcgta caaggcgcgg 23700
ttcaccctag ctgtgggtga taaccgtgtg ctggacatgg cttccacgta ctttgacatc 23760
cgcggcgtgc tggacagggg ccctactttt aagccctact ctggcactgc ctacaacgcc 23820
ctggctccca agggtgcccc aaatccttgc gaatgggatg aagctgctac tgctcttgaa 23880
ataaacctag aagaagagga cgatgacaac gaagacgaag tagacgagca agctgagcag 23940
caaaaaactc acgtatttgg gcaggcgcct tattctggta taaatattac aaaggagggt 24000
attcaaatag gtgtcgaagg tcaaacacct aaatatgccg ataaaacatt tcaacctgaa 24060
cctcaaatag gagaatctca gtggtacgaa actgaaatta atcatgcagc tgggagagtc 24120
cttaaaaaga ctaccccaat gaaaccatgt tacggttcat atgcaaaacc cacaaatgaa 24180
aatggagggc aaggcattct tgtaaagcaa caaaatggaa agctagaaag tcaagtggaa 24240
atgcaatttt tctcaactac tgaggcgacc gcaggcaatg gtgataactt gactcctaaa 24300
gtggtattgt acagtgaaga tgtagatata gaaaccccag acactcatat ttcttacatg 24360
cccactatta aggaaggtaa ctcacgagaa ctaatgggcc aacaatctat gcccaacagg 24420
cctaattaca ttgcttttag ggacaatttt attggtctaa tgtattacaa cagcacgggt 24480
aatatgggtg ttctggcggg ccaagcatcg cagttgaatg ctgttgtaga tttgcaagac 24540
agaaacacag agctttcata ccagcttttg cttgattcca ttggtgatag aaccaggtac 24600
ttttctatgt ggaatcaggc tgttgacagc tatgatccag atgttagaat tattgaaaat 24660
catggaactg aagatgaact tccaaattac tgctttccac tgggaggtgt gattaataca 24720
gagactctta ccaaggtaaa acctaaaaca ggtcaggaaa atggatggga aaaagatgct 24780
acagaatttt cagataaaaa tgaaataaga gttggaaata attttgccat ggaaatcaat 24840
ctaaatgcca acctgtggag aaatttcctg tactccaaca tagcgctgta tttgcccgac 24900
aagctaaagt acagtccttc caacgtaaaa atttctgata acccaaacac ctacgactac 24960
atgaacaagc gagtggtggc tcccgggtta gtggactgct acattaacct tggagcacgc 25020
tggtcccttg actatatgga caacgtcaac ccatttaacc accaccgcaa tgctggcctg 25080
cgctaccgct caatgttgct gggcaatggt cgctatgtgc ccttccacat ccaggtgcct 25140
cagaagttct ttgccattaa aaacctcctt ctcctgccgg gctcatacac ctacgagtgg 25200
aacttcagga aggatgttaa catggttctg cagagctccc taggaaatga cctaagggtt 25260
gacggagcca gcattaagtt tgatagcatt tgcctttacg ccaccttctt ccccatggcc 25320
cacaacaccg cctccacgct tgaggccatg cttagaaacg acaccaacga ccagtccttt 25380
aacgactatc tctccgccgc caacatgctc taccctatac ccgccaacgc taccaacgtg 25440
cccatatcca tcccctcccg caactgggcg gctttccgcg gctgggcctt cacgcgcctt 25500
aagactaagg aaaccccatc actgggctcg ggctacgacc cttattacac ctactctggc 25560
tctataccct acctagatgg aaccttttac ctcaaccaca cctttaagaa ggtggccatt 25620
acctttgact cttctgtcag ctggcctggc aatgaccgcc tgcttacccc caacgagttt 25680
gaaattaagc gctcagttga cggggagggt tacaacgttg cccagtgtaa catgaccaaa 25740
gactggttcc tggtacaaat gctagctaac tacaacattg gctaccaggg cttctatatc 25800
ccagagagct acaaggaccg catgtactcc ttctttagaa acttccagcc catgagccgt 25860
caggtggtgg atgatactaa atacaaggac taccaacagg tgggcatcct acaccaacac 25920
aacaactctg gatttgttgg ctaccttgcc cccaccatgc gcgaaggaca ggcctaccct 25980
gctaacttcc cctatccgct tataggcaag accgcagttg acagcattac ccagaaaaag 26040
tttctttgcg atcgcaccct ttggcgcatc ccattctcca gtaactttat gtccatgggc 26100
gcactcacag acctgggcca aaaccttctc tacgccaact ccgcccacgc gctagacatg 26160
acttttgagg tggatcccat ggacgagccc acccttcttt atgttttgtt tgaagtcttt 26220
gacgtggtcc gtgtgcaccg gccgcaccgc ggcgtcatcg aaaccgtgta cctgcgcacg 26280
cccttctcgg ccggcaacgc cacaacataa agaagcaagc aacatcaaca acagctgccg 26340
ccatgggctc cagtgagcag gaactgaaag ccattgtcaa agatcttggt tgtgggccat 26400
attttttggg cacctatgac aagcgctttc caggctttgt ttctccacac aagctcgcct 26460
gcgccatagt caatacggcc ggtcgcgaga ctgggggcgt acactggatg gcctttgcct 26520
ggaacccgca ctcaaaaaca tgctacctct ttgagccctt tggcttttct gaccagcgac 26580
tcaagcaggt ttaccagttt gagtacgagt cactcctgcg ccgtagcgcc attgcttctt 26640
cccccgaccg ctgtataacg ctggaaaagt ccacccaaag cgtacagggg cccaactcgg 26700
ccgcctgtgg actattctgc tgcatgtttc tccacgcctt tgccaactgg ccccaaactc 26760
ccatggatca caaccccacc atgaacctta ttaccggggt acccaactcc atgctcaaca 26820
gtccccaggt acagcccacc ctgcgtcgca accaggaaca gctctacagc ttcctggagc 26880
gccactcgcc ctacttccgc agccacagtg cgcagattag gagcgccact tctttttgtc 26940
acttgaaaaa catgtaaaaa taatgtacta gagacacttt caataaaggc aaatgctttt 27000
atttgtacac tctcgggtga ttatttaccc ccacccttgc cgtctgcgcc gtttaaaaat 27060
caaaggggtt ctgccgcgca tcgctatgcg ccactggcag ggacacgttg cgatactggt 27120
gtttagtgct ccacttaaac tcaggcacaa ccatccgcgg cagctcggtg aagttttcac 27180
tccacaggct gcgcaccatc accaacgcgt ttagcaggtc gggcgccgat atcttgaagt 27240
cgcagttggg gcctccgccc tgcgcgcgcg agttgcgata cacagggttg cagcactgga 27300
acactatcag cgccgggtgg tgcacgctgg ccagcacgct cttgtcggag atcagatccg 27360
cgtccaggtc ctccgcgttg ctcagggcga acggagtcaa ctttggtagc tgccttccca 27420
aaaagggcgc gtgcccaggc tttgagttgc actcgcaccg tagtggcatc aaaaggtgac 27480
cgtgcccggt ctgggcgtta ggatacagcg cctgcataaa agccttgatc tgcttaaaag 27540
ccacctgagc ctttgcgcct tcagagaaga acatgccgca agacttgccg gaaaactgat 27600
tggccggaca ggccgcgtcg tgcacgcagc accttgcgtc ggtgttggag atctgcacca 27660
catttcggcc ccaccggttc ttcacgatct tggccttgct agactgctcc ttcagcgcgc 27720
gctgcccgtt ttcgctcgtc acatccattt caatcacgtg ctccttattt atcataatgc 27780
ttccgtgtag acacttaagc tcgccttcga tctcagcgca gcggtgcagc cacaacgcgc 27840
agcccgtggg ctcgtgatgc ttgtaggtca cctctgcaaa cgactgcagg tacgcctgca 27900
ggaatcgccc catcatcgtc acaaaggtct tgttgctggt gaaggtcagc tgcaacccgc 27960
ggtgctcctc gttcagccag gtcttgcata cggccgccag agcttccact tggtcaggca 28020
gtagtttgaa gttcgccttt agatcgttat ccacgtggta cttgtccatc agcgcgcgcg 28080
cagcctccat gcccttctcc cacgcagaca cgatcggcac actcagcggg ttcatcaccg 28140
taatttcact ttccgcttcg ctgggctctt cctcttcctc ttgcgtccgc ataccacgcg 28200
ccactgggtc gtcttcattc agccgccgca ctgtgcgctt acctcctttg ccatgcttga 28260
ttagcaccgg tgggttgctg aaacccacca tttgtagcgc cacatcttct ctttcttcct 28320
cgctgtccac gattacctct ggtgatggcg ggcgctcggg cttgggagaa gggcgcttct 28380
ttttcttctt gggcgcaatg gccaaatccg ccgccgaggt cgatggccgc gggctgggtg 28440
tgcgcggcac cagcgcgtct tgtgatgagt cttcctcgtc ctcggactcg atacgccgcc 28500
tcatccgctt ttttgggggc gcccggggag gcggcggcga cggggacggg gacgacacgt 28560
cctccatggt tgggggacgt cgcgccgcac cgcgtccgcg ctcgggggtg gtttcgcgct 28620
gctcctcttc ccgactggcc atttccttct cctataggca gaaaaagatc atggagtcag 28680
tcgagaagaa ggacagccta accgccccct ctgagttcgc caccaccgcc tccaccgatg 28740
ccgccaacgc gcctaccacc ttccccgtcg aggcaccccc gcttgaggag gaggaagtga 28800
ttatcgagca ggacccaggt tttgtaagcg aagacgacga ggaccgctca gtaccaacag 28860
aggataaaaa gcaagaccag gacaacgcag aggcaaacga ggaacaagtc gggcgggggg 28920
acgaaaggca tggcgactac ctagatgtgg gagacgacgt gctgttgaag catctgcagc 28980
gccagtgcgc cattatctgc gacgcgttgc aagagcgcag cgatgtgccc ctcgccatag 29040
cggatgtcag ccttgcctac gaacgccacc tattctcacc gcgcgtaccc cccaaacgcc 29100
aagaaaacgg cacatgcgag cccaacccgc gcctcaactt ctaccccgta tttgccgtgc 29160
cagaggtgct tgccacctat cacatctttt tccaaaactg caagataccc ctatcctgcc 29220
gtgccaaccg cagccgagcg gacaagcagc tggccttgcg gcagggcgct gtcatacctg 29280
atatcgcctc gctcaacgaa gtgccaaaaa tctttgaggg tcttggacgc gacgagaagc 29340
gcgcggcaaa cgctctgcaa caggaaaaca gcgaaaatga aagtcactct ggagtgttgg 29400
tggaactcga gggtgacaac gcgcgcctag ccgtactaaa acgcagcatc gaggtcaccc 29460
actttgccta cccggcactt aacctacccc ccaaggtcat gagcacagtc atgagtgagc 29520
tgatcgtgcg ccgtgcgcag cccctggaga gggatgcaaa tttgcaagaa caaacagagg 29580
agggcctacc cgcagttggc gacgagcagc tagcgcgctg gcttcaaacg cgcgagcctg 29640
ccgacttgga ggagcgacgc aaactaatga tggccgcagt gctcgttacc gtggagcttg 29700
agtgcatgca gcggttcttt gctgacccgg agatgcagcg caagctagag gaaacattgc 29760
actacacctt tcgacagggc tacgtacgcc aggcctgcaa gatctccaac gtggagctct 29820
gcaacctggt ctcctacctt ggaattttgc acgaaaaccg ccttgggcaa aacgtgcttc 29880
attccacgct caagggcgag gcgcgccgcg actacgtccg cgactgcgtt tacttatttc 29940
tatgctacac ctggcagacg gccatgggcg tttggcagca gtgcttggag gagtgcaacc 30000
tcaaggagct gcagaaactg ctaaagcaaa acttgaagga cctatggacg gccttcaacg 30060
agcgctccgt ggccgcgcac ctggcggaca tcattttccc cgaacgcctg cttaaaaccc 30120
tgcaacaggg tctgccagac ttcaccagtc aaagcatgtt gcagaacttt aggaacttta 30180
tcctagagcg ctcaggaatc ttgcccgcca cctgctgtgc acttcctagc gactttgtgc 30240
ccattaagta ccgcgaatgc cctccgccgc tttggggcca ctgctacctt ctgcagctag 30300
ccaactacct tgcctaccac tctgacataa tggaagacgt gagcggtgac ggtctactgg 30360
agtgtcactg tcgctgcaac ctatgcaccc cgcaccgctc cctggtttgc aattcgcagc 30420
tgcttaacga aagtcaaatt atcggtacct ttgagctgca gggtccctcg cctgacgaaa 30480
agtccgcggc tccggggttg aaactcactc cggggctgtg gacgtcggct taccttcgca 30540
aatttgtacc tgaggactac cacgcccacg agattaggtt ctacgaagac caatcccgcc 30600
cgccaaatgc ggagcttacc gcctgcgtca ttacccaggg ccacattctt ggccaattgc 30660
aagccatcaa caaagcccgc caagagtttc tgctacgaaa gggacggggg gtttacttgg 30720
acccccagtc cggcgaggag ctcaacccaa tccccccgcc gccgcagccc tatcagcagc 30780
agccgcgggc ccttgcttcc caggatggca cccaaaaaga agctgcagct gccgccgcca 30840
cccacggacg aggaggaata ctgggacagt caggcagagg aggttttgga cgaggaggag 30900
gaggacatga tggaagactg ggagagccta gacgaggaag cttccgaggt cgaagaggtg 30960
tcagacgaaa caccgtcacc ctcggtcgca ttcccctcgc cggcgcccca gaaatcggca 31020
accggttcca gcatggctac aacctccgct cctcaggcgc cgccggcact gcccgttcgc 31080
cgacccaacc gtagatggga caccactgga accagggccg gtaagtccaa gcagccgccg 31140
ccgttagccc aagagcaaca acagcgccaa ggctaccgct catggcgcgg gcacaagaac 31200
gccatagttg cttgcttgca agactgtggg ggcaacatct ccttcgcccg ccgctttctt 31260
ctctaccatc acggcgtggc cttcccccgt aacatcctgc attactaccg tcatctctac 31320
agcccatact gcaccggcgg cagcggcagc ggcagcaaca gcagcggcca cacagaagca 31380
aaggcgaccg gatagcaaga ctctgacaaa gcccaagaaa tccacagcgg cggcagcagc 31440
aggaggagga gcgctgcgtc tggcgcccaa cgaacccgta tcgacccgcg agcttagaaa 31500
caggattttt cccactctgt atgctatatt tcaacagagc aggggccaag aacaagagct 31560
gaaaataaaa aacaggtctc tgcgatccct cacccgcagc tgcctgtatc acaaaagcga 31620
agatcagctt cggcgcacgc tggaagacgc ggaggctctc ttcagtaaat actgcgcgct 31680
gactcttaag gactagtttc gcgccctttc tcaaatttaa gcgcgaaaac tacgtcatct 31740
ccagcggcca cacccggcgc cagcacctgt cgtcagcgcc attatgagca aggaaattcc 31800
cacgccctac atgtggagtt accagccaca aatgggactt gcggctggag ctgcccaaga 31860
ctactcaacc cgaataaact acatgagcgc gggaccccac atgatatccc gggtcaacgg 31920
aatccgcgcc caccgaaacc gaattctctt ggaacaggcg gctattacca ccacacctcg 31980
taataacctt aatccccgta gttggcccgc tgccctggtg taccaggaaa gtcccgctcc 32040
caccactgtg gtacttccca gagacgccca ggccgaagtt cagatgacta actcaggggc 32100
gcagcttgcg ggcggctttc gtcacagggt gcggtcgccc gggcagggta taactcacct 32160
gacaatcaga gggcgaggta ttcagctcaa cgacgagtcg gtgagctcct cgcttggtct 32220
ccgtccggac gggacatttc agatcggcgg cgccggccgt ccttcattca cgcctcgtca 32280
ggcaatccta actctgcaga cctcgtcctc tgagccgcgc tctggaggca ttggaactct 32340
gcaatttatt gaggagtttg tgccatcggt ctactttaac cccttctcgg gacctcccgg 32400
ccactatccg gatcaattta ttcctaactt tgacgcggta aaggactcgg cggacggcta 32460
cgactgaatg ttaagtggag aggcagagca actgcgcctg aaacacctgg tccactgtcg 32520
ccgccacaag tgctttgccc gcgactccgg tgagttttgc tactttgaat tgcccgagga 32580
tcatatcgag ggcccggcgc acggcgtccg gcttaccgcc cagggagagc ttgcccgtag 32640
cctgattcgg gagtttaccc agcgccccct gctagttgag cgggacaggg gaccctgtgt 32700
tctcactgtg atttgcaact gtcctaacct tggattacat caagatcctc tagttataac 32760
tagagtaccc ggggatctta ttccctttaa ctaataaaaa aaaataataa agcatcactt 32820
acttaaaatc agttagcaaa tttctgtcca gtttattcag cagcacctcc ttgccctcct 32880
cccagctctg gtattgcagc ttcctcctgg ctgcaaactt tctccacaat ctaaatggaa 32940
tgtcagtttc ctcctgttcc tgtccatccg cacccactat cttcatgttg ttgcagatga 33000
agcgcgcaag accgtctgaa gataccttca accccgtgta tccatatgac acggaaaccg 33060
gtcctccaac tgtgcctttt cttactcctc cctttgtatc ccccaatggg tttcaagaga 33120
gtccccctgg ggtactctct ttgcgcctat ccgaacctct agttacctcc aatggcatgc 33180
ttgcgctcaa aatgggcaac ggcctctctc tggacgaggc cggcaacctt acctcccaaa 33240
atgtaaccac tgtgagccca cctctcaaaa aaaccaagtc aaacataaac ctggaaatat 33300
ctgcacccct cacagttacc tcagaagccc taactgtggc tgccgccgca cctctaatgg 33360
tcgcgggcaa cacactcacc atgcaatcac aggccccgct aaccgtgcac gactccaaac 33420
ttagcattgc cacccaagga cccctcacag tgtcagaagg aaagctagcc ctgcaaacat 33480
caggccccct caccaccacc gatagcagta cccttactat cactgcctca ccccctctaa 33540
ctactgccac tggtagcttg ggcattgact tgaaagagcc catttataca caaaatggaa 33600
aactaggact aaagtacggg gctcctttgc atgtaacaga cgacctaaac actttgaccg 33660
tagcaactgg tccaggtgtg actattaata atacttcctt gcaaactaaa gttactggag 33720
ccttgggttt tgattcacaa ggcaatatgc aacttaatgt agcaggagga ctaaggattg 33780
attctcaaaa cagacgcctt atacttgatg ttagttatcc gtttgatgct caaaaccaac 33840
taaatctaag actaggacag ggccctcttt ttataaactc agcccacaac ttggatatta 33900
actacaacaa aggcctttac ttgtttacag cttcaaacaa ttccaaaaag cttgaggtta 33960
acctaagcac tgccaagggg ttgatgtttg acgctacagc catagccatt aatgcaggag 34020
atgggcttga atttggttca cctaatgcac caaacacaaa tcccctcaaa acaaaaattg 34080
gccatggcct agaatttgat tcaaacaagg ctatggttcc taaactagga actggcctta 34140
gttttgacag cacaggtgcc attacagtag gaaacaaaaa taatgataag ctaactttgt 34200
ggaccacacc agctccatct cctaactgta gactaaatgc agagaaagat gctaaactca 34260
ctttggtctt aacaaaatgt ggcagtcaaa tacttgctac agtttcagtt ttggctgtta 34320
aaggcagttt ggctccaata tctggaacag ttcaaagtgc tcatcttatt ataagatttg 34380
acgaaaatgg agtgctacta aacaattcct tcctggaccc agaatattgg aactttagaa 34440
atggagatct tactgaaggc acagcctata caaacgctgt tggatttatg cctaacctat 34500
cagcttatcc aaaatctcac ggtaaaactg ccaaaagtaa cattgtcagt caagtttact 34560
taaacggaga caaaactaaa cctgtaacac taaccattac actaaacggt acacaggaaa 34620
caggagacac aactccaagt gcatactcta tgtcattttc atgggactgg tctggccaca 34680
actacattaa tgaaatattt gccacatcct cttacacttt ttcatacatt gcccaagaat 34740
aaagaatcgt ttgtgttatg tttcaacgtg tttatttttc aattgcagaa aatttcaagt 34800
catttttcat tcagtagtat agccccacca ccacatagct tatacagatc accgtacctt 34860
aatcaaactc acagaaccct agtattcaac ctgccacctc cctcccaaca cacagagtac 34920
acagtccttt ctccccggct ggccttaaaa agcatcatat catgggtaac agacatattc 34980
ttaggtgtta tattccacac ggtttcctgt cgagccaaac gctcatcagt gatattaata 35040
aactccccgg gcagctcact taagttcatg tcgctgtcca gctgctgagc cacaggctgc 35100
tgtccaactt gcggttgctt aacgggcggc gaaggagaag tccacgccta catgggggta 35160
gagtcataat cgtgcatcag gatagggcgg tggtgctgca gcagcgcgcg aataaactgc 35220
tgccgccgcc gctccgtcct gcaggaatac aacatggcag tggtctcctc agcgatgatt 35280
cgcaccgccc gcagcataag gcgccttgtc ctccgggcac agcagcgcac cctgatctca 35340
cttaaatcag cacagtaact gcagcacagc accacaatat tgttcaaaat cccacagtgc 35400
aaggcgctgt atccaaagct catggcgggg accacagaac ccacgtggcc atcataccac 35460
aagcgcaggt agattaagtg gcgacccctc ataaacacgc tggacataaa cattacctct 35520
tttggcatgt tgtaattcac cacctcccgg taccatataa acctctgatt aaacatggcg 35580
ccatccacca ccatcctaaa ccagctggcc aaaacctgcc cgccggctat acactgcagg 35640
gaaccgggac tggaacaatg acagtggaga gcccaggact cgtaaccatg gatcatcatg 35700
ctcgtcatga tatcaatgtt ggcacaacac aggcacacgt gcatacactt cctcaggatt 35760
acaagctcct cccgcgttag aaccatatcc cagggaacaa cccattcctg aatcagcgta 35820
aatcccacac tgcagggaag acctcgcacg taactcacgt tgtgcattgt caaagtgtta 35880
cattcgggca gcagcggatg atcctccagt atggtagcgc gggtttctgt ctcaaaagga 35940
ggtagacgat ccctactgta cggagtgcgc cgagacaacc gagatcgtgt tggtcgtagt 36000
gtcatgccaa atggaacgcc ggacgtagtc atatttcctg aagcaaaacc aggtgcgggc 36060
gtgacaaaca gatctgcgtc tccggtctcg ccgcttagat cgctctgtgt agtagttgta 36120
gtatatccac tctctcaaag catccaggcg ccccctggct tcgggttcta tgtaaactcc 36180
ttcatgcgcc gctgccctga taacatccac caccgcagaa taagccacac ccagccaacc 36240
tacacattcg ttctgcgagt cacacacggg aggagcggga agagctggaa gaaccatgtt 36300
ttttttttta ttccaaaaga ttatccaaaa cctcaaaatg aagatctatt aagtgaacgc 36360
gctcccctcc ggtggcgtgg tcaaactcta cagccaaaga acagataatg gcatttgtaa 36420
gatgttgcac aatggcttcc aaaaggcaaa cggccctcac gtccaagtgg acgtaaaggc 36480
taaacccttc agggtgaatc tcctctataa acattccagc accttcaacc atgcccaaat 36540
aattctcatc tcgccacctt ctcaatatat ctctaagcaa atcccgaata ttaagtccgg 36600
ccattgtaaa aatctgctcc agagcgccct ccaccttcag cctcaagcag cgaatcatga 36660
ttgcaaaaat tcaggttcct cacagacctg tataagattc aaaagcggaa cattaacaaa 36720
aataccgcga tcccgtaggt cccttcgcag ggccagctga acataatcgt gcaggtctgc 36780
acggaccagc gcggccactt ccccgccagg aaccttgaca aaagaaccca cactgattat 36840
gacacgcata ctcggagcta tgctaaccag cgtagccccg atgtaagctt tgttgcatgg 36900
gcggcgatat aaaatgcaag gtgctgctca aaaaatcagg caaagcctcg cgcaaaaaag 36960
aaagcacatc gtagtcatgc tcatgcagat aaaggcaggt aagctccgga accaccacag 37020
aaaaagacac catttttctc tcaaacatgt ctgcgggttt ctgcataaac acaaaataaa 37080
ataacaaaaa aacatttaaa cattagaagc ctgtcttaca acaggaaaaa caacccttat 37140
aagcataaga cggactacgg ccatgccggc gtgaccgtaa aaaaactggt caccgtgatt 37200
aaaaagcacc accgacagct cctcggtcat gtccggagtc ataatgtaag actcggtaaa 37260
cacatcaggt tgattcatcg gtcagtgcta aaaagcgacc gaaatagccc gggggaatac 37320
atacccgcag gcgtagagac aacattacag cccccatagg aggtataaca aaattaatag 37380
gagagaaaaa cacataaaca cctgaaaaac cctcctgcct aggcaaaata gcaccctccc 37440
gctccagaac aacatacagc gcttcacagc ggcagcctaa cagtcagcct taccagtaaa 37500
aaagaaaacc tattaaaaaa acaccactcg acacggcacc agctcaatca gtcacagtgt 37560
aaaaaagggc caagtgcaga gcgagtatat ataggactaa aaaatgacgt aacggttaaa 37620
gtccacaaaa aacacccaga aaaccgcacg cgaacctacg cccagaaacg aaagccaaaa 37680
aacccacaac ttcctcaaat cgtcacttcc gttttcccac gttacgtaac ttcccatttt 37740
aagaaaacta caattcccaa cacatacaag ttactccgcc ctaaaaccta cgtcacccgc 37800
cccgttccca cgccccgcgc cacgtcacaa actccacccc ctcattatca tattggcttc 37860
aatccaaaat aaggtatatt attgatgata ct 37892
<211> 24
<212> DNA
<213> Artificial(人工序列)
aatcggaaag cggacgcgga attt 24
<211> 24
<212> DNA
<213> Artificial(人工序列)
aagcgctcac gggatactcg attt 24
<211> 55
<212> DNA
<213> Artificial(人工序列)
aatcggaaag cggacgcgga atttaccacc atggagagcg acgacagcgg ccatg 55
<211> 46
<212> DNA
<213> Artificial(人工序列)
aagcgctcac gggatactcg atttagcgag atccggtgga gccggg 46
Claims (9)
1.一种腺病毒质粒载体,包含5型腺病毒骨架基因序列,两个反向末端重复序列,启动子序列,多聚A尾序列,其特征在于:所述腺病毒质粒载体的启动子序列和多聚A尾序列之间有如SEQ NO.1所示的位点序列1及如SEQ NO.2所示的位点序列2,位点序列1及位点序列2中由一个唯一的平末端限制性核酸内切酶识别位点序列连接。
2.如权利要求1所述的腺病毒质粒载体,其特征还在于:所述位点序列1及位点序列2之间由一个SwaI限制性内切酶识别序列连接,连接后的序列如SEQ No.3所示。
3.如权利要求1所述的腺病毒质粒载体,其特征还在于:包含绿色荧光蛋白或红色荧光蛋白的开放阅读框序列,所述开放阅读框序列位于另一启动子序列下游,用于编码绿色荧光蛋白或红色荧光蛋白。
4.如权利要求1或2所述的腺病毒质粒载体,其特征还在于:所述位点序列1与位点序列2之间有细菌自杀基因序列。
5.如权利要求4所述的腺病毒质粒载体,其特征还在于:所述细菌自杀基因序列为SEQNO.4所示ccdB表达盒,表达盒两端有SwaI酶切位点,所述的腺病毒质粒载体序列为SEQNO.5。
6.如权利要求5所述的腺病毒质粒载体,其特征在于:所述腺病毒质粒载体包含绿色荧光蛋白开放阅读框序列,序列为SEQ NO.6。
7.如权利要求5所述的腺病毒质粒载体,其特征在于:所述腺病毒质粒载体包含红色荧光蛋白开放阅读框序列,序列为SEQ NO.7。
8.一种一步法构建腺病毒的方法,包含以下步骤:
步骤1:设计5’端包含SEQ NO.1的上游引物及5’端包含SEQ NO.2的下游引物,PCR扩增获得目的基因;
步骤2:用限制性内切酶酶切位点序列1及位点序列2之间的识别位点序列,获得线性化载体片段;
步骤2:纯化后的目的基因片段与线性化载体片段混合,通过Gibson DNA组装,获得带有目的基因的重组腺病毒质粒;
步骤3:将带有目的基因的重组腺病毒质粒经PacI酶切线性化,转染到HEK-293或293A或293T腺病毒包装细胞中,5-7天收集细胞,裂解细胞获得腺病毒初提液;
步骤3:腺病毒初提液感染HEK-293或293A或293T细胞,进一步扩增,获得高滴度病毒液。
9.一种腺病毒试剂盒,其特征在于:包含权利要求1-3或5-7所述腺病毒质粒载体。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111471784.2A CN114085871A (zh) | 2021-12-02 | 2021-12-02 | 一种腺病毒质粒载体及腺病毒的制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111471784.2A CN114085871A (zh) | 2021-12-02 | 2021-12-02 | 一种腺病毒质粒载体及腺病毒的制备方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114085871A true CN114085871A (zh) | 2022-02-25 |
Family
ID=80306477
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111471784.2A Withdrawn CN114085871A (zh) | 2021-12-02 | 2021-12-02 | 一种腺病毒质粒载体及腺病毒的制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114085871A (zh) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2001252941A1 (en) * | 2000-03-21 | 2001-10-03 | The University Of Virginia Patent Foundation | Osteocalcin promoter directed adenovirus replicaton for therapy |
CN101319228A (zh) * | 2008-07-16 | 2008-12-10 | 深圳市第二人民医院 | 一种含人bmp重组腺病毒载体及其构建方法 |
CN101503677A (zh) * | 2008-12-26 | 2009-08-12 | 四川大学华西医院 | Survivin启动子调控人NIS基因的重组腺病毒及其构建方法和应用 |
CN102168075A (zh) * | 2011-02-28 | 2011-08-31 | 重庆医科大学附属儿童医院 | 一种携带大鼠RARγ基因的重组腺病毒及构建方法 |
CN108841866A (zh) * | 2018-06-04 | 2018-11-20 | 云舟生物科技(广州)有限公司 | 一种腺病毒载体及其构建方法 |
CN109295096A (zh) * | 2018-10-29 | 2019-02-01 | 中国疾病预防控制中心病毒病预防控制所 | 腺病毒载体系统及重组腺病毒构建方法 |
CN111471715A (zh) * | 2020-04-07 | 2020-07-31 | 南昌大学第一附属医院 | 一种腺病毒载体及其构建方法和用途 |
-
2021
- 2021-12-02 CN CN202111471784.2A patent/CN114085871A/zh not_active Withdrawn
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2001252941A1 (en) * | 2000-03-21 | 2001-10-03 | The University Of Virginia Patent Foundation | Osteocalcin promoter directed adenovirus replicaton for therapy |
CN101319228A (zh) * | 2008-07-16 | 2008-12-10 | 深圳市第二人民医院 | 一种含人bmp重组腺病毒载体及其构建方法 |
CN101503677A (zh) * | 2008-12-26 | 2009-08-12 | 四川大学华西医院 | Survivin启动子调控人NIS基因的重组腺病毒及其构建方法和应用 |
CN102168075A (zh) * | 2011-02-28 | 2011-08-31 | 重庆医科大学附属儿童医院 | 一种携带大鼠RARγ基因的重组腺病毒及构建方法 |
CN108841866A (zh) * | 2018-06-04 | 2018-11-20 | 云舟生物科技(广州)有限公司 | 一种腺病毒载体及其构建方法 |
CN109295096A (zh) * | 2018-10-29 | 2019-02-01 | 中国疾病预防控制中心病毒病预防控制所 | 腺病毒载体系统及重组腺病毒构建方法 |
CN111471715A (zh) * | 2020-04-07 | 2020-07-31 | 南昌大学第一附属医院 | 一种腺病毒载体及其构建方法和用途 |
Non-Patent Citations (4)
Title |
---|
BI YOUKUN等: "A Novel System for Simple Rapid Adenoviral Vector Construction to Facilitate CRISPR/Cas9-Mediated Genome Editing" * |
NI NA等: "A one-step construction of adenovirus (OSCA) system using the Gibson DNA Assembly technology" * |
程志安 等: "重组腺病毒载体Ad-LMP-1的构建及其转染MSC后成骨活性" * |
黄蕾 等: "利用DNA Assembly方法构建重组腺病毒载体" * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR102111244B1 (ko) | 이종 유전자의 발현을 위한 강력한 짧은 프로모터 | |
KR100987360B1 (ko) | 원숭이 아데노바이러스 핵산과 아미노산 서열, 이를함유하는 벡터 및 사용방법 | |
RU2731342C1 (ru) | Фармацевтическое средство и способ его использования для индукции специфического иммунитета против вируса тяжелого острого респираторного синдрома SARS-CoV-2 (варианты) | |
KR102502974B1 (ko) | 향상된 입양 세포 치료 | |
US8932607B2 (en) | Batches of recombinant adenovirus with altered terminal ends | |
KR20150014505A (ko) | 아과 e 원숭이 아데노바이러스 a1302, a1320, a1331 및 a1337 및 이것들의 사용 | |
AU2013231423B2 (en) | Batches of recombinant adenovirus with altered terminal ends | |
AU2022203504A1 (en) | Oncolytic tumor viruses and methods of use | |
US20040136963A1 (en) | Simian adenovirus vectors and methods of use | |
JP2015083006A (ja) | サルEアデノウイルスSAdV−39、−25.2、−26、−30、−37および−38 | |
CN101213204A (zh) | 丙型肝炎病毒核酸疫苗 | |
KR20200066349A (ko) | 복제 가능 아데노바이러스 벡터 | |
CN109312362A (zh) | 有效和平衡的双向启动子 | |
EP1606397A1 (en) | Adenovirus serotype 24 vectors, nucleic acids and virus produced thereby | |
WO2004018627A2 (en) | Methods for propagating adenovirus and virus produced thereby | |
CN114085871A (zh) | 一种腺病毒质粒载体及腺病毒的制备方法 | |
RU2821989C1 (ru) | Новый аденовирусный вектор, не включающий компетентный по репликации аденовирус, и его применение | |
KR20220106072A (ko) | 복제가능 아데노바이러스를 포함하지 않는 신규 아데노바이러스 벡터 및 이의 용도 | |
KR20230008069A (ko) | 아데노바이러스 발현 벡터, 및 제조를 위한 방법 및 세포주 | |
KR20230146436A (ko) | 집단의 재접종을 위한 중증 급성 호흡기 증후군 바이러스 sars-cov-2에 대한 특이 면역의 유도를 위한 제제(변형체)의 용도 | |
KR20230092820A (ko) | Sars-cov-2에 대한 특이적 면역을 유도하기 위한 약제학적 제제 | |
KR20220128632A (ko) | 개선된 aav-abcd1 구축물 및 부신백질이영양증 (ald) 및/또는 부신척수신경병증 (amn)의 치료 또는 예방을 위한 용도 | |
CN109154000A (zh) | 有效和平衡的双向启动子 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220225 |