CN114763538B - 溶藻弧菌噬菌体vB_ValS_R15Z及其应用 - Google Patents
溶藻弧菌噬菌体vB_ValS_R15Z及其应用 Download PDFInfo
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- CN114763538B CN114763538B CN202210249036.8A CN202210249036A CN114763538B CN 114763538 B CN114763538 B CN 114763538B CN 202210249036 A CN202210249036 A CN 202210249036A CN 114763538 B CN114763538 B CN 114763538B
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- vibrio alginolyticus
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Abstract
本发明公开了一种溶藻弧菌噬菌体vB_ValS_R15Z及其应用。该溶藻弧菌噬菌体vB_ValS_R15Z的保藏编号为GDMCC No：62170‑B1。本发明提供的溶藻弧菌噬菌体vB_ValS_R15Z是一株新的裂解性病毒，对溶藻弧菌致病株ATCC 17749T具有专一、显著杀灭作用。此外，在该噬菌体基因组中未检测到毒力因子基因和耐药性基因，表明该噬菌体不会增强宿主毒效强度、不会造成周围水体环境的抗药性污染，安全性高、环境友好，因此溶藻弧菌噬菌体vB_ValS_R15Z完全可以用于水产溶藻弧菌病害的防治。
Description
技术领域
本发明涉及微生物应用技术领域，具体涉及一种溶藻弧菌噬菌体vB_ValS_R15Z及其应用。
背景技术
水产病害是水产养殖业最大的威胁之一，其中尤以细菌类病害最为严重，据统计细菌病害约占水产病害一半以上。弧菌(Vibrio)是水产养殖动物的最常见的病原菌之一，迄今为止被证实对水产动物具有致病性的弧菌具有20多种，危害对象包括鱼类、贝类、甲壳类等几乎所有水产动物，给我国的水产养殖事业造成了巨大的损失。溶藻弧菌(Vibrioalginolyticus) 又是弧菌中较为常见的一种致病菌，其数量居海水类弧菌之首，溶藻弧菌会产生诸如溶血毒素、黏附素、耐热性肠毒素以及河鲀毒素等危害水产动物甚至人类的多种毒素。溶藻弧菌感染会使水产动物消化道、皮肤等器官发生病变，引起水产动物伤口感染、腐皮病、败血症等多种疾病，其发病性高、爆发面积广，严重危害水产养殖的健康发展。此外，误食感染溶藻弧菌而未充分煮熟灭菌的水产品，还可能会对人体产生感染，使人体产生腹泻、眩晕等症状。
目前，针对水产病害的防治方法主要有臭氧、紫外线灯物理灭菌法、免疫疫苗法以及消毒剂、抗生素化学法等，前两种方法虽然对生态环境相对友好，但是物理法存在不能覆盖全面水体达不到充分灭菌效果，而免疫法尚未完全成熟，针对性的疫苗种类十分有限，难以满足复杂多样的水产病害需求。所以，目前消毒剂、抗生素化学法依然是防治水产病害最普遍的方法，但由于长期过量、混滥使用已严重污染水产环境并诱发滋生大量抗药致病菌株，反而使得水产病害反复爆发难以根治、日趋严重。同时由于化学药物在水产品中的残留积累，通过食物链传递往往无形中对食用者/人类产生危害，所以抗生素等化学药物防治方法并不符合目前的生态养殖趋势。另外，抗生素防治还存在新抗生素研发周期漫长、研发成本高等问题，随着抗生素药物滥用、水产致病菌耐药菌株的产生，不久的将来水产养殖业可能面临无抗生素可用的窘境，因此寻找高效、环保的替代防治方法迫在眉睫。
噬菌体作为一种可以专一对细菌具有强大杀灭能力的病毒，由于其在自然界中广泛生存分布、对宿主裂解作用高度专一，即整体上具有安全性的特点，当下利用噬菌体防治水产病害问题而备受关注。噬菌体防治主要是利用烈性噬菌体对细菌的裂解作用杀灭致病细菌，以达到治疗和防御水产动物细菌性病害的效果。一种噬菌体通常只会侵染裂解特定的一种或一类宿主菌，不会破坏水产品体内或水产环境中其他菌群结构，且对水产动物细胞无伤害。烈性噬菌体侵入宿主后迅速完成大量的自我增殖复制，进行指数级生长直至宿主裂解，再去侵染裂解其他宿主菌，以此反复，可以迅速有效地杀灭致病菌。此外，噬菌体治疗还有很难产生抗性、无药物残留、研发周期短、成本低等优势。总体而言，噬菌体防治法不仅对水产病害细菌有良好杀灭作用而且对环境友好，所以该方法完全可以替代抗生素用于防控细菌类病害乃至耐药致病菌株病害。
为开发能够安全有效防治溶藻弧菌类水产病害的噬菌体防治方法，首先我们必须筛选出能够安全有效且特异性杀灭溶藻弧菌的噬菌体方可加以利用。但值得注意的是，养殖水体致病菌种类多样，而且同种类不同株系致病性很大程度上也存在差异。噬菌体裂解细菌的作用具有严格的宿主特异性，一般仅限于细菌分类学上“种”的界限之内，大多噬菌体仅对某种细菌里的某些特定株系有裂解作用。基于噬菌体这种严格特异性的特点，要实现噬菌体在水产养殖中的应用，就需要通过不断地筛选以积累针对不同致病菌株、具有裂解作用的噬菌体，建立并扩大涵盖各种致病菌种、致病菌株的噬菌体库以及时应对不同的水产病害。
发明内容
基于噬菌体于自然界中存在的普遍性及其自身对宿主菌的高度专一性，针对溶藻弧菌病害特异筛选出可抑制、杀灭致病菌的噬菌体，以用于防治由溶藻弧菌感染所导致的水产病害。
针对上述目的，本发明采用如下技术方案实现：
本发明的第一个目的是提供一种溶藻弧菌噬菌体vB_ValS_R15Z，其保藏编号为：GDMCC No:62170-B1。
本发明从福建厦门第八市场废水中筛选分离得到一株烈性溶藻弧菌噬菌体，通过形态观察、全基因测序和系统发育分析确定了该噬菌体的形态及分类地位，并将其命名为Vibrio phage vB_ValS_R15Z，于2021年12月30日保藏于广东省微生物菌种保藏中心(GDMCC)，保藏编号为GDMCC No：62170-B1，保藏地址为：广州市先烈中路100号大院59号楼5楼，邮编：510070，分类命名：Vibrio phage。
进一步地，透射电子显微镜观察显示，溶藻弧菌噬菌体vB_ValS_R15Z头部长69nm、宽55nm，尾部长16nm，属于短尾类病毒，结合其系统发育树分析，根据国际病毒分类委员会(ICTV)最新病毒分类系统的标准，确定溶藻弧菌噬菌体vB_ValS_R15Z隶属于Caudovirales(目)、Autographiviridae(科)、Colwellvirinae(亚科)、Kaohsiungvirus(属)。
进一步地，所述溶藻弧菌噬菌体vB_ValS_R15Z对溶藻弧菌ATCC 17749T(Vibrioalginolyticus ATCC 17749T)高度专一性裂解作用。
进一步地，将溶藻弧菌噬菌体vB_ValS_R15Z基因组分别在致病菌毒力因子数据库(VFDB)以及综合耐药性数据库(CARD)中进行对比分析，未检测到毒力基因与抗药性基因，表明溶藻弧菌噬菌体vB_ValS_R15Z侵染不会导致宿主菌毒力增强、不会造成养殖环境与周围水体环境的抗药性污染。
本发明的第二个目的是提供一种噬菌体组合物，其包含有溶藻弧菌噬菌体 vB_ValS_R15Z。
本发明的第三个目的是提供上述的溶藻弧菌噬菌体vB_ValS_R15Z或噬菌体组合物在杀灭和/或预防溶藻弧菌中的应用。
本发明的第四个目的是提供上述的溶藻弧菌噬菌体vB_ValS_R15Z或噬菌体组合物在制备抗溶藻弧菌的药物中的应用。
本发明的第五个目的是提供上述的溶藻弧菌噬菌体vB_ValS_R15Z或噬菌体组合物在制备抗溶藻弧菌的水产饲料或饲料添加剂中的应用。
进一步地，所述的溶藻弧菌为Vibrio alginolyticus ATCC 17749T。
本发明的第六个目的是提供一种抗溶藻弧菌的药物，其含有上述的溶藻弧菌噬菌体 vB_ValS_R15Z或噬菌体组合物。
本发明的第七个目的是提供一种抗溶藻弧菌的水产饲料或饲料添加剂，其含有上述的溶藻弧菌噬菌体vB_ValS_R15Z或噬菌体组合物。
本发明的第八个目的是提供一种体外防治溶藻弧菌有害感染的方法，该方法是利用上述的溶藻弧菌噬菌体vB_ValS_R15Z或噬菌体组合物对溶藻弧菌的裂解作用在体外杀灭溶藻弧菌。进一步地，所述的溶藻弧菌为Vibrio alginolyticus ATCC 17749T。
本发明具备的有益效果：
本发明中的溶藻弧菌噬菌体vB_ValS_R15Z是一株隶属于Autographiviridae(科)、 Kaohsiungvirus(属)新的裂解性短尾噬菌体，对溶藻弧菌ATCC 17749T具有高度专一裂解作用，能够有效地杀灭、防治相关的弧菌病害，在该噬菌体中未检测到毒力因子基因和耐药性基因，该噬菌体无毒副作用、安全性高、环境友好，表明溶藻弧菌噬菌体vB_ValS_R15Z 完全可以作为水产溶藻弧菌病害杀菌剂。
本发明的溶藻弧菌噬菌体Vibrio phage vB_ValS_R15Z于2021年12月30日保藏于广东省微生物菌种保藏中心(GDMCC)，地址：广州市先烈中路100号大院59号楼5楼，邮编：510070，保藏编号为GDMCC No：62170-B1，分类命名：Vibrio phage。
附图说明
图1为溶藻弧菌噬菌体vB_ValS_R15Z的噬菌斑形态。
图2为溶藻弧菌噬菌体vB_ValS_R15Z的电镜照片。
图3为基于DNA polymeraseⅠ构建的溶藻弧菌噬菌体vB_ValS_R15Z系统发育进化树。
具体实施方式
以下实施例是对本发明的进一步说明，而不是对本发明的限制。所用的方法及技术如无特别说明均为常规的方法及技术。
实施例1：溶藻弧菌噬菌体的分离与纯化
采自福建厦门第八市场废水水样样品，经0.22μm滤膜过滤后冷藏状态下(储藏于4℃冰箱中)带回实验室。
为增加噬菌体丰度，提高噬菌体分离成功率，在进行分离实验前，对样品中可能存在的噬菌体先进行扩大培养，具体如下：向10mL对数生长初期的宿主菌(溶藻弧菌ATCC17749T，分离于导致人类食物中毒的腐败马鲛鱼，对水产动物和人类皆有致病性)培养液中加入2mL经0.22μm针筒过滤器再次过滤的水样，放入摇床继续培养24h后，回收混合培养液，经0.22μm针筒过滤器去除体系中的细胞与杂质，得到扩培后的混合噬菌体液放入4℃冰箱保存。
取1mL前述混合噬菌体液，用SM缓冲液(按1L超纯水中加入5.844g NaCl、2.4647 gMgSO4·7H2O、6.057g Tris、0.100g Gelatin的比例配制SM缓冲液，配制完毕后将其pH 调整到7.50，100℃高压灭菌30min，冷却后使用)进行梯度稀释(稀释梯度为×100、×10-2、×10-4、×10-6)，保证样品中的噬菌体数量适合。在已稀释的各梯度噬菌体液中，各加入1mL处于对数生长初期的宿主菌培养液并摇匀，常温、黑暗条件下静止15min，使噬菌体吸附感染宿主菌。
吸附结束后，向混合样品中加入5～6mL融化的2216E半固体培养基(按1L超纯水加入30g的2216E液体培养基粉末(海博生物，青岛)，煮沸1min以溶解粉末，冷却后经 0.45μmMCE水系微孔滤膜(津腾实验设备有限公司，天津)过滤，滤液即为未灭菌的液体培养基，备用，向其加入0.5％终浓度(wt/vol)琼脂粉末，121℃高压蒸汽灭菌15min，冷却备用，此即为2216E半固体培养基，使用前再次重新加热融化，使用过程中控制温度在50℃)，快速混匀后倒在准备好的单层平板2216E固体培养基上(向未灭菌的液体培养基中加入1.5％终浓度(wt/vol)琼脂粉末，121℃高压蒸汽灭菌15min，在培养基降温凝固前倒至直径9cm的培养皿(Biosharp，中国)水平放置，冷却凝固备用，即为固体培养基)。待上层平板冷却凝固后，倒置于30℃恒温培养箱培养，3～4h后即可定时观察是否出现噬菌斑。
双层平板上出现噬菌斑后，仔细观察并按形态大小分类记录噬菌斑，用灭菌的剪口枪头挑取单个噬菌斑，放于1mL的SM缓冲液中。噬菌斑于4℃避光保存过夜，使噬菌体充分溶解于SM缓冲液中。所得噬菌体浸出液进行梯度稀释(稀释度为×10-2、×10-4、×10-6、×10-8)，重复双层平板实验，纯化挑取获得的单个噬菌斑(图1)，获得单类纯化噬菌体。
实施例2：溶藻弧菌噬菌体的扩培与富集
取1mL纯噬菌体液加入到10mL对数生长初期的宿主菌(溶藻弧菌ATCC 17749T)培养液中，30℃摇床中培养。细菌培养液出现明显裂解现象后(即培养液恢复透明且有细胞裂解残留的絮状物)，回收培养液并用0.22μm针筒过滤器过滤，保留滤液。将上述滤液加入对数生长初期的50mL宿主菌(溶藻弧菌ATCC 17749T)培养液中，继续培养至出现明显裂解现象。重复过滤过程，滤液加入200mL对数生长初期的宿主菌(溶藻弧菌ATCC 17749T)培养液中继续培养。按此方法进一步将培养体系扩大至1L。
1L培养体系出现明显裂解现象时，加入DNase I和RNase A(Takara，日本)至终浓度为1g/L，混匀后室温消化30～60min，以降解体系中游离的宿主DNA和RNA。加入NaCl 至终浓度为1mol/L，充分溶解后置于4℃(或冰浴)1h，使噬菌体从细胞或碎片上脱落至液体中。离心(5000rpm,4℃,5min)，取上清液经0.22μm过滤，除去体系中的细胞和部分碎片，得到1L噬菌体液。
向上述噬菌体液中，加入10％(wt/vol)的PEG8000(生工生物，上海)，混匀使PEG8000 完全溶解，4℃下避光处理1～3d。使PEG与生物大分子充分结合聚集，形成聚合物沉降。沉淀完成的样品离心(12000×g,4℃,1h)后弃上清，残余的沉淀物即为噬菌体沉淀。向其中加入SM缓冲液重悬沉淀，使噬菌体从PEG沉淀中重新溶于SM缓冲液，转移至15mL 离心管。4℃避光放置12h以上，使噬菌体充分重悬进于SM缓冲液。若重悬后的样品杂质较多，可加入等体积氯仿(用以除去脂类、细胞碎片以及其他杂质)，轻微摇晃使两种液体充分接触混匀后，低速离心(5000×g，4℃，5min)分相，取上清，得到噬菌体浓缩液。
将CsCl粉末溶解至SM缓冲液中，调整溶液密度为1.65g/mL、1.5g/mL、1.4g/mL、1.3g/mL梯度溶液。向离心管中缓慢地依次加入密度梯度溶液6mL，在密度梯度溶液之上加入全部噬菌体浓缩液，用SM缓冲液使离心管注满并配重。在200000×g、4℃条件下进行 24h的密度梯度离心。离心结束后，可观察到蓝色或白色的噬菌体条带即为噬菌体纯颗粒，小心取出条带于离心管中。取出的噬菌体条带至30kDa超滤管中，离心(5000×g,4℃,5 min)透析，并加入SM缓冲液冲洗，以去除CsCl。完成后将噬菌体溶液转移至1.5mL离心管中，获得高浓度富集的噬菌体溶液样品，于4℃避光保存。
实施例3：溶藻弧菌噬菌体的形态观察与鉴定
取20μL浓缩后的噬菌体滴至200目覆有碳膜的铜网上，黑暗吸附10～30min后，用1％的磷钨酸负染色约20min，干燥30min以上，使用JEM 2100型透射电镜，在120kV电压下对样品进行观察，图像使用GATAN INC CCD图像传输系统进行采集。
如图2所示，溶藻弧菌噬菌体头部长度为69nm、宽度为55nm，尾部长16nm，根据国际病毒分类委员会(ICTV)最新病毒分类系统的标准，该噬菌体属于短尾形噬菌体。
实施例4：溶藻弧菌噬菌体的基因组分析
首先提取DNA，采用酚-氯仿抽提法，具体步骤如下：
向溶藻弧菌噬菌体vB_ValS_R15Z悬液中，分别添加1.5μL的100mg/mL蛋白酶K溶液，100μL的10％(wt/vol)SDS溶液，10μL(0.5mol/L,pH＝8.0)的EDTA溶液，混匀后置于55℃恒温水浴中进行消化3h。
向消化后的噬菌体悬液中，添加等体积的苯酚-氯仿-异戊醇(25:24:l，体积比)混合液，混匀后离心(12000×g,4℃,5min)，把上层水相转移到新的离心管中；这一步操作需重复两次。
向收集的上层水相溶液中，添加等体积的氯仿-异戊醇(24:1，体积比)混合液，充分震荡30s，离心(12000×g,4℃,5min)收集上层水相溶液。与等体积异丙醇液体进行混合，混匀，-20℃静置l h，再次进行离心(12000×g,4℃,5min)，收集离心下来的噬菌体核酸沉淀，用事先预冷的70％乙醇溶液冲洗两次，风干。
将该核酸沉淀溶解于100μL的TE缓冲液中(10mmol/L Tris-HCI,1mmol/LEDTA, pH＝8.0)，置于-20℃保存，备用。
噬菌体全vB_ValS_R15Z基因组使用Illumina HiSeq4000平台测序，原始数据用Trimmomatic v0.32整理，并用Velvet v1.2.03软件组装噬菌体基因组。基因分析显示，该溶藻弧菌噬菌体基因组长度为43873bp(其核苷酸序列如SEQ ID NO.1所示)。
基于DNA polymeraseⅠ构建系统发育树可知该溶藻弧菌噬菌体与Vibriovulnificus phage SSP002分类地位相近，隶属于Caudovirales(目)、Autographiviridae(科)、Colwellvirinae (亚科)、Kaohsiungvirus(属)(图3)。结合电镜观测结果，根据国际病毒分类委员会(ICTV) 最新病毒分类系统的标准，将其命名为溶藻弧菌噬菌体Vibriophage vB_ValS_R15Z(于2021 年12月30日保藏于广东省微生物菌种保藏中心，保藏编号为GDMCC No：62170-B1，保藏地址为：广州市先烈中路100号大院59号楼5楼，邮编：510070，分类命名：Vibrio phage)。
在核酸序列水平上，将Kaohsiungvirus属的噬菌体进行全基因组比对，结果显示vB_ValS_R15Z与目前公开的噬菌体相似度很低，在86.6-90.2％之间(见表1)，表明溶藻弧菌噬菌体vB_ValS_R15Z为一种新病毒。
表1 vB_ValS_R15Z噬菌体全基因组在Kaohsiungvirus属内相似度比对结果
此外，在致病菌毒力因子数据库(VFDB)中进行噬菌体的毒力基因检测，和在综合耐药性数据库(CARD)中进行抗药性基因检测，未检测到毒力因子基因和耐药性基因，表明该噬菌体不会增强宿主毒效强度、不会造成周围水体环境的抗药性污染，安全性高、环境友好。
实施例5：溶藻弧菌噬菌体vB_ValS_R15Z的氯仿敏感性检测
取0μL、20μL、200μL氯仿分别与纯化后不同丰度的噬菌体vB_ValS_R15Z溶液混合，调整噬菌体丰度为103、105、107、109PFU/mL，混合后振荡30s，然后于25℃静置30min。经低速离心(5000×g 45min)后分相后，小心取出10μL表层上清液，滴于事先准备好的宿主菌(溶藻弧菌ATCC 17749T)双层平板上，于30℃培养箱中静置培养，实验设置三个平行样，同时用SM缓冲液代替噬菌体液作为阴性对照，定时观察是否有噬菌斑出现。溶藻弧菌噬菌体 vB_ValS_R15Z的氯仿敏感性检测结果见表2，氯仿处理后的噬菌体依然出现明显的噬菌斑，表明噬菌体对氯仿不敏感，噬菌体无脂质结构。
表2溶藻弧菌噬菌体vB_ValS_R15Z对氯仿敏感性检测结果
注：“+”表示能够侵染出现噬菌斑，“-”表示不能侵染未出现噬菌斑。
实施例6：溶藻弧菌噬菌体vB_ValS_R15Z的宿主范围检测
选用了弧菌属多个常见种的海洋浮游细菌和水产动物病原菌(表3)，包括V.alginolyticus(3株)、V.parahemolyticus(7株)、V.owensii(4株)、V.campbellii(2株)、V.harveyi(2株)、V.chagasii、V.cholerae、V.fortis、V.inhibens、V.variabilis。用2216E液体培养基将待测弧菌培养至对数生长初期，与融化的半固体培养基混合制备双层平板；将 10μL无菌的高浓度(108PFU/mL)溶藻弧菌噬菌体vB_ValS_R15Z液滴于凝固的双层平板上，于30℃培养箱中静置培养；定时观察是否有噬菌斑出现。实验设置三个平行样，同时以溶藻弧菌ATCC 17749T代替被试菌株作为阳性对照，用SM缓冲液代替噬菌体液作为阴性对照。
如表3所示，溶藻弧菌噬菌体vB_ValS_R15Z对Vibrio alginolyticus ATCC17749T具有高度专一性。
表3溶藻弧菌噬菌体vB_ValS_R15Z宿主范围结果
注：“+”表示能够侵染，“-”表示不能侵染。
以上仅是本发明的优选实施方式，应当指出的是，上述优选实施方式不应视为对本发明的限制，本发明的保护范围应当以权利要求所限定的范围为准。对于本技术领域的普通技术人员来说，在不脱离本发明的精神和范围内，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 中国科学院南海海洋研究所
南方海洋科学与工程广东省实验室（广州）
<120> 溶藻弧菌噬菌体vB_ValS_R15Z及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 43873
<212> DNA
<213> 溶藻弧菌噬菌体vB_ValS_R15Z(Vibrio alginolyticus bacteriophage vB_ValS_R15Z)
<400> 1
gccgcctgcc cttcccacag cctcagagag cggctgtctc acagcgtgag actacttgac 60
cgcctgaccg ctctcgcccg tcctgagcgg acacagcgcg tcctatccct atagtgttat 120
aacataacat tacgcctcag ccgctctagt gttataacat aacaattcca agtacgaacc 180
aagcctatcc aatagcgcac caagagcgag aaagagcggc gctagccatt atccatgcaa 240
tcgctcaggg ataaggccaa gtccgctctg agacgctttt ggagccccta tggggggact 300
gtcgattgcg cgagcgggag gaaaccgctc acatgtacag ccaaaaatat tttagccccc 360
tatcaagtgc agcaggtaag ctccaaggaa gccacctacg aatactacaa taacttcttc 420
tagtgtactc attctccaac ctctctcttg taccacagct tagcaagctc caagtacgtt 480
aaggaactct ctttgtcacc agccaaggta cgctctaggg ctttatttag gcaccagtca 540
gtacaggtca tttgtcattt acctccgtat caactgcata aaccttacaa gcccacttac 600
ttaggtactt gtgcattcgc ttgattaaat ctagatggaa cttagatgca tctttgtccg 660
gtacatccaa gaccaaggtt actttaatac gcttcttctt aatactcatg gtcttcctcc 720
attatcatag aactgatgat taccgcaagc atatgtctta tgcatcttgc gagtccagta 780
gttgtctaca tccttacgag catagtgcag ttcagtgcta gttatcgtat tcccacttaa 840
gtaattgcta tagccttcaa gagcagctag tgcgtcagtg aaggtgataa ttgagtggtc 900
agcaggatta tctttaccta ttgtccaaga gaactgctta ggctgcaaga caacatcttt 960
tacagtagaa gggtagaaag aagaatctac ccttcgcata gttacttcag ctactgctac 1020
ttggcactcg aaaggctcac cacgagcctc atgatgcacg ttaagagata gccatagaat 1080
tgcttcaacg aacatggcta cctcctattg cttagcctct tcgtaattta atgaagtcag 1140
caactaaacc ccaaggggta cggacacacc accaagcaag tccatatagt aggatgtact 1200
tgataggaat gtttacgtcc atgttgatga cttccccaat gctctgtgac tggatagcct 1260
gtgctgtgtc agttgcagct tgagctactt ctcctgtagc ttctggcatt gcaactagag 1320
agctacaggc aacagtcgag aaagcagcca gagcgactgc tatgatactc ttattcatac 1380
aacattaacc cacaactcgg atagtgcctg agtcgttcca aatagcacca cttggcaaac 1440
cagtagggga tgttggtaac actattgtca tagtccctcc cttaataact gataccaatt 1500
ctgtgtccct cccatatatt tcagtctgat aagtactacc tacagaagtt aaagatgagg 1560
tcttcacaga gaatacttta ccagcaccaa gacccgatga atcactctgt tcatgataga 1620
actccccaac agactgacct gcacctacag atgcatcttg actcaaaata cctatcgagc 1680
caccctcatc ctttatcacc ctcactgtcc cattgtattg acccagctta ccacgagcaa 1740
atagagtatc tgctgagttc tcctgtgtca caaaatcacg agtaatacct ttgaactcat 1800
aaccatctac tgtgatatta ggtgagccgc cagcctctgg agcgatatag ataccataca 1860
gcaatggatt acttcttgtg tctatgtaga ggtcagttat gctaatatca tcacaatctt 1920
ctgcaatcct gcatacatcg ttaccaataa aggtaggtgc atctagtatt ttaagattgc 1980
tgattgaaat gttcttattc ctagccaagt aaagcgggcg ctctgttgta caacctttta 2040
tggttatatt agttgcctgt aggttcttaa caccagaata ctcagtgtta gttgattgcg 2100
ccataccaga atagcatttg tagacagaaa tattactgag ttgataaccc tcacctgagc 2160
ggagagccaa tgccctagac gtagattccg atattgtacc gtcgtttata actatacctc 2220
tgtccttacc atccgcattt agaactatac cctcctgagt agcttttaac agaaagccat 2280
tgaatgtaat attgcttaca tcttctatca ggtaaagtcc gttgttcctt gtgttcacgg 2340
cttcaacttc gctgaaatgt ataaagctag attcaccacc tacgtgatat gccttatcac 2400
tttgtgttac gtatgaatcc ttaacccaac aatgatgtga gtctgtaaaa gagagagact 2460
ggtactctga actagatata tcatcatgct gataaatgcc gtaagcccta aatttgttta 2520
tatgaacaaa ggagcttttt agtacaagga acccgcctct gtagttttct tccaattgct 2580
tcagagggtt tcttactttt atatcttcga aagagcacga gttcatccac gacataaatg 2640
cacatgattg gaatttaaag ttagcatcaa gcgtaactcc atgaatacct gcatctgtta 2700
ctaggttaga gcttgttcca cttgtgccta tcatggtcgt caggtcgcca ctcactttgt 2760
ctgctgctaa gatgaaagtg ctattgtgac cctctcctat aaccttagtt ccatttgaga 2820
ggttaatcct tcgtgatacc ctataagtac cagaatgaag ctttacacga cctccatagc 2880
tgaacaataa ttctagtgct ctggtgtcat cagcgatacc atcaccaact gcaccaaaat 2940
ctttaccaga tactccattg tctagaatct tatcataaag gctcctcaaa actgcgtcag 3000
caacactgag cttgaaggta gcccaccgag cacgaggctt ggttaattta gtgagtgcca 3060
agttgcacct ccttattaat agtattagtt acattcataa agccctctct aagaaagagc 3120
ttgagtatgg aactggtacg agcctccggc ttggaggtac acgccacatt cattagtgtt 3180
acttcatgat taccttgtac gacattgccg atgcaaggtt aggttacgat acacctcatt 3240
atactggtgt gatatcagta gctccttttg ttactgcaat ccatttgtca gtcggagctg 3300
aacctgccgc ttgtagaagc acaggagcgc ctgatttaat gatggtgatt agggagccaa 3360
ctcgcttacc tgattgtgtt gcatcattca cgatatgttc tttgtctgca aggtgagcct 3420
ctgccaccac tggcatgtta tgaagtgtta caccttcttt ctctactgct gtgaagttct 3480
ttgcgtcacc tgtaattgaa cctactactg ccatttgtat atctcctatt gattagtata 3540
tagtatagag aatagaagaa gaaaggaggg actcttagga gtccctctag ctgagtgctc 3600
tagatgctct agatgctcta gtacgctctt tccctcttat acgctctcct atagggcaac 3660
ttaattctaa gtgcctgatt tataaagaaa tttagctatt gctaatttac cattttcttc 3720
gtccagctcg gcgtttatta ccgaatctat tgcctattct ttgtacagcc ctagctgtcc 3780
ctagagcggc gtctccacgc attccgagtg taaggttagt catgtattgc ttcatcgccg 3840
tcgggtcgct cataacgtca taccagcgtt tctgctgctt gatttgacgc tgcatcacga 3900
ctctcagttg gtcatagtcc agattctcaa caatctgacg aactgcacca gccagagcgt 3960
caaggcggtc atcatgtcgt agacagtctt tgtcacgcgt gatatttgac atctggtgga 4020
acagagagaa gttcattcgc tgttccgcac tatatgcttg gattagctcc atatccttct 4080
gtacgacctc tgggcgaacg attaggcggt gtgtggacat cactggctca agaacgtcga 4140
taattctcag ctctttctgc ccagactctt gaactgcctc aagcgttaca gggtaatgtt 4200
tctcgaaata aggcttgata gcagcaacgt gcgcaccatt accatagtta ttctctatga 4260
atacggtctt gcactctgca ttcttcgctg ccattacaag atacatgagg ctttcttccg 4320
aatagccacc attagttgca ccaatatcat agatatagat agttgttccc atgagcttga 4380
tgatagcgta agccgtttcg tccccgttct taccgccacc agcagggtca atgtacatga 4440
tggtacgctc aaagtcgccc atctcgtagt caacctgcat agggtagtag aagttatctg 4500
tcttacgagt accaaactta ggcgctcctt cccagatgca ctgtggtgag ttacaccaga 4560
taggcataac ctgagcacgt tctctattga aaggtgcaac tatcaagtca ctaagacgta 4620
atgggtaacg gtctgcgtca gtcaagccag tgttaagcat gaactgtagc atgaacttag 4680
ccttaccttg agcaagctcc ttctcgttaa gaaggtcatt atcatacatc tcagggcaag 4740
taggttgtcc ccatgtacca tcaagaccac cgccagtctg caacgtaggg tcttctttga 4800
tgtccttaag aataatagga gcaagcttgt ctccatagta ttccatctgg ttctggttag 4860
ggtaacgccc tgtccatata cgaaccttat agccacgact tggtagccag ttatagattg 4920
actctactga ttgcggtgta ccaagataga tgatgtcacc aaattggttg attgattcaa 4980
actcaagagt ctgctccatc agccattcac gaccacctac agtacgagag ttctgcaatg 5040
actcgatatc atctgcaatg ataatatcag cacgagcacc ctgcgcacct gactcaatag 5100
agtagcaggc aacagaaggt gatttatcac taccgcgtaa gcaccagtgt atatcgaatg 5160
actcgataga gccacggtct ccagcgtact tgtcaggacg catgaactca aggaaatcta 5220
gtgctgagaa tatcttgata atccaacctg ctatttcctt agctcgctta gagttttgag 5280
agaatacaac tatacggtag tgcggattat gtatgagcat gaacaccgca tagatagcag 5340
taagcgtggt cttagcctga ccacgctgtg ccataatcat acgatactta tgccctacta 5400
gtaggaactt tagaatgtct gcctgtatcc tgttaaggtc tgggttccct acgatagttg 5460
tgttaataac aacttgagca aagaagcaca aaccttcaac ggtattagga aacatctctt 5520
gaagctcata cagcatcttc cagcgttcta cctgttgctc tgctgtaaga tttctagcca 5580
ttattcatcc tcatccaaga atggaataac gttatcatct gtaccagttg cgcgagcacc 5640
cttgttcttc tgagcttcct taatctgctt gagtcgctta gatagctcag tctcttcatt 5700
catctcagga gctgcacaag taatgctgtt ctggtctgct gcccatttac ctgctgctgc 5760
aagtgcctta ccatctccaa taacatcttc aatgttagca ccttgattta caagtttgag 5820
catctcttct agccgctttg agtgcaaggt agtaactagt ttgtgtacta gccctacctc 5880
atcctcggta gcagcattct tattcggtct gcggtctgct gccatttaaa tcccctttct 5940
ccacttgtat atgatgtatc caatctggat agccagatag aacagagttg ctattgatac 6000
aatgtcaggt aacgtgaagc cgtagaactt cataccaacg taagccgcag gtggtgcggc 6060
tggtagagtc tcacggacaa gctccgttgc cttaataatc ttttcactca caatacctcc 6120
ctaagcacta tgcttctcag cgtaccacat atccacaacg tcaccttctt caagtacctc 6180
agctagctgt acattatcac caaccatgac atagctatgt gaaggggctt gatatacgcc 6240
atttacgaat aaagcagtaa agtcatgctc aggcgctagg aatgagtctt caccacctgt 6300
agctgtgtac ctataatcgc tgaatgtaga acctactgca agtttctcaa gagagcctag 6360
ctttgcgaat acttcgtcac cttcttcaag aggctctgca aagtaaatta ttccattcat 6420
ataattgaaa gacttggttg ggatttgttc aactccgtta atagaaagct cgcaatagtt 6480
aaagttatat ccagtatcca ctgacaactc tccgccgacg gctgtacgtg tgaacggcac 6540
taagtagttg accgaagaca caggagtgga tgattcaaga ttagataatc tattatctaa 6600
ggaatctgtt acagatttct taatagcatc atctgggtca actgcatccc ctaagttctt 6660
aggcacgttt ccgccaaaat ctatatcttg cttcatgtag aagccttctg gtacgaaacc 6720
atcaataagc tcttgtgtaa gatacaactg ctgcaagaat gaattgttaa cttgcctgtg 6780
cccaaagtta ttaccacgag caaagtctgc atacggagca tccacaggca tctcccttcg 6840
tatcagtaca tccgaaccat cagtaggtgc aggcgtaatg attactaaat gtggagattc 6900
tggttgaata acattagcta cctcaacacc gtcaacatat gtgtgtatgt cttcggtgcg 6960
gaagtaaccc atgttttcac ctgcgacagt gagcgggaac tgtgtacgtg acccgtcccc 7020
tacatgtcgc acaaacgata ggttactatt cgccatttat ctctccttat tctgttatct 7080
atagggcaac cttattcagt tgccactcca aatgtcaagt tccatagagt agaattatac 7140
caaggcaaca ccctttgtat tgtcttaata gaacgctctt cctctcgatt tgttttatcc 7200
tcggttgcta gagacatagc cgccgatgtt gccttagcta catccttagc catttcaact 7260
gactggatac cggagaagtt accagcacct ctcatatctt ctggcaagaa gttaagagag 7320
ttcagcattt gtaatggaag cataggtgca gctaggatgc ctacacggtt aattacaccc 7380
cagaatagtc cttcatcctc taggtgctta cggatgtact catcacgttc acgtcctgag 7440
tactggttag cttggatgta ggtatttgcg acaagagcaa agtagccaag aacggcttga 7500
cctgcaccca tcgacaacag aagagcacgt tcattcttga tgccacgtaa aagcatcttc 7560
tcatacgaac caatagtaaa gctcaacaat gtggtcataa ctttaccaag ttctttgttc 7620
atgtaggcag gttgctcacc aatacccaga cgaaggaacg aactacctat cgtattacga 7680
acagcagtgg ataatgagtt gtatgtatca gcatctagct gacgaactgc attaaagata 7740
tccatatcct tgttatctct taggtgcttc attacagcat cgacctgttc gctggacata 7800
ccaccaacac gaataagtga gttacgaacg ttgtctgtaa cctccccttt gattgccata 7860
tctctcaggt tattaacgat agctctagct gtcatctcct caccaccttg ctggaacatc 7920
ttgaaaccgt tcacggtcat tgtgatatcg agagcgttac ctagcatgtt gtcaatacgc 7980
tttagcttgt tctttgtcat atcatcgaag tctgctccat tgtagaactt acgtccgaac 8040
agatactcct gatgccctgt agcagagaat gtacgggcaa agtccatcat aaacttatcc 8100
tttgttacac tacgtgttct gtaatcgaac catcgtgatg aaggaatctg cttaagcgtg 8160
tttacaattc cgtttctcag catggcaact gagtactcag gtattgtcat aagaccagta 8220
ctgcgtagac gaactactga cgtagcctta cgagccatac gagtaacgtc cttcatacca 8280
tcatggtctt ctagtggctc tttgtaaaga agtcgaagtg cctcatccag cttatcagac 8340
catcttccgt tctctaaatc ggatagtacc tcttgagcct tagctttagc cttgccatcg 8400
agacgggcta catcaagacg aatgtcgttt atagctgcat cacgcgctgc tgcaacagct 8460
ctttggaact gatgacgaga gtggaatcca ttactagcaa gacctgcgtt tgcagcagag 8520
tcagatacgt acttcatgtt tctgtcaatg ctcgtatcta tcaggtcaac catccataca 8580
tcaccagagc gagcttttag gtttggacgc aatgagaaca tagcacgagg tgagatttgt 8640
cccatctctt ctttatcaaa catagcttgc ttaatatcgt caataacatc tttgtcaatg 8700
ttgttagcta gaagttcttc ttcaagcttt ctgaactcag catctgacat aaagctatcg 8760
aaagtcttgt gagatttacc accatatgac atcgcacgag ctacttggtt cttagctagt 8820
ctaagagcat tctcacgaga cagtttaata ccacctgtct gataagcaga tgcaatacag 8880
tcatagatga aatcagcgtg gttctcatta gctatgatat tagtaggatt aaacacaact 8940
gagtgatact cgtgtcggtg cttaatctta tcgaaaccaa ccacgttgta atccttgttg 9000
ttcttaagac cttgctcata caaacgagaa cgagctttag cgccaagaat gattgcgtca 9060
tcaccttcta ctggattatt gctaagcaaa gtaccagtag cttgcttcat aacagtctgt 9120
ctatcaaact ccatctgtag ttcattaagc ttaccaatat cgaacttgcc aacacctcgc 9180
tctttacgga aagcatcaaa tgcattctga taatcaagat aatcaggtac agctctgttg 9240
aacaaagtct ctgcaagctc ttccgcagac tgatgacctt cgatagtacg ttgtgggtct 9300
ttcactaaga acttaccaag accacgagtt gcatcatcag gtgctgtgtc aatctcagag 9360
gacaatgagc gtaatggttt accaatagct gaagtctcga aacgtgtgtt acgaggtact 9420
ttagcagcaa atgcctctgc atcactaaga gcgttctgaa tctcttcaac atctgtcggg 9480
gtgagaagct catcatatgt ctgtatatca tcaaattctt gtcttgctct cattgcacca 9540
acgctttgag aaggtggctc ttccttgccc gtctcatcta ctactggctt acgaccagcc 9600
ttgatgtctt taccaagaga ttttgaaggc tctacgtcaa attcaccacg ttctgcctta 9660
agttctctgt atcgctgcat gagtgattca ggaaccttgc cctgctgaag agcatcaatc 9720
tttgaattag tagccatagc aattctgttt gcttcaattg catcattaag ctcagcttct 9780
cttctagcga ttgcaccttc tacttgctga atctcagcag ctcttgctga aggtcgaagg 9840
ttagggcgag ccttaatctt ctcaattatt tcgagacggc tcttacgaaa ctctctgaac 9900
tcatcacgaa gtttcttaac cttcttctgt gacaggacat caacacgctc accagcttcc 9960
ttacgaagtt catcaattac atctcgctca gttaggtatc gtttacgagc aacagggtct 10020
ggttgctgtg atgcaagagc ctcatctgcc atatgataac ctttgacatt ctcctgtgtc 10080
ttaatctctg ctctctgtat actctcaaca gcattagttc tgtcaagtgc attacgagct 10140
gccttgtaac ctagcttagt agcatcaatg gcacctgaga aagctacgcc gccaatggct 10200
gacatgtaaa ggtcttcctc agtcctctga gtgtcaccca tcttgagcaa gtactctgat 10260
aatacaccct cagagccacc acgtaacata gaccagccaa ttgagcctat acgacctacc 10320
ttattagtaa gagcagcagg tgcgtcaaca aacatcaggg gaaccatagc agggtcgaag 10380
atagcagatg ccgtttcaat agcatatcca gtaactccgc cattctcaag aacctgcttt 10440
acctctctgt ctgctttgat tctgctttgt ctatgcgcaa agttctcagg tgaaacagca 10500
ccagccaaga actctatctc ttgctcggtg taaccttctt tgttgtactt cataagttca 10560
gctctgtcta ccttgtagtc ttgctgaaca ccgccctctt tctctgcttc ataatgtcga 10620
gacagagact cacctaccca atttcgatac ttagctgcct cccacatttc agagagagag 10680
ggctgttcgc cctctggctc tggtggttgg taatcttcgc tcagtataga gagtgggata 10740
tgagatgggg tataatcaag aaactctctt ctttctggca ttatctatta cctctttgtt 10800
tgctccaata ttcatagaac agcgccatat gtgctgcgtc ctcttctcgt tgacgctctt 10860
cttctatacg ctgttcacgc tctttctctg ctgttgtagc tatttcttca atacgctctg 10920
cttcaccttt ctcaccataa cgcttaacat cactcataag gaaacgagca tcgccgactt 10980
gttcaccaaa cttatcaacc ataatgaatg tagtaccagt atcatttaca acaaactcta 11040
tatcttctgg ctctacgcct aacgggttga tatcctctga tgacaggacg tttgatgcaa 11100
agtgtttcat gtatctgtca acaagctctt gtcttactcc aatagattgt gcaagctcag 11160
gcttggtctt gttaatcagg gtattgttga atgtctgaga ataattagct agctttgagc 11220
ttgctgctgt tttagcgttt tgttcaacat ctaaaccacc tcgatacaag aaggactgtg 11280
cttcttcacc tatgatatta gacagcttat cacgttgcca cgcaggtaca tccttcttgc 11340
cgaactgata ccacttagca tttagctcat cttcgataac atctgaaact gcatcacgtt 11400
gctcagtacg ttgttcattg ctgatgttgt acggattacg tctgatgtca taagccctac 11460
ggaaagatga ataaggtgag cggttcttca tacctgactt gatattcata gccatagcca 11520
agctatcttt gtcagtgaag tacatagaca acgaagtgtc atccatcatt tggattatct 11580
taagtgcatt agtagcatac gtaggtagct cttcatccgt agtctcttct gggtttaggt 11640
tgataagggc ttctaggttg tctttgagac taggtatagg cattcgctga actctagacc 11700
agtcaagctt ttctttaaga attgctgtat tcgcatcagc ttcactatga ccttgttcga 11760
ctaatgactt agccttttca gcccaagcac cctctagttg tttaacataa tccttcttct 11820
ctttcggtgt gtacttaccg aggagaccaa gaggtatagc attctcatca gtgaaagcct 11880
ttgatgagtc aatagctata ttagcagttg tcttttgctt agctattgca gttgctctct 11940
gcttcttaag agacgctatc cgttccttag agtatgtgtt agggtattgg ttgttaagtt 12000
tttcaatctt tcgtaacgta gactcccaag aagtactacc gctcagtgcc tgtgtctcta 12060
tatccgccat tgtatcacca atgacaatag cattttgctt agctacgaaa gagtcataat 12120
ccctctgcgc tttatcaagt gcaggtaggt tcttagacca atcccttgcc tgcaactcag 12180
caagcagtcg gttgtcacca tcacctgctc tagctgctgc ttgattaacc agagcagact 12240
ttagttcaaa ctcattctga ccaagtgctc ttgcacgagc ttgtatctct acatctacta 12300
agtgtttaac atctgcgtta gggtttgcaa gtaagtcgtt aatactaata ctaacagctt 12360
cactcgcctt cttagattga tactctttct taatacgctc ttcagcttgt acaatagaga 12420
actgagcgtc ttgtacttga agtgaaacag ctttcaaagt ctgttggtct gttccatact 12480
tatctaagat aggctggaat atctgggttt tcatttcact gatagtctgc tcatcggcat 12540
ctggattttg gttataccac tccaagaact gattaccagc ttccatagta tcgtgcttac 12600
caacaatcat gttataggcc aactgacctg ccttagttgc gtcttcagtt ataccctgac 12660
gctcttctgc tgtcttcatc aagtcttgag cagctcggtt agcttgctta actttgtcag 12720
cttctatctt cttctgaact tcagtgctag ctgctgcgta tccggcttct gcaaacttac 12780
caagagcacc taatatctgt gagcgagtat ctacagttct agtaggtgtt gcaacctctc 12840
tggtttgttg ttggaaccct cttcgggtcg ttcttttcat ctcttgcaat ggcatcttat 12900
actcctgact taagtgctgg ttctgctggc gctacttctg acttagcctg ttgataccca 12960
gaatagcctt gtaatgcagc agagccaata tttaataatc cagccacagc agatggtctg 13020
cttataggtc ttacatccat actacccata gccccatatc gcatagactc agcttggtct 13080
gctatattgt catactctgc ttgttggttc tgttggataa tattgaagtt actaaactta 13140
gttctctcca agtcctgtaa ttgactggca actgacatcc caccaacacc ggaataagct 13200
gccattacat taatcctacc cttctctttt aggtagtctt tctgcattga tagtgagtcc 13260
tctaatgaag cctgttgtgc ttctagctct tgctcagata actcatcata gttagcaata 13320
gttgtctcaa ctaattgctg attacggcgc tctgccgcct cttcttgagc ctttgcattt 13380
tgattagctg catacataga gtaagcagca gatgcaatac taacggcagc agtaactgcc 13440
gccgccgtag tagctgctgt agaagaagca gcagccgcac cacctgctgc tgctactgcc 13500
atagtacctc cttagatacg tctgcctctt tgtttgaaca tcccttccca ctccatatca 13560
cgaagtatga aaggtctata atcatctgta acaatttcga actgtgcctg ctgagcttgc 13620
agcctaacag ggaagctgaa ctgaccttca gccaatggag agaatcctac cctattattc 13680
cagctaccaa gtcgccttcc gttgtattca tatttccatt gacggctctt tgagtacaaa 13740
tcataaacgt acacagttat gttacctatg ctttcgtagt tgagtgtaac tttgcctagt 13800
gtgaacctgt ccaaccccat aacacgacct gattggtctt taatgctagg ttgcgtaggg 13860
acaaatctag aagtgtactt agtacctgct gtgagattac aggatgtcac tccactagtt 13920
acatcagcta ggtcatcata agaccagtaa gtattaccat cagtctcgaa tatcactgta 13980
gtacctttgt cagaatccca acagccagta cctcttacaa actctagcat gtctgtgtct 14040
tcgacttcat atgggttatc ccactcccaa cgagtaccag tccagttcgc gttaaccact 14100
gcgcgttggt ctaagctaac agggaacgta agtccatcgt cgtctgcgtc attacttata 14160
ggcatttgct ctaagtaaac tccgttacct ctatctatta caaagtaaac tttgtcactt 14220
acgaacttgg cgaacataac atcaccgtca aatatccact tgtggaatgc agcctgtacc 14280
ctctgctcac cagaccatag ccagttgtaa gcgtagatgg tgtggttatt tatatctgtt 14340
cgaactagta gaaggttgat attaggactg gcgattaagt cgataggtac accttctatg 14400
tattctgcca catgttccgt aataggtctt gccctctttg tgtcagtaag agagtcagtg 14460
aacatctccc taagacctgc atacttacct gctataaacg agaacatgat agactcacct 14520
gtaacggctg gctgtgcatc tgtgttcatc gggtaggaag taacctgtct aaacactagg 14580
ctatcttttg ttataggctt tgagccatca ataaggaatt gaccgttctc cgcaaagaac 14640
acaatatcac cttcgagtat agcgtggtgc aggagattgt taacttggtc tgcatctgcg 14700
aaaacatcta tagggtcatc agatgactcc tcctgagtag tttctctgaa gaagttaaag 14760
aactcattcg tctttccaaa gatagctgac tcacctgatg tgaatagcat acggttctgg 14820
aaggttccta tactctgaat cgtgccacct acaaaagaag ggaaggggtt cgttgtgtcg 14880
tcccctacct tgcgctcttc ccactcaccg taactcagct cgaaagtacc atcagctagg 14940
cttacgagtt gatgcggcat tgtatcaagg ttgaacttat atagagtatc ctgagcagtt 15000
gactccaccc accttacgga agaacctgat tggtctgtgt cgtcgttagg tactgccttc 15060
agccaatagg agtttgcctc aaatccttca ttgttctgta ccttaactat gtagtcctct 15120
ggtgcatagg gtggcaaatt agatagctgc ctaaccttgt cctgtatagc aataaggtca 15180
tcaccgttcg cactatctac agtagatata gagaaagatg ttccatctat acgctctata 15240
aacaatgtat taccatgtac ttcagatgtg tactgactgt caggcgcacc tactgcctcg 15300
tatacaactt ctattgtatt gccttggtta ttgcctgagg atagtatgat atctgaacca 15360
cttacgctta ttgcagttgt attagagcct gttgatttgt tgtacaagct aacaacaaca 15420
cttacactac ttgataccgt aacgtagtac tcttgtgtag taccgctccc gccagtcact 15480
gtccgtctct taacactaac agtttctgta gtcttctctt tgccattaat ttggttactt 15540
agttgctctg ttatatagtt agtacgtact ttctcaatat ggcttgctac tgcaccatca 15600
ggtgttgtaa ggctggctac tttgttaccg tctatgatta tctggtaatc tctaccatag 15660
gttgcatact gacaatacac tatcgccata gaagggttag taggggtctt atcaggcaaa 15720
gtctctgtaa ccacattagt attggcaatg aatgtatagt cagctattgt cttcatcctt 15780
agagatttct taggattaga tacctttaag taatcatcag agccactatt aacattgaca 15840
actttctcat taccttctaa atcaaataca cgaggatatt cattaggctc tactagaata 15900
agataacgct ctgaaccccc tctgtcatac tgataccaaa gagagtcagg gtcagaagtt 15960
tctagtatct tcttaacgtg cctagtacct actctcttca ttagaccgta caatggagat 16020
ggtgtaaggt tttcctgctt agtgcattga ccgtctattc tgtctttatc agcttgctgg 16080
ctgacaccct gtataggtct ttcccagctt cctgttattc ttcccatgtc tacctcacgt 16140
tgttatagcc acctgctctg tagatagttt gacgagcatg agggctatca aacatgttag 16200
agcgtctctg gcgagcatcc tctgattgta gggatatcat tgaatcctga gcagcttgct 16260
tcaagttgct cattttgtta gggtcgcctt ctttgttatt ttggaaccag aaagatgcag 16320
tatctgtaat agcttgctta gcagttgttg gtaaggtatc aaacggcaga tttacaacaa 16380
gctcacaatg gagttttcca tcagctcctg tcaaccctct catatcatac ccataactct 16440
tggcatcaaa tagtctacta cctcgaaggt taacaggtag agcctttcct ctttgctctt 16500
tcaccaagca aaccaaagtg ttctgtggaa ctacaacgta accgttaaca gggtcagggt 16560
ctaacttgtg gaactcttct ttattgaacc accatccctt acctcgattg tgttgtataa 16620
cttgagagta tctatcaatc atcttatctg catcagcagc atctatgttc cagtctatct 16680
cttcctcgga gttaatacca acagagccaa tagcgtctag tactgcattt atggcatcta 16740
ttttagtatc aataccgaac tctagattga tatccatatt actccttagt ctcacactgt 16800
gagacaaaaa taagccccta ccaaagtagg caggggcaag ttattaatta agggttagca 16860
gctagtaccg cctgtacagc cgcaacgaat gcagatgcat cacctgtcaa gtctgctgac 16920
tgagttactt gtgctttacg agcggcacgt ttatcaagac cagcctcatc atcagcagct 16980
ttgattagca aacctaagtg ctcccaacgg tctgggatag cgccttctga catgtagctg 17040
tcgatgtacc aagtcttgtt acctttgttc caccagattt cgccagtaag tgcgatagag 17100
cgacctacaa ggagtgcttc actaccgaag attacagcag cagctttctc ttggtcttca 17160
ccatccgtac cagttaatac gtcatagcgg tagccgttgt tcgcattaga caggatatgg 17220
tggtctgcga cattaccatc taagtcgatg tggtcacggc tcttgttagg gaactggttg 17280
gtaggaacca caggtacgtt gtaagactta agaacgaaac ctgttacagt ctggccttgc 17340
gcagtattgt atcgagcatt gcagattcgt tctgcgtcac gcattgcgtt gaactcaggc 17400
caaggcatga agattgtaag tttacgaaca tccacgttct gtagtactag gtcttcgata 17460
acccattcaa tacctgcgat tagcttgtta gggctgaata gtgcatctgt ttctttgatt 17520
ttcttaacaa cagaaaagcc atgaccttta acacgaggtt tagtacgctt ggtcttagtg 17580
ttgttgattg caccaaagat aagttgctgt agcaacatgc ggtcttcaag gttcttaaga 17640
gcagacactt gctcttcagc tagcttacct ttaaccatga tgtcatcttg aacatcatca 17700
agaagaccaa caacgttacg ggcgataaca gttgtgtcaa tcaatagtga gtttttatcg 17760
aactccatct caccaccacg gacatcttta ccaggtgcca gtgcctgtac ttgggttaca 17820
ccaaggtatt tgttagacac gatgttagtg cctgtcactt cttgtaagtc gaagtacgac 17880
ataagaccca actgctcatg gtaagcgcgc ttaatctggc ctgtgaattt ctcaaaagcc 17940
agtgacttcg cttcaccaga ggctgatact ttagggtcta ctaaattgtt tacatcactc 18000
atgtagtgaa tctccttcta attattacgt tatctatagg gcaacttaat taaagattgc 18060
gtttaagacc agcactacga gctgcgtcta cttttgattg gtactcaaaa tctgtccagt 18120
atttatcact atccatgatg gtttggaact catcagatgt tagatatccc ttgtctagcg 18180
gagtaccagt aggtgtactg gaagtatcat caccaagtag cttgatattc ttatcaccgt 18240
tctgaagctt atcggaaagc tccatttgtg ctttcactgt ctgaatgata agaagctgct 18300
gctcatggct ctctgactcc atcactgcat tgtacgcagc aatctgcttg tcatcgaagt 18360
tcttcaagat gaagtcttcc atagcaacca gaccttcttc gccacctaca gtagtcatga 18420
actcttcagc ctgttgctta ctagttactt cctgctgttc tttgtcagca gcagcagctt 18480
taatagcttg ctcgttcatg cctttgtaca tgttcaagta accatcaacc agcgtcttgc 18540
cgaacttaga ctcaagctta gtacgtgttt cttcatctaa agagaagtca ccatctttct 18600
tgaagagctg agagagaaga gccttctcat caatctctgc ttctttcaaa gcatttctga 18660
cttcttctgg cacttctact gaaacagctt caccgttgaa gaaatactca ggtttctcgc 18720
cagactcctc tggctcttcc tgttctgtct tctcaggctc cgtggcagca gactctgcat 18780
ccgcatcctc cgcaggtgta gaggcaggag actcctcggc aggttctaca ggttccgtcg 18840
caggactctg ctctggctcc tgttctagct ttggggcttc tggctctgaa ctgccaccag 18900
ttaaatcgac aacatccgta gagttgtaat caataatttc tacatccatt atttacctct 18960
ttgtgtcagg ttgttagcta cttgcggcat tgcattagca gcgccttgca tcatcatttc 19020
ttgttgctgt gcttgcatac gttgttcttg ttgcttctgg tattgatctt ctgtcataat 19080
ccaaggagtc tcaagagaca actggctagc ggcagaacgc atgtagtcag cccatagcat 19140
acgttcctgt gctggcatag gccatgaagc aggtaagttc aacatctcag taaattgcat 19200
cagcttgtca gcatctgtca tcttacttaa tgcagagatg cctgtgataa gaacagtgtt 19260
aacaagttct ttaggtaggt cgaaaccaat acgtctaaga agcaagttga agtaaggacg 19320
ttgtagtgta ttagccagta aagagtacgc accgcctaag ttctgctcca tctcgatagc 19380
atcacggcga atctcgtaag cggtaacacg ctctgcgtta cgctgtactg ctgcacctag 19440
catgaatgct gtgcctagtc ttcgctcata cttttctagt acgtttgcaa taggggtgaa 19500
gtctgcatac ttctctaact gaagtactcc aatgtcatct atgttaccaa agataaactc 19560
accagttggc gactcaatga ggtggtcaac atccgtcaca gaccccggtt tgataaggta 19620
tttgatatct gccatgagaa tcataccctt accaagagct tctgatagga actgtaccat 19680
gtggaagtca ttagcatatt gctctaccag agagcgaccg tagtcttctc catagtttgt 19740
cttccaacgt agtacaataa acggaaagtt ctcaggtttt acacgatagg acttacctac 19800
tttctcacct acaacttctt gctcgatgtg atagaagcct tgcttaagaa ctgcacgagt 19860
gtacagctta acttcttcat catccttttc cttgtacttg tttgaggttc gcatcttagc 19920
cttaatgata gcttgcaatg ctggctcaaa tgtatcaatg cccttgcttt cttcgatgat 19980
tagttccaat agagtaccag atttatctcg tctattaaca aacctatcca atgggtagtt 20040
aacgagattc ccttcttctg gcatgtacaa gcaagtagta ccacctatca acaggtgctc 20100
cattgcctca cccaatccaa cacgtccagc tatgtgctcg tgctccataa gagcagcttg 20160
ctccgcttgg actaagccac cttgcgcttc gataaggtct tcgtcacctt ctcgtaagtg 20220
cttcttggct tcctgtgtca gctccaatct agcgaatgtg ctatgtggag ggaacagtgt 20280
catgatgagt ttgttttcga ggttgtttac acattgtgcc ccgaacgatt gccagcctgt 20340
tgtgttagtc ccagaaccct gattctcgct ggctacttca gggtaaaggt taggcaaggt 20400
ataagtactg aactcacgag cgcgagacaa gaaaggctca cgttttacct tgagcttctc 20460
atatcgtttc tgtattgtct tttcttgctt acccttgcta ttctgtttat gacgtatacg 20520
gttctttgtt aggtcaactg tatcatcttc tggtaggagt tcgatgttcc ttgccataga 20580
acctcctata cgtttagtcc agtacctgta gctgtacgtg gacggcgtaa tcgtttacgg 20640
ccttgagaga catctccggt atcttctgta cctagctgaa tgtcttctgg ctctacatct 20700
acctgacgct ctggcttctt agccggagtt ttggcttctg ggatatctgg tgagcttgcc 20760
attacttact ccttacttgt ttacgaaatt gacaacccac gtaggagtaa cctaacttct 20820
taaagaatcc ttcagtttct tcctcgtgga taccacttgc aatactaatc tgcacttcta 20880
ctgcacctcg ttcatcagcc caagcttccc aagctttcat tagcttgtac ccaacacttg 20940
tcttacgctt ctcaggtact acatatagta tcgtgtcaaa agcaagacgg gctttgctcc 21000
aaggtagagc atgacaccag ccccaaagta accccactgg ctcattgtca tcgtaagcaa 21060
gcagtaaaca accatcttcg ttctgtactg ttatcgctgc attctgcata gtgtgctcga 21120
tatcgacagg gaagttgtta tgtttagctg cctcttctgc atattgatga gctagtgggg 21180
caaaggctag taagtcaagc agagtagcct gacgtacttt aatcatggtt cgaatatatc 21240
ctctatctgt ttcttcagat tgcgcttcgt gctattggca accagcactc cataaggtgt 21300
ggtaggggat tctgaatctt ctgccagtaa tcccatcaaa gtatcgaaag cttcacgccc 21360
gatacgcttc ttctgaggtc ttacagaatt actatctctt gctaggtttt ctgctacctc 21420
aagactgcga ttcactgcat acttgatatc gcttgtaggg tcatacctac tcttggacgt 21480
aatgtgcgag aatctattcg tcatcgaaat cgtcctcttc atctagggaa gctgctacgc 21540
atttctcaac agctccgatt cgtattccat gattatgagc ttcttcctgc aaacttacac 21600
ttacaggcaa tccaccttca atgcgctcat acaattcatt gaagacttcc tgtttatctc 21660
taggcatgat taatatccta tgttatctat agggcaactt aattaaaaat cgggttcagt 21720
cttgtcaaat cgcaaatcac cgactttagg cagacggata acaccgttct tgctggaagg 21780
ctgcaatccg tatactcgga agatcttacc tatagggtgc acaaaaccac acggtactcc 21840
atgaatctca tggaacatac actcagcatc atggtgcgtc cagcctttgc ctagcatagc 21900
tttgactatc ttgcctccct tgtagcggaa taataggtta gcaacaagtc ctttgtattt 21960
accagttcct tcttcccaac caatgcactc taggtctact tcgtacttac gcacaacctt 22020
catttggtgc caatccttag cgccagcttt ccactcaaca tcttgtttga acacagagcc 22080
ttcatgacct aacttgatta gtcggtcagc atactgcatc aactcttcaa tgttatttac 22140
aacatcaata gagagtacgc attctgttgc agtcatctgc tgaaatagtg catcatatcg 22200
ttgcttgaac ttacgatggc tatgccctga gatgaattca caaatcgaaa cacagtcaaa 22260
gaaagcgatg tacaagttgt gcttgatgta ctgctgttct cgtgtctcaa gctctttggt 22320
tcggttaggg ttcaatacac ctgataactg ttctagcgtg tactcacagt cagttaccat 22380
ctctccgaaa tacacacctg cgtcaagacc taattcaaca agaccttcat agtaagatac 22440
cagattctct gtgtttgcga gttgctcacc agttctacca aagatgccgc acttaccatc 22500
agcacgaaca accaacatac tgaacacacc gtcacgcttg cattgagcat acattggaaa 22560
cgagacttcc ttatgcaaga taccttcttc ataattcttg ttcacctctg cgaagtgctt 22620
tactagttga acaactttgt tcttaggtcg gtaatcggct ggcttgccta ggaattcaaa 22680
tacattttgt ttgctcatgg cgtgttacct ccttctagcg taacccagaa agttacgcaa 22740
gggattaagt taatacaaat acgtttatta taagatgacc agtgcgctcc tatccagaag 22800
ctacctagtc ggcagataac cccacacttc attgtagctt agccttctct atcaatagtt 22860
caatacagtg cttagctttc tcaagctgct gcacttcgtt atctttctta cgaccaaggt 22920
atttattaac cttggtatgt atagcagcct ttagtccagt gtacccataa ttagcatatg 22980
tgaattcaag tggttgtatc cccatatcta tgtaatgact accacctact tgagtttcac 23040
tggcctttga tgtaattttc tgttcttcgt agaaatctcc atcctctctg atacatttta 23100
attgtagcgg tgttgtcctg caaccagccc tcatgcacgg gcacttagtg cacatctcac 23160
tgtcagggta tccactgcaa cttgctccga agataaaaga gctacatgat tctttattca 23220
ttataacctc ctgttcagtt gtaataagga taggctgtct cacgctgtga gacagcccat 23280
agtttattac agcttaccta accaacgacc gtccttacca agacgcattg gaaccaagat 23340
aggcaatgag tcaacaatta cagaacagcc aatgataggc ttgtgcttga agttctcacc 23400
ataagcaaat gcaagtgagc caccatcaat caagcagcca ccgtacattg cccaataaag 23460
agcagcacga gatgctgaat aatcaataga gtacttagca tggaagtgac ctactactag 23520
gttgcaacgc tcatgagctg ctgcgtctag caagttgccg ctagcttggt gctggaactg 23580
aacctcttca ccattaggta atggtagtct atgtgagtac gcccactccc aaccttcacc 23640
accaccgtta gggaacaaga cttctcggta tgacttaatc atatcagcag gaatgccatg 23700
agtcttcgct ttacgatgca gcatagagcc gtgattagag tgacagattt tcatctctgg 23760
gaacacacgc tctagttggt gtaagaattg acgagcctta actagctcca tacctgcact 23820
atctaggtta gggtcagagt catgatagct caatgcatgg tggtctgtct catcacctaa 23880
gttcacaaca gtgtctggct tgtacttagc tgcaacttca atcaagaagt cgagagtatc 23940
tgggtgatgg taaggtgcat gtaagtcagg gatgactaag atgctttcat gctcagtttc 24000
tgacacaggc ttaagaggct tgtcatctaa tggtgtaggc ttacgtagtt tacgacttag 24060
tcgtagctga cggttagcgc gtgttagtgt gatgtagaac tcaccgttct tcttcttctc 24120
ttcgtactgc attgaccagt aacgagccag ctcaggagag acttcagtac cgtgactgtt 24180
gttacttaag atagctgaca tcttcttgaa gtcaatagta ccatcgcttt tagcagattc 24240
tcgaatcgca ttgatacgct cttcatctgt aaagcggttc caaatcttag cattgcttgc 24300
cattatttat ccttcttctt taatgcataa cgcttacgtg ctttagcgtt tcgcttagca 24360
cgagcctctg ctggtgttaa atgctctgga tgaatccact cagtctgcgg aacttgatgc 24420
agctctaagt agtcagccat accgcgtagc atgttaataa tgtcacgctt gcttgtgcag 24480
cctccccatc gaatacatag gttggttagc ttaccctcaa gaccattgat tgcacgaggc 24540
aatgcagctc ggataactcc gctcttgtgg tcgtggtcaa caactacatt cgtagaagtc 24600
attgaacgga ggtcacgacc ggttataggg cacttgccgt tctgtcgtcg gataagctcc 24660
ttcttgatat tagggagatt atctgggtta tccttgtgct ttactgcacg agaactaatt 24720
gctttacgca gttctgtcat gccaaaactc cttatcattt ccgtagatga taggagattt 24780
actacgccag atttcattag ggtaggtctg catccatgca agacggcctt gctctagcat 24840
acggtcatat gctgttaggt atgctccttt ccctttccag aaatcccaat caggcggtgg 24900
tgcgttatga tgcaacatgt agttgtcata gtactcttgc gtacctctga agttagggca 24960
ccagtgctta cctgttccat aaacttcttt gtatgcacct agcacagcgt agtacatctc 25020
cttctctgtc ttacagttat caaggaggtt ataagcaaag gtcatgccct tacctttcaa 25080
gcctttgtag ttgtctgcta cgtcacccat aataatctgg gcatagaaga actttagtcc 25140
agtacctttt aaatcggtaa tggatgtgct aggccgcttg ccaacaagaa cacgtttgac 25200
cttctcctgt cctgccttag ccccacgctt ccagaactca ccagtaccta cgtactcata 25260
gtgattcact tctttattag agaacttagg attcaactca cctaatggat taacaaagat 25320
tagcttctga gtatcaggat tgtagtgcca agtaggtgtg attgtactat ctttatcaat 25380
agatatagta acagagtcac atagttcctt gtgcatctca gagcctagct caactccctg 25440
ctcttctagc aatcggtatc gtctccaagc ttcaatgctt agctcatcat ctgcttcgtt 25500
accatcagac agcattgcct taagcaagaa aatcatctgt gtcttaagct cattaaagaa 25560
aggtggttta tcttctggac gctgaccttt atagtcagtg gtgtatgcta agtctaatcg 25620
gaagttctta gcagagtcag tggcatacag cttagcagag tcgcaacctg catctctaat 25680
ccacttattc aaggtttgac aaagactctc aaatgcagac tcaaactgtg gagtatcttc 25740
aatgcatttg taatgtccac ctttgacaag ccccttagca gcaagctctt gcgcagggtc 25800
aataacaaaa gctatccggt aaggaagtag gtctgcatct actagcgcgg tacgagtgcc 25860
cgtctcaggc caaaggataa agtgctcttc cttctctgca ataccaccac catagtcaaa 25920
cccttcatag tatcctatcg tcattataac ctcctactca ggattaacaa aaataagccc 25980
taccgcatct agtgcggcag ggcaagtgac agagagttct tagaactctt cgtcttcagc 26040
tacttcttct actggtgctt cagcttctac tgcttcttct gcaacaggcg cttcttcagc 26100
agcttcagct tgctctttgt acattaggta agcagcaaca acgttagctt tagctaccat 26160
tgatttctcg gtagcacctt cagccgcagt tgcacgagcg aaggtcaaac cttctgcgta 26220
atacttaaga ccgcctagct ctgcctgagc ttctgcttct gtcttgtgag ctttcgagcc 26280
tacaagatta cctgcttcgt ctacaacaac gaattggtca acgaacttac cagctactac 26340
agtttgaagt gatttgatta ctagtttaga catgttaata ttccttagcc ttacggcagt 26400
tattaaaaga gattcagtta tttatctata gggcaactta attaatccgc cctgatttgt 26460
ctcacgccgt gagactaagc gatgtgctta acacgcttaa gagtgtggat aggatagtcg 26520
tagtttacac gcttaccttc ttcgttgata aagttaacac aagccatacc atcacgatac 26580
agtaagtgta actcgccgca gccttcacct tcgatgaagt gagcaccacg gtatttctta 26640
gcttgctctt cagttgtgat atcgttatca tcttctttag caagacgagc agcaccagct 26700
cgtgctgcgt cttctggatt cagaatgatt tctacttcgc tacgtttcat ttagaagtac 26760
cccgctactg cgccaatgaa tggaatgaat acacctactg cacgaactac agtctcgcta 26820
gttgagttag tatccatctc tgctactagg tcatagatgt taattccata acctacacct 26880
gccgcaataa ctacggcgat aataagcagc acaagaagtg ctgctcctgc tttcttagaa 26940
gtcatcttaa aactcctgtt cttcatctaa ctgctctgct ggctcttgcg gaagctgacc 27000
agcgccattg ccttgctgtt gattgccaga ctcttcttcg tcatcttctg ctttgtcttt 27060
agacttagca gtgtaacgct ctgggttctc gtcgtagatt gattgaatca actcttggca 27120
agggtgagta cctgctagga actcttctgt ttgcatcaag ataccagcaa actcacgagt 27180
aggatgcaag tagtcaagtg cttcttttgt caattggtct tcacgtagga agccaatagc 27240
gttatcaaga ggcgctagct ctgggtctgc atctagcatc ttacgcatac ggtctgtgat 27300
tgcagccatt gacttgaagt tgacgtactt aggagtacca tcttcattct tgtctttacc 27360
acctaccaag ttcacagtgg tcagctcatt aatcatagtg ccgaagcctt tatgacgagc 27420
cataccaccc attgcaggga tgaacttagc atgcaagaat gacttgtcac ctttcttaag 27480
agggaaagtt ttagtgaaga acataggctc accatcatcg agcttatcat tctcacctag 27540
taagtggaag attgccacag cctgtggagc tgctggtttc ttctgacctt tgaaagtctc 27600
ttggaatgta ccaagacgca atagaccgaa tagacgggct tcacgctcac cctcttcagg 27660
gttcttgtat ttactaacta cctcggctac ctcgccacca tagtcaaaat cagttgcaga 27720
catttgctac tccttctgtt ggatgaatta aagtatctca aagatgttgt gccagaccac 27780
acgctcacag ctaattgcct cttaggtctt gacgctgcct gtcaacatct ttgagatact 27840
gcctcattac aaggcagtac ctacatcctc tattatcttc tcttatcaaa actgcattca 27900
gatatcatgt aaccagtggt tagggtgata gcctgtctct ccgataacta gtgggagtaa 27960
ccactcatca ttgatagaaa cctcgcagga tgataagaga atgaaaagaa ataatatcat 28020
aatcattctc ctaatttata ttatctatag ggcaacttaa ttaaagctta ttaagctctt 28080
caatcgcctt gccagccgct tctttagcag cgttagcacg ttcatgacct tcgtctagct 28140
tatcagcagc tcggtcttct agcatcttct taagaacttc tgctgtttct agacgcttga 28200
gatacagttc atcttctacg tcgatacgct ctgctgctag ctccatacgt gtcacttgct 28260
tattgattgc ataggtacgt acacgctctg ctagattgct gatacgctct ttagctgcta 28320
gtaatacctt aactacttta tacatcttat actcctattt atactatacg ctctatctat 28380
agggcaactt aattataggt agcctgtgag acaaattgta cacaggctgt gagacaagtt 28440
aatacttagg cttattccaa tcatctaata gagaatagat aattaggagg aaagaaacta 28500
atagtaagag gttagtgggt gtcatgccag ctatctccta tcatgtattc acctgctaat 28560
ggtactcgga acttcaagaa ctcacctgct ttacgcatgt tatctgctag gatatgccct 28620
gcacgatgat aacatcttcg aatagtaagg agtccagttc cttttccact gacaattctt 28680
ggtgctgacc agatttgtcc gtcatcatcc acgaactgag ccttctcttc tgccttccag 28740
tcttcttctt taatctcgta aagtttctct ttaacttcat ctgaatttat ctccatctgc 28800
cattcgtcgt ggacatctgc tagccaacaa gggtatccgt tctcatctag tccaatacct 28860
tcttcaataa gttggttctc tgccatacat ttactgtact tcatggatag ggaacctgtc 28920
atctgaagca atacgttaag gacagtgtga ataagaatct taccgctctg cttacggatg 28980
cgaccccaac gaccatcaat cgcatgaagg tatccgtaac gctcacctgc cttctcacaa 29040
gctttgatta agttagcaag tgaaggtagc tctttcttga atcgagcgac tcgtttctcc 29100
atctcacgct catcaagacc acaaacctta gctaagttct taataccaga gccgtacaag 29160
aaggcataga taaatgtctt agccatatca cggataggca atccagctag gtgttggttg 29220
tgtgtatgta tatcaccgtt cagaacaatc tctgtatact cagggtcgtt catgaagtgc 29280
gcaagcatac gtagctctag acctgcaccg tcacaaccta gaatcatctt gccttttcca 29340
gcaatgaaca ggtgtctcaa cggatgcaag ccacgagaag ggatgttaac cacatactta 29400
tgacgcatac ggaaagtgtt agtaccaata ctgaaagctt cagcaggtac acgccattcc 29460
tcatcatagt cagaagggaa ctctcccatg ctctgatagt actgccacgc ttcacatcca 29520
tgctcttcat tgtaagcctt agctagcaaa cctttacagt atctacgtcc gtccttctgc 29580
ttcttccact caccgttagc ttcgaagtac tccatgtcct taacgttaag aatctgagag 29640
cgacgagaac gtagcacata ccagtcaact atatgctcta gccactgagg tatctctcca 29700
tcacgagcct tccaagcttt aagagaatcc tcatcaatct taccagacca aggcttaggt 29760
ggttcacctg tctcgttctt gtcatcgttc atccacttct catcagtctc tgagaagtta 29820
acaccaagcc aaccacgagg gtacagcacg ttctctttaa cgtactcaag gtttcctagt 29880
ccaatgtctt cgaacacaat aggagtgtag gcacctgcta ccaatgggtc ttttgtatca 29940
tgagcattgc caaccatctc tgggaaatct ttcttaacag cagcagtgta atcaccctta 30000
gctgtagtca acttccagat agttgttcgc ttacctatac gcggctctgg cttagagatg 30060
aagtagttct gtagatgact tcgtaaccaa cctacatcag cgtcagggta cgctttagcg 30120
aacgatgtac acctgttaag aatgtgactg ttcttcatag gctctgtctt gacacgtggt 30180
gggatgtaag gttcaatctt cttgaacact tcactcatct ctttgccaat ctgaacccaa 30240
tcatcccaag cttgtttcat atcaagacga aagccacgct gagcttgacg agcaatacca 30300
agagcaacct gtgtctccat acgtagagca gaatcaatac ctagtcctgt acgtggattg 30360
cgtcctcgac gaagatggtc agcccaatca ttgttcatca accagaagaa catatcctta 30420
ccaatggctg tatcctctgc acatcggtcg aacaccatga agtcagtaag ctttgaccaa 30480
tcctcattac atggcttgta tctcccgata cgaataccgt gagcctcgat gctatgagca 30540
cctacattac ccattcccac agcgaatgct tgaggtggag ccttacggtc tgggtttgtt 30600
agctgagaga taagcattgt atccatcagc ttaagaggga agtagtcaga gtgctctcgc 30660
ttcttaccac gacgctctag gtaattgaac ttccagtact taggccaaac cttctcgaat 30720
gccagtccgt catagccaac acagttctgc atgaccaagg actcacactc catcagcata 30780
tcaagagcgt caactaagta accatcttga gtaccctcac cgtctagcat ctcacgagcg 30840
cttgggtcac gcttctcata cgggtcgaag aaaacaaact cttcatcagt gaatgcatca 30900
cggatacaga tacagtgcat agagctaggg tcattataac gtagctcttt aagcaagccg 30960
acaccttccg catcggctac taagaagcga cctttgccgc tcggtttatc agggtcgccc 31020
ttcggataca tagaagggta atgttcaagg gacataccgc ctccttactt acggattatc 31080
tctgcttcac aaactggctt agcttcagac acaagcttaa ggataccaac accacagcga 31140
ttctcttttg tgaacttgga tgcagcaatc tctgccagtc gctgtgtctt gtgtggttcg 31200
cctagtactt tgccacgatg gattaacagg aactcaccaa cttcatctag tgggtctgtg 31260
ttcttatcag aaacaataga tgtgtatcca gtatctttcg catctgctaa agctttgtta 31320
tcagcgtgtg tatttcgctg cttctgctta agttcagagt gcgatacaac aaatagtcta 31380
tatttgctat tgtatccttt accattgaag acacctttca ttgtttcgat gtcatcaaca 31440
cagatatgaa agaactctag gaaccaatcc caagaatatt tctggttctt tcctttatct 31500
acaaagaaga cggtaacgtt gccaccaaga gttgagatga ttttaccagc ccaaccctga 31560
cgactacctc cgccattgtg aacaactcgc ataccattct taaggcactt gcgcatttgt 31620
ttatgaatag gtagaaattt aacagccatg atattatcct catatgatag tagaaataat 31680
caagagctgt ctcacactgt gagacaaccc tttgttatta ctactcgtta gtagttttga 31740
agtagtcgaa catcgccttc atcttacgaa ctgtcgtcaa cttagtcagg tcgttgtaag 31800
agaagtgata gtcagggaac ttgccaccat taacacggac atgcagctct cccatcaact 31860
gactatgatg ccagtcaatc ttacagtcga agtacttacc acccatttgt gctaggtctt 31920
tacgaatctt actatcagta atgatgtcta gcatataaga aggtacgctc ttaatacgta 31980
ctcggaatgt ctgagtcata atctcttcga acagtaccgc cttagcatga gcagttgcaa 32040
agtgagcaga gctacatgac actagcactt gagcaatagc gtcaccttca caagctaggt 32100
ctactacgtc atcgtagctc atgctagata gctcgtagaa acgtgccagc tcttcttgtg 32160
tcatcttatc tttgttgaac atgatattaa tctcctaatg aattatctat agggctactt 32220
aaaactcttg agtgtcgtct acatcgaatg gtacatcatc tggtaatgta ctaggctcag 32280
actctggtat cgggtcgcta cgtctggagc ggtcatcgcc catgtcgaag ttgtcacgct 32340
ccctttgtcc tgcgtcattg gattgcttac gacctacctc tggtagctca tgcactcctt 32400
ctagttcctt gtactcacca gttcgtattt ccttctcagc aactacagta ctgcctacca 32460
tgtgaccgat gcctcggttc ttaaggttac gatacagagt gatacacttg ttacggaagg 32520
tagaggcaac tgtgttacgc tcaatacccc atacagcatt cgcccagaat gtaatagagc 32580
cagcaccacg gaagtcagac tcgtacactt cgccacccat cgtgtgtgga atgcgcttgc 32640
ttgcgtcaac tttaacaagg tgactaagta gcatgatgtt gacagggttc tcatccttga 32700
aagtaccaag acgtttcata gtagcatcaa ttgcttgaac acccttattg acatttcctt 32760
tctcatcctt gtgctcaaag gcagtaaggt tatcgacaac aaagtactgg tatcctaatg 32820
cgaaacactc ttccataact tccatcacag catccacatc cttagagcct tccaagtcag 32880
cgatgatgag acggtcttgc tgctctagta attctagtgc agcgtctaag tcctgttgag 32940
tgtagtcacg agcagggttg tactcatacc cttcgtcaat ctcccattgc tcttgtggag 33000
gtgagttgaa gtccttgtta actagcatac ctgcgaaggt cttagttact tcatcaatct 33060
ggttctcaag gtagataaca acaacgtcat gacctagatt cattaagtta gatacatgag 33120
ccatagttgt atctgtctta ccgacaccag taccagctcc ccacactgac atatagtgca 33180
gtcggatacc ataggtaatc atgttgaagc ccttaagcca atacttctca cccatagtgg 33240
taatctcacg agccttatcc ttgatatcag atactcgctt gagtcttgcg ttagtctttg 33300
actcttctgc attgaataca gcagttacga actcagaacc acgccctttc tttaggcagt 33360
cattagggtc tttacaacct gctggcatta cgagacgctt tgacttccca cggaacaatt 33420
tagacgctgc cttgttcaag gctaaacctg tctcgtcgtt gtcgaatgcc cagatgatag 33480
tcttgaactg attgatatgc tccttattat caataagttc ttgcacacct gcttcacctt 33540
tattgacaga ccatacatgg aacagcttaa gaccttctaa atctttacga cctgcaaagt 33600
tctctagttt gttcagctcc ttgataagca tttgctgagc agccatagcg tcacactgtc 33660
caccagtaat gatgagcatg ttcttcatct gaccggactc tgctacattc ttcaaggtgt 33720
gcataccaaa cagctcttgc ttaccgaatc gcttacctag atgaccgaag cggaaatctt 33780
taggtatagt gcggcaagta gcgcccatca gctcaccttc ctcatactga ggatagtagt 33840
gccgacatat cttacccttc tcatcgtgcc ctactcttac gttgtagagc ttggcgataa 33900
gaccatgaat gcctcggtca accagatgct gagtcttcag accatcgaac cattgcatct 33960
ctagttccca ctcagcaagc tgagcctcac gctcttcgtc gttcaagact tcgaagcggt 34020
ctttcttacg cataccagag agcgcaagct gtcgcatgag ggggtctttc aacttgccag 34080
atgattcaag ttctttgaag tcacctaccg agtactggat atcaccagag ataggtaggt 34140
ctttaagagc aggggcagag ccatcctgct cgtagtacgt attgttatcg ctatggaact 34200
gcttacgata acagaagcca ccaccgtctg cgaatcgcat caggtggtcg cctgttttgt 34260
cgtgtcccct ttctctacac ttaggacacg gctcgttacg gattatgtca cccatatcta 34320
ctcctgtctc acactgtgag acaaattagt tacaccaacc gcgcttcatt tcacggatgc 34380
gtttacgatt acgcgatgag tttgtctcat gcgccttctt gtgattgtgc ttctttctag 34440
gaagcttctc atctacctta gacattattc accttcgaat gatagtacgt cttcttcttt 34500
gaatgtaatt actgtgccac ttgctcgtag ctctagcacg ttgtaagcag agaaggcacg 34560
atagcctttg ccattggtta ggttcacact gattaagtca tccttgtgcg cgatagtaga 34620
ctgaccacct ttcaagtctt tcttaacacc agttcggcag gtcattgtca tgaagtgctc 34680
agtgtgctct gagatttcac atgtttcaga atccttgatt gcttgaagca actgttgatt 34740
cttcttggta ttaaggtaga acttgtcacc ttcactaatc tcaacagtgc ttctgcctac 34800
tgtatagctt gcagcagttg tgccaccgag tagcttgcca ataacaaagt acttcttagg 34860
tgatttgcgc ttagcaatca cagtgaaaat cttaccaccg ttctgacgga tgatagcctt 34920
aactagctca gcgcggttct tatctttaac acgaatagtc atgatgtttc tccaatttaa 34980
ttaatctata gggcaactta ataatatccg tagaacttac ggttctctct tttgtattga 35040
agctgatgtt tctttatctg actctttacc agcaaatata aagaagctat gtctaattgc 35100
tcatctataa catgacgagt taagcataca tctctcttta tagtaagagg tgtatagata 35160
tctagtatcc atgcaccata ccaaggctta tagccaacat cataatataa ttgaataacc 35220
ataataactc cttgtctcac actgtgagac aacttaatgc atcttgttaa taagagctgt 35280
ctcaatatca gcatctaata ctgattctaa gtactcttca tataaaggat ataagtcacc 35340
ttcttctggt agctcatcct gatatagctc tagccagtca ttgaacgata gtttcttaat 35400
catagtgatt gctccttatt aatctatatg gctacttaat tatagccgct ctagtagctc 35460
ttattcttct tattctacta atgctctatt ctctctagcc gctctcctat agggcaactt 35520
aatagtatag aacgagaaaa gccctgactc aattaagaat cagggcagtt agtttaagcg 35580
aaacagtaca aggatttaag aacttcttgc aagtcaagct ctcctttctc tggtacagtt 35640
gcttctattt cagtaaggat aagagcttca ttatactcta tgaagtcttt gattacatca 35700
ttctcttcat acatctgtac gaaagaacca gaaagatgca gtctaagtaa ctcagtatta 35760
cctgcatgag taccgaaaga gtcatgaata acagcaatgc tagttattcc gttgtcctcg 35820
aaggcacaaa cagccagtac taagtggctt gcatccatac tatgaacaaa gttaggagcg 35880
gcagagctag ccattctgcg agggtctatc ttatctgtac gtactgaggt acggaagaaa 35940
gtttcaccta gcatctgtgt cttgatgcgc ttatcttctt ttacttcgta aacagcttgc 36000
ttaacgatga atccagtagg agtcatccac tcaagcggct tgttatcctt agcaatagca 36060
gagcaaactt ccttaatgta cttcatacct gctcttgcag ctactacaac ctcaccgatt 36120
gcagcccaag taagcataga agcgaaggag atagcgtcac gaagtggcaa atccccatca 36180
gaatccgtaa atgcatgaat actacctgct ttcatgcctc ttgcacgagc tttgttgtct 36240
gctttctctt gcaaatcttt aagataatca gcaatagagt cacgacaagt aagctgagag 36300
ctaccataag gtagggtcat tactggcttc ttagtcattg aacgagttac accaatgcgc 36360
aaccattctt cacagaactt gtaagccata atacgagcct tgctctcaag gtcgccagtt 36420
agcttagcct tctcgaactt agctactagt gaatcaataa gcttatcata gataggacag 36480
tcacgctcat cgttgttagt gatagcttcc atccactcga caacaacttt agatacagca 36540
ccgtatatat cttgaggctt ctcagatggt aatagattta cttccttacc acctactgaa 36600
tcccgtagca tagccgaata atgttggata ccagaacaac tcccatccat agccacagca 36660
atgtgactag gaaagtcctg ttcgttattc ccttcatcaa tccactcaag aagtgctgca 36720
tactcgtagc accaagcaag gaactgccaa ggcttatcag ctttcgtcca gtcagtgaaa 36780
gtaatcgggt cggctgcaat gtcaaggcac atctccttaa attcttctgt ctcacagcgt 36840
gagacacgtt ctgcaaatgt ttccttatcc caaccccaaa cgtttgcacc gtgtaccttg 36900
aaccagtacg ctccagtgct gcctaacggc atagattcag cgaatcggat aagagccttc 36960
tgtaaatctc cgccttgagg cgatacaagg ctagaatgac agtaaactcg accacgaaag 37020
tcaagagtat atacgaaatg taggttctcg aagtccttga acttgtttgc ttggtcaaga 37080
gtggcaacta cctcacgcac atcagactta cgcttgttct ctttagcata aattgatgca 37140
atttgacgct tccagttctt gaatgctgct ttctggtcat catcaagcac ttgcatcaag 37200
tcctcaccac gcaaatctga gtagattgca ggcactggac acgcaggttt gctatatttc 37260
ttgaactctg gcataccaag aggcagttca cgcaagcgaa tctcgtttgc tatagcgtaa 37320
attgacgcgt taatctgcca ttttacattc tgaagagcgt taagagcctg ataaacttta 37380
ggcatctgct ttctagttag gcggcgaagg tgcttcttgt ctcgtacctt agcgataggt 37440
agattagaag acacctcttt gctgtgatac ccaccattga aaggtgattt ccactccttt 37500
ggaggcacta cacaaggctc ataggcaggt gacatattac caaccacctc tttatagcct 37560
tgaatccaag tctcaagagc gtcagtagct acaatcttag ctgtagtctt accacggcct 37620
tgaccattca cttgcttagc gataagaggt tgtccgtcca tgagcatatt cttaccgaaa 37680
atctcaataa gctttgctcc tagctgaata acatcattat ctgaccaagg tatccaacgg 37740
tcaatatcaa tagcgaactc agctttatca agaagctgct gatatttgtc attatcaata 37800
gaaagaagtc tcataatctc aagcttctct tcacctgcac caagaagggc ttggaagtca 37860
gctagtaact ttagttcttt ctctgcatga accataacat catggtcaaa tttgtaactg 37920
ttactgttac gtttcttcaa gctctctttg attgcttgaa tgtatttagg tgctgcatta 37980
tttagcttag taaaacgaac ttcgtcttca atacgacgac cgatattatt agctaagttc 38040
tgagcggtta tttcatgatt agaaagacca tcaaagatag tcttgatagc aatatatgct 38100
gccgcttctg gtgacaagca cttaaggtgc attaagcaag cactaggctt accacgtcta 38160
ccagtgtagt agtcaatata tgcttgaatg gcatcagcca tagggcgtac aaattcacgg 38220
gtcagacgtc tgtaccagtc tgcatcagag ccattgccag attcaagagc gcgttggttg 38280
ttcttctcaa atctaagaat acctgcacca tgcatctcag cttcaagctc taactgaatt 38340
tgcattaagt ctttgttcat aaattactgt tccttacaat tagtgatacg tctagcctgt 38400
tgacttacgc accataatat atctttctct tcgtttgtat actcaatctt atcagcacct 38460
gagcaataag tcaaaccgtt gtgtaaccac ttctggtata actcaaagtc acgtttaatg 38520
agtaaacttt taaaagtttg ataaggaagt tcagttcgca ttgtgattct ccttgtctca 38580
cgctgtgaga ctggttttgg ttaaagcata tcataagagc cactaggata gtgactctga 38640
gttatgctct attcattcca ccatatagtt gacagtactt tcctactagg tgaataccac 38700
atatcaacat tctctgactc aacctcaatg aagttgtcac gaaagactgt acgcaatccg 38760
tgacgctttg catgtgattt aatcatacgc tttgcgcgtc ctttagtaat accgtaagaa 38820
cgggcagtga tacggccaga ttttcggcct tttaacatag ttttcatggt cttttcctta 38880
gagatagtta aagcatatca taaaggcacc tcgtaagatg ccttgagtta tactctatct 38940
gtttcgtttg ataccctgtc gtgcgtcact agctactagg aaggctggct tcttagcgtt 39000
agtagctcct ttctcaatct taggcttacg gcactcattc cactcaacct tattaagctc 39060
atccattatg atgtgctgaa aagggaatgg cttgttaccc ataaactcat tgaatgtaat 39120
tggcttaaat ctaaggttaa tacctgcaag agagttagcc atagcagcat cttgagcatg 39180
agctatattt gcaggtactc caaatgcatc tttcttaatt gcatgatgct ctttcttgca 39240
cgttttcacg aatgagacag ggtcagtaat tactagatga ccaaacttgt caaccattac 39300
attaccctcg tgcaaatcga actttgcaat acctcggaag aaatcacgca ctcggatagc 39360
ggttctgtag atacctagtg aatcgttgtt atcattgcaa tagaagatat cgcctaattt 39420
aaagaactgc ttcataaaag aaggttcttc tatagcaggc ttattagctt ctgcgtgata 39480
acgacgctta gctttataga ttctaacaag cttgtcataa gctactagtt tatccataag 39540
cacaacataa caagagctga aacgctcaag accatagata gttggtatac cttccaagtc 39600
ctgattgtca cggcagaaag cagcgtaagc cgctccactg tcttctttct tgaaccctag 39660
tttaatagcg tatttcttga tttctggatg tgtccacgct tgagagaaga agccagagcc 39720
tagcttaaca tagccttgct ctttcgctgc cttctcaaag tctcgaactt gatagtcgat 39780
ataatctccc tctcttacat cctttaccat tgaatccgct agattgatta atccacgcac 39840
gatagggttc tcgaaagtaa tcataaagat ttcctcttgt taggttggtt agtgtatatc 39900
atagaagccg ctctttgaac gacttcgagt tatacactta tttcttagct ttctcagctt 39960
caatatcggc tagtagttgg tctttagctt ttactagtag ctcaaaagct tcgtgaccat 40020
caccatatcc acatttgata agttcacgga atgccttttt caccttagta atctgcttag 40080
attcaagttg accaagccct agtactgcat aagggcatgg attcttgaac tgtggcagca 40140
taggcgcaac tggtttgcgt tctgacttat ctcgcaactc ttttagggtc ttattaagac 40200
cttcaatcgt tgctcttagt tccccgatta gcttatcttt agcaagtatc tcttcttgat 40260
taacaggtgc attctcttct tgaagcttgt cattagcttg catctgagat tgcacactat 40320
caggttcacc agtgtcaaaa ggtacgtcat ctgcatcagg aagcgagtta agagactctt 40380
cagcctcttc tctcttctct tcagcttgtt ctggtgtctc ttttgacgtt tccggctctt 40440
ctttaggtgc ttcaggatgc aagattgcat tcaaagtacc tgtattcaaa gagtcattag 40500
ccgctagctc tgccgctttc tcaagctgct cttgattagc ttcggtcgct agcacataaa 40560
gcactcgcat agctacgcca ctgaatcgtt tatcatcttc aaagaattca gatactttca 40620
tcagcttgta agcttgtgac ttgccgatac tgaaattaag cttacagtat tccaagaaat 40680
cggcttgctt cttgccttca tctttcatga tgctaagagc ctcacgcagt agattaccta 40740
ccgctaagga ttcttgttga atcgtgtcaa gttttgagca aacttcaagc gtgatttctt 40800
ggatacgtgt aattgcttca gtcataatat aatcctctta atgatggtta gtgtatatca 40860
taagagccac tacttataat gactctgagt tatacactat ttagctaacc agtacttacc 40920
ttgccagtaa gtgtaacatt cactatcctt agcctcttcc tcgctgatag tttggaaggt 40980
gccaatttcc aagatagagc cgtcactagt gattaagtgc gcgctcatat cgacagggtt 41040
tacagctagt acagcatctt gctgataatc tttaatgaat aagtcagcaa ccgcctgaat 41100
gttcttaagc ggaaccggag cgacgaaagt cagctctgtg ctcgctttct cttcgccttc 41160
ttccttccaa gagcctacac aagtctgtac aggtgcacta accaactcac caagcttttg 41220
agaagcaatg atagaattgt gcgcgttctc aacttcagtg ttagtagctc ggaatgcaga 41280
tgcgaaaatg atagtagctt tcatgataat tttccttagt tggttgatac atcctcaagc 41340
cctctcaaga gagaaagggc gagagtgata taccatccta agagaaccac atatcgcata 41400
ttccaaattg ttaaagagca cgaggcagat acagaaccag tctgcgctgg gagcagttat 41460
agtgcgctct cacttccgaa tgtgtagcca ttattacatg gcttgttttg cttgtcaagg 41520
attaatttat taaacctttc taagctagct cttaaagagc ataagccttt cgacttggta 41580
aacatcttac ttgataggct gactattgtc aacctttatt tgtaatttaa tttgattctt 41640
tatcttgcca gcctttccgc ttgaaccact tgtaaatatt aaatggtatc ttgcaaaact 41700
cttgaagcat aacaaagaga actaagaaca aatcaagtat taaacgtagc attctagtat 41760
cctccagtta gttctatctg tagtgattca gttcttgcct taaaggttaa acctttaata 41820
gtaagtgact ggatacaatc catgaacatt tcgaaagaat cgcagtatat tacgtgtttc 41880
attagcttta acctcttgtg tctagtggag taggtaaacc attaacagct tgaatgtgat 41940
aagttagtgt ttcgcttgca tcataccaga gactagtaat ctcttggagc gtcgcagcgt 42000
ctctattatc ccaatcaata ccaactgaat ccagttgagt tcgtattgtt accaacatac 42060
ctctatcaaa gtgattagaa gcttgtgact gtgttacttg catgatagta cctattgagt 42120
ctattaaaca taatgagcct atcaagtaag acgcttacca agttgttaaa gagcgaggct 42180
agtattacat agtaggctga ctattgtcaa cctcttttat tatttaattc atcaagctta 42240
atcattgcgt ccactgtact agctactggc aatatcttgc agcagccacg aggcgcactt 42300
agtttgtatc tatcttttat cgcatcttct ttaaagacac gagttgatac aagagaccaa 42360
ccttggttgt tagtaaataa tgcgtacata gtagttacct attaattaat gagcctacta 42420
tgtaacacta gccatatttt taaagagctt ttccgttagg acgcttatta agttatcgta 42480
tccttaatcg cttgtcaaca cttaatttcg cttatttcgt cttaagtgct atctagctaa 42540
aggctgaagg gctgccaagt tacaatgcta cctattaggc aactttatag ctagtgacta 42600
gcaaggatac atattattaa agagcaacca agatgccatc tgataggctg actagtgtca 42660
acctattact tactagtgta caagtaactg tgagatagtg taagtgcgaa caaaactatc 42720
tagtttgatt ctcactatac tatcccctac atgttcaacc ttcatcttag gtttactttc 42780
tagtgcactc ttaacccgtc tcttcattga cttgctagcc ttttcaaagc tgtcaaatac 42840
ttgacaagtt gagttaagaa tatctagttc gtggtcactt gttactgtgt ttagtacata 42900
tacattcatg atagtttcct taatgatatt agagagccta tcagatagca tccagattgt 42960
taaagagcgt tggtattaca ttcttagctc atatcgtgtt gctttcggta gctgactcgc 43020
tcacctgact gttgtcagta tcttgccgcg ttacgttgct catatcctct tgcgtcttgt 43080
aattttcggg tacttggcag ttctgcttcc cctaacctgt cgggaggtac tttgcaattc 43140
tgcttccctg tccgacgaga tgaatattaa acctacacgc tccaagctgt caacaataaa 43200
tttaaaagaa tttaatagta cgctccaaga tgtgaaccaa gtcacaattt agagcgtttg 43260
gttagttgct agctgagcgg tcttaaacct tctaatgata atagattatt accttttatt 43320
gtttgttctt tcttttaaag acaaacaaaa ggctagcgct agcaatagct agtacgctcc 43380
aagtaccaga ccaagagcac aaccaagtac aagctagcaa atactagtac gctccaagag 43440
ctaagcccaa gagctaccaa gagcatcaag agcggcacag cctgaacaca gaacgaaagc 43500
caagtaatac caagagcggc gtaggttgtt atgtgcaata gcaaagacat atgccgcctg 43560
cccttcccac agcctcagag agcggctgtc tcacagcgtg agactacttg accgcctgac 43620
cgctctcgcc cgtcctgagc ggacacagcg cgtcctatcc ctatagtgtt ataacataac 43680
attacgcctc agccgctcta gtgttataac ataacaattc caagtacgaa ccaagcctat 43740
ccaatagcgc accaagagcg agaaagagcg gcgctagcca ttatccatgc aatcgctcag 43800
ggataaggcc aagtccgctc tgagacgctt ttggagcccc tatgggggga ctgtcgattg 43860
cgcgagcggg agg 43873
Claims (8)
1.溶藻弧菌噬菌体vB_ValS_R15Z，保藏编号为：GDMCC No: 62170-B1。
2.含有权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z的噬菌体组合物。
3.权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z或权利要求2所述的噬菌体组合物在非疾病的诊断和治疗目的的杀灭和/或预防溶藻弧菌中的应用；所述的溶藻弧菌为Vibrio alginolyticus ATCC 17749T。
4.权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z或权利要求2所述的噬菌体组合物在制备抗溶藻弧菌的药物中的应用；所述的溶藻弧菌为Vibrio alginolyticus ATCC17749T。
5.权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z或权利要求2所述的噬菌体组合物在制备抗溶藻弧菌的水产饲料或饲料添加剂中的应用；所述的溶藻弧菌为Vibrio alginolyticus ATCC 17749T。
6.一种抗溶藻弧菌的药物，其特征在于，含有权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z或权利要求2所述的噬菌体组合物。
7.一种抗溶藻弧菌的水产饲料或饲料添加剂，其特征在于，含有权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z或权利要求2所述的噬菌体组合物。
8.一种体外防治溶藻弧菌有害感染的方法，其特征在于，利用权利要求1所述的溶藻弧菌噬菌体vB_ValS_R15Z或权利要求2所述的噬菌体组合物对溶藻弧菌的裂解作用在体外杀灭溶藻弧菌；所述的溶藻弧菌为Vibrio alginolyticus ATCC 17749T。
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| KR20210157183A (ko) * | 2020-06-19 | 2021-12-28 | 이화여자대학교 산학협력단 | 비브리오균을 감염하는 신규한 박테리오파지 vpt02 및 이의 용도 |
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| KR20210157183A (ko) * | 2020-06-19 | 2021-12-28 | 이화여자대학교 산학협력단 | 비브리오균을 감염하는 신규한 박테리오파지 vpt02 및 이의 용도 |
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