CN112708600A - 一株裂解性肺炎克雷伯菌噬菌体rdp-kp-20005及其应用 - Google Patents

一株裂解性肺炎克雷伯菌噬菌体rdp-kp-20005及其应用 Download PDF

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CN112708600A
CN112708600A CN202011599697.0A CN202011599697A CN112708600A CN 112708600 A CN112708600 A CN 112708600A CN 202011599697 A CN202011599697 A CN 202011599697A CN 112708600 A CN112708600 A CN 112708600A
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klebsiella pneumoniae
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杜新永
李先胜
王立坤
罗成盛
赵丹丹
张庆
刘玉庆
盖春云
马如霞
刘爽
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Rec Union Qingdao Bioengineering Co ltd
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Abstract

本发明公开了一株裂解性肺炎克雷伯菌噬菌体RDP‑KP‑20005，其宿主为克雷伯菌BK‑20005，该噬菌体头部为廿面体，长度为92.281nm，且立体对称，短尾，尾部长度为107.442nm，为有尾噬菌体目短尾噬菌体科，基因全长48585bp，不具有毒力基因和溶原性基因，保藏于中国微生物菌种保藏管理委员会普通微生物学中心，保藏日期为2020年12月17日，保藏编号CGMCC No.21408。该噬菌体能够有效分解肺炎克雷伯菌，可有效快速杀死裂解细菌，防控肺炎克雷伯菌的传播与增殖。

Description

一株裂解性肺炎克雷伯菌噬菌体RDP-KP-20005及其应用
技术领域
本发明涉及生物技术领域，特别是涉及一种裂解性肺炎克雷伯菌噬菌体RDP-KP-20005及其在禽病防控中的应用。
背景技术
随着集约化养禽业的发展,一些常见的急性传染病已能被人们用疫苗和药物加以控制,而一些以往认为不重要的病原或条件致病菌则逐渐上升为家禽的重要致病原，引起家禽大批发病和死亡。例如近年来报道比较多的鸡大肠杆菌病,鸡的奇异变形杆菌病等。而由肺炎克雷伯氏杆菌(Klebsiella pneumoniae)侵袭种鸡生殖和消化器官能引起种蛋带菌导致死胚，幼雏带菌或小鸡直接感染而引起腹泻和肠炎败血症死亡，本病发病率为38～46％，死亡率达32％。雏鸭、雏鸡在10～15日龄易感。
克雷伯氏杆菌是一种可引起多种动物患病的条件性致病菌。克雷伯氏菌对鸡的感染除了能水平传播外，带菌种鸡还可以通过鸡蛋垂直传给子代。该菌属于肠杆菌科,克雷伯氏菌属的成员，可致人和动物发生肺炎，肠炎，子宫炎，腹膜炎，甚至败血症。因此，随着近年来环境的变化，该菌成为人兽共患病的病原已被重视。广谱抗生素的滥用使得肺炎克雷伯菌的耐药菌株增多，从而造成了普通抗生素难以治愈肺炎克雷伯菌感染引起的疾病的严重后果，这使我们又重新对噬菌体安全有效的治疗多重耐药性的病原菌产生了兴趣。噬菌体是一种细菌病毒，在自然界中具有较高的丰度，具有宿主专一性强，不会破坏微生态环境。噬菌体侵染宿主菌后，裂解时会释放出新的噬菌体，而少量的噬菌体就能起到防治的效果。因此，噬菌体可以特异性的快速杀死裂解细菌的能力使他们在不久的将来可被当作抗菌药物。
发明内容
本发明的目的在于提供一株裂解性肺炎克雷伯菌噬菌体RDP-KP-20005，旨在解决养殖场禽类因感染肺炎克雷伯菌，发病急而治疗不及时，且因致病菌耐药性而导致用药无效等问题。
本发明的另一目的在于提供该噬菌体在治疗禽源克雷伯菌病中的应用，RDP-KP-20005具有较高的温度耐受性和较宽的酸碱耐受范围，效价较高，安全有效，有利于工业化生产。
为实现上述目的，本发明采用如下技术方案：
一株裂解性肺炎克雷伯菌噬菌体RDP-KP-20005，其宿主为肺炎克雷伯菌BK-20005，透过电镜观察，该噬菌体头部为廿面体，长度为92.281nm，且立体对称，短尾，尾部长度为107.442nm，为有尾噬菌体目短尾噬菌体科，基因全长48585bp，不具有毒力基因和溶原性基因，该噬菌体于2020年12月17日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址为：北京市朝阳区北辰西路1号院3号，保藏编号CGMCC No.21408。
进一步地，该噬菌体在pH在6-9范围内能维持较好的活性，其效价在5.9×106pfu/mL之上，在pH为7时，效价最高，适宜生长在中性环境中；在pH＝12以及pH＝2时，仍有噬菌体存活，该噬菌体的酸碱耐受范围宽。
进一步地，该噬菌体的最适温度为37℃，效价最高达到1010pfu/mL，在70℃的条件下作用60min，效价维持在105pfu/mL，依然具有生物活性，对温度有较好的耐受性。
本发明还提供了裂解性肺炎克雷伯菌噬菌体RDP-KP-20005在制备预防和或治疗禽源肺炎克雷伯菌引起的疾病的药物中的用途。
本发明还提供了裂解性肺炎克雷伯菌噬菌体RDP-KP-20005在制备预防和或治疗禽源肺炎克雷伯菌引起的疾病的饲料中的用途。
本发明还提供了一种用于防治肺炎克雷伯菌的杀菌组合物，包括有效量的肺炎克雷伯菌噬菌体RDP-KP-20005。
进一步地，还包括用于抗肺炎克雷伯菌的抗生素。
本发明还提供了一种预防和或治疗肺炎克雷伯菌引起的疾病的添加剂，其有效成分包括肺炎克雷伯菌噬菌体RDP-KP-20005。
本发明还提供了一种清洁剂，其成分包括肺炎克雷伯菌噬菌体RDP-KP-20005。
本发明还提供了一种消毒剂，其成分包括肺炎克雷伯菌噬菌体RDP-KP-20005。
本发明的有益效果体现在：
(1)本发明发现并分离了一种肺炎克雷伯氏菌BK-20005，并以该克雷伯氏菌BK-20005为宿主分离得到了噬菌体RDP-KP-20005，该噬菌体RDP-KP-20005对养殖环境中肺炎克雷伯氏菌具有强裂解效用，为工业化生产噬菌体用于养殖环境中肺炎克雷伯氏菌的防治提供了噬菌体来源。
(2)本发明所提供的肺炎克雷伯菌噬菌体RDP-KP-20005在治疗感染肺炎克雷伯菌后的疾病具有很好的优势，效价较高，防治效果好。
(3)本发明所提供的肺炎克雷伯菌噬菌体RDP-KP-20005在防控禽类致病性变形杆菌中具有良好的应用前景。
(4)通过对本发明分离得到的噬菌体的生物学特性研究，为下一步研发新的抑菌药物提供理论指导意义。
附图说明
本发明上述的以及其他的特征、性质和优势将通过下面结合附图和实施例的描述而变的更加明显：
图1为本发明噬菌体RDP-KP-20005的电镜图；
图2为本发明噬菌体RDP-KP-20005一步生产曲线的示意图；
图3为本发明噬菌体RDP-KP-20005酸碱稳定性实验的示意图；
图4为本发明噬菌体RDP-KP-20005热稳定性实验的示意图；
具体实施方式
下面结合附图和实施例对本发明作进一步说明。以下实施例将有助于本领域的技术人员进一步理解本发明，但不以任何形式限制本发明。应当指出的是，对本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进。这些都属于本发明的保护范围。
实施例1肺炎克雷伯菌BK-20005的分离与保存
本宿主从青岛即墨田横的一养殖场采样，无菌操作取病鸡肝脏，选用法国科马嘉尿道菌群定位显色培养基进行细菌划线分离。于37℃培养18-24h后，在培养基上显示金属蓝色带有红色晕环的菌落，挑取典型菌落继续划线纯化3次，然后挑取单菌落接种于5mL LB肉汤中，37℃、200rpm振荡培养8h，得到均匀浑浊的细菌悬浮液，肺炎克雷伯菌是革兰氏阴性的杆菌(0.5-0.8μm×1-2μm)，无芽胞，无鞭毛，卵圆形短杆状。
将分离菌接种于各种生化试验培养基中，置于37℃培养24～96小时，观察其生化反应结果，生化结果显示其蕈糖+、蔗糖+、赖氨酸脱羧酶+、乳糖+、尿素酶+、靛基质试验-、七叶苷+、丙二酸盐+、鸟氨酸脱羧酶-、山梨醇+、OF+、水杨素+、枸橼酸盐-、肌醇+、精氨酸-、棉子糖+、β半乳糖+、甘露醇+，初步判定为肺炎克雷伯氏菌。
对分离菌按照CLSI标准，进行进一步药敏试验，结果表明其米诺环素21mm、头孢他啶15mm，而对庆大霉素、青霉素G、四环素、头孢噻吩、氟苯尼考、替米考星、克林霉素等均耐药，表明该菌对抗菌药物的耐药性高，呈现强耐药性。
对分离菌进行多次纯化后，挑取单菌落置于5mL液体LB中培养6～8h，然后提取DNA，采用现有的16S引物进行PCR反应，并将PCR产物送往华大基因进行测序，结果显示，该分离菌为肺炎克雷伯菌，对其进行编号BK-20005，完成后于-80℃保存
实施例2噬菌体RDP-KP-20005的分离及鉴定
(1)本发明试验用污水样品为发病养殖场周边污水中采集，作为噬菌体分离的水样。
(2)分离：取污水10毫升，10000rpm离心5分钟，出去较大杂质及大部分细菌后，再用0.22微米微孔滤膜过滤除菌。取3毫升滤液和3毫升宿主菌菌悬液，一起加入20毫升高压灭菌后的LB肉汤中，然后置37℃培养箱中，过夜培养。培养后取5毫升，10000rpm离心5分钟，再用0.22微米微孔滤器过滤除菌，滤液即为拟含有噬菌体的原液。然后，采用双平板法鉴定噬菌体。
(3)纯化：在有噬菌斑的双层平板上挑取单个独立、形态均匀、清晰透明的噬菌斑，置1毫升生理盐水中，浸泡10分钟后，取清液，用0.22微米微孔过滤器过滤，然后将滤液适当稀释(铺双平板时有单个噬菌斑)后，与宿主菌悬液铺双层平板，此步骤重复3～4次，待双平板上各噬菌斑大小、形态和清晰度一致时即得到纯化的噬菌体。将纯化好的噬菌体铺双层平板，然后取单个噬菌斑，加入LB肉汤中，同时加入宿主菌悬液，将噬菌体增殖后，再将增殖液10000rpm离心5分钟后再用0.22微米微孔过滤器过滤，将滤液做10倍梯度稀释至108倍，取最后3个稀释度的噬菌体稀释液0.1毫升和宿主菌悬液0.1毫升，铺双层平板，测定噬菌体效价。噬菌体效价(pfu/mL)＝噬菌斑数*稀释倍数÷0.1
结果表明，其噬菌斑形态、大小和清晰度一致，有典型的裂解性噬菌体特征。该噬菌体的效价可达1010pfu/mL以上。
实施例3噬菌体的电镜观察
运用磷钨酸负染法：在石蜡片上滴噬菌体液(效价1010pfu/m L)100μL，将铜网有膜的一面置于噬菌体液滴上，10min后取下，在空气中自然干燥2-3min。然后在铜网上滴一滴2％的磷钨酸(PTA)水溶液进行染色，10min后取下，在空气中干燥10～15min，用电子显微镜观察，选取清晰的噬菌体图像进行拍照。
如图1所示，噬菌体RDP-KP-20005头部为廿面体，长度为92.281nm，且立体对称，短尾，尾部长度为107.442nm，根据国际病毒分类学组织病毒分类第九次报告，该噬菌体归类为有尾噬菌体目短尾噬菌体科。
实施例4噬菌体基因组测序
通过对单株噬菌体的富集培养后，在4℃条件下8000g离心15分钟，加入10％PEG8000和0.5M NaCl静置过夜，再加入等量氯仿混匀，静置分层后5000g下离心10min，去处氯仿层和PEG层后，加入限制性核酸内切酶消化处理，并采用梯度密度氯化铯条件下悬浮噬菌体，后期采用TM缓冲液透析3次，每次30min，最后将透析好的噬菌体一部分送华大基因生物测序公司进行全基因组测序。测序后进行序列比对，结果表明与已发表的KP1801株相似率仅为91％。
实施例5裂解谱试验
通过点板试验，将受试菌(肺炎克雷伯菌)培养至OD600约为0.5，取出400μL铺双层平板，室温静置5min，将3μL噬菌体浓缩液滴于双层平板上，室温静置至噬菌体浓缩液被吸收后，倒置于37℃温箱中12h后观察有无噬菌斑产生。分别将现有噬菌体与本发明噬菌体进行裂解试验，结果见表1。
表1裂解谱实验
Figure BDA0002870695430000061
Figure BDA0002870695430000071
由表1可以看出，显示本发明分离的噬菌体对肺炎克雷伯菌的裂解率能达到90％以上。表现出了强裂解性。
实施例6一步生长曲线的测定
将宿主菌培养至对数期，按照噬菌体浓度(PFU/mL)/细菌浓度(CFU/mL)为0.01、0.1、1、10、100的比例混合，37℃孵育5min后，8 000r/min离心5min弃去上清，去除未吸附的噬菌体，用5mL液体LB将沉淀重悬，37℃振荡培养2h，12 000r/min离心过滤取上清，利用双层平板法测定噬菌体效价并统计结果。在最佳MOI条件下，将噬菌体和宿主菌混合，总体积20mL，37℃温孵5min，12 000r/min离心30s，弃去上清；加入适量LB(37℃)培养基，12 000r/min离心1min后弃去上清，重复3次，以除去未吸附的宿主菌噬菌体；加入20mL液体LB培养基，37℃振荡培养，从0时刻起每隔10min取样，4℃、12 000r/min离心10min，收集上清液，测定噬菌体的滴度，试验重复3次。以感染时间(t/min)为横坐标，噬菌体滴度(PFU/mL)为纵坐标，绘制一步生长曲线。
由图2可以看出，噬菌体感染宿主菌后的30min内效价没有明显变化，表明其潜伏期约30min，感染后40-110min内噬菌体的效价明显上升，之后趋于稳定，表明噬菌体裂解期约为70min。通过计算，噬菌体裂解量约为172PFU/感染细胞，表明噬菌体具有极强的裂解和复制能力。
实施例7pH稳定性测定
用稀盐酸和稀NaOH溶液调节生理盐水的酸碱度，配成PH值为2、3、4、5、6、7、8、9、10、11、12的缓冲液，再将配置好的缓冲溶液将噬菌体稀释成1*1010pfu/mL，将稀释液在37℃下，水浴1h，用生理盐水10倍比稀释后铺双平板。37℃倒置培养4-6h后计数。
由图3可知，噬菌体RDP-KP-20004在pH 6-9范围内能维持较好的活性，其效价在5.9×106pfu/mL之上，在pH为7时，效价最高，表明该噬菌体适宜生长在中性环境中；在pH＝12以及pH＝2时，仍有噬菌体存活，该噬菌体的酸碱耐受范围宽。
实施例8热稳定性实验
分别对噬菌体RDP-KP-20005与KP-LYY15(现有已知的肺炎克雷伯菌噬菌体)进行热稳定性试验。将噬菌体原液分装到EP管中，分别在50℃、60℃、70℃、80℃的条件下孵育不同时间，然后用生理盐水10倍比稀释后，铺双平板计数。
由图4可知，噬菌体在70℃孵育60min后，效价能保持在105pfu/mL以上，并且在80℃时能检测到活体，说明在短时间内噬菌体能够耐受较高的温度。
应当指出的是，具体实施方式只是本发明比较由代表性的例子，显然本发明的技术方案不限于上述实施例，还可以有很多变形。本领域的普通技术人员，以本发明所明确公开的或根据文件的书面描述毫无异议的得到的，均应认为是本专利所要保护的范围。
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<110> 瑞科盟（青岛）生物工程有限公司
<120> 一株裂解性肺炎克雷伯菌噬菌体RDP-KP-20005及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 48585
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
atgttcggac tgaataagcg cagtacaacg ccgtgcgcat agcaaagcag atggccgcag 60
aacaaagacg ccatcaagaa ggacaagaag acctacgatc aggtcgccgc gaagatgatc 120
gataaacatt gggcgcaaat caacacgctg gttactcgcg gccagtttat ctggatagct 180
ggctacctgg aaggtcgatt cggtcgcagg gatggcgagt atgaataaaa aacagccaaa 240
cgattcagcg acctattgac gcgctcgcgc tgaccgggta atgtgaacca cgtagacaca 300
aggggcggta aacatccgca agtcttgccc cgcttagggg catttacaaa ggggttatga 360
tgcagtataa aatcatactc acggcaagaa aaatgggcgg cttttgcaag tcctgcattc 420
aagagttcag catgacgatt gaggcgaacg acgccgccga cgcggtggag aaaaaaaaag 480
caatccggcg ttaatctgga tacgcataaa atcaacatca actacataag ggaagtaaat 540
caatgttaac tctgattatt gcattgttaa tgctgttcat aggctatcac gttggcgcgg 600
ctcatcttgt cgagcgcctt tcaaagcggg tacatgaagg cacatttgct gccatgattt 660
acaataagaa aactgcacgt tgggaaaaaa ttggcgatcc ggatggtatc gcaaaacgaa 720
ttgccttttc gccgcttccg tatgttgact gcgagccttt cgtgaaactc cagaagacgc 780
tgaatcggcg aaacaaactg atatgacaaa agaacccgct tcggcgggtt tttttatgcc 840
cgcaatctgg tatactcgca aatcaacata gaaggaggat gtaaaatgtc tgaagaacgt 900
agaaaacgcg taacgaaatc gcactttgca ggcaacttta aagcgctata tgaaaaggag 960
ttcggcgtag tgctggggcg cacggcagag atgacgccgg aacaattttt tgatatcgca 1020
aagcgttact tccagtgggc cgaagagaac gcaatcaagg ctgcggaaac ggcaacgttt 1080
cagggcgacg ttaacgagtg gggagtgaac aagcctcgca ttttcacgat cacagggtta 1140
agcctgtttt gcggcgtgaa ccagtcaacg cttgggcgct accgtcacga tcccaactat 1200
gctcccgtca tggagttcat cgactccgtg atttatgagc agaaatttca gcttgctgcc 1260
gtcggcatga ttaacgcttc tttcgtcggt aaagagatgg ggatcgacaa gcctccagtg 1320
ctgaacatcg acgctattgc cggggataag aacgagatca ccgaagagaa gttagagaag 1380
gcagtgacca atattcttga taagctgtaa gggtcagata tgaacgaaat gatcatttgg 1440
gaagacctgt cgccagccga taagctggca attaaggcgc tgagtacgcg caacttttcg 1500
ctattcctga agatctggtt ccagatcatc cagggtgaaa agctaatgtg gaactggcat 1560
cactcctact tttgccacac ggttgatgaa attatcgccg ggaagcgcaa gagcacgatc 1620
gttaacgttg cgccaggctc cacaaagacg gaggtgtttt caatccacct cgcgccgtat 1680
gcgtatctca agtgccggaa ggttcgaaac cttcagatct cgcagggtga cgccctgtca 1740
aaaggcaact ctgatcgcgt gattaagatc ttctcatcga gcgagtggca ggagctatgg 1800
ccatcaaagt tcgggcgcaa gcagatcgat gaatttcagg tgatggatga taacgaccgc 1860
gtaaggctgg aaatggtttc ccgttcgtct ggtggccaaa tcgtcggtaa gcgtggcggg 1920
tacatgacgc cagggtttag cggccttatc gcgctggatg atatcgacaa gccggatgat 1980
atgttctcga aggtgaagcg tgagaaaaac cacgtgcttc tgaagaacac cattcgatcc 2040
cgtcgagcga agaagaagaa gggcgacgaa actccaatcc tttccgtgca gcagcgattg 2100
cacgcgcagg atgctacctg gttcatgatg agcggcggca tggccatcga ctcgatcgga 2160
ttgttatccc ggcgatggta acgcgggaat atggcgaatc actacctgac tggttgcgac 2220
ctgagttcga acgcgacgtg ctttccggcc cgtcggtggt cattgacggc gtggaatact 2280
ggtcattttg ggaggagaac gaatcaatcg agaacctggt tgcgctacgc gaagccgatc 2340
tttatacgtt cctttcgcag tatcagcagg agccaatcgc cctgggtggt aacgtgttca 2400
agtcggagtg gtggcgctat tacggcgatt ccgacaaggc gcacgagcct cgcccggaca 2460
agttcgaata tacgttcatc acggcggaca ccgcgcagaa ggtcaaggag ctaaacgact 2520
actctgtaat gtgctattgg ggcaagtacc gggatcgcgt ctacttcatt gacggtattc 2580
gcggaaagtg ggaagcgcca gatctccgcg ttcaggccga agcattcatc aagcagtgct 2640
ggcgtcggaa taaggagtgc gggaaccttc gccggatcta catcgaagac aaggcgagcg 2700
gtacgggtct aatccaggat ttaacgaagg cggtaaacgg catgggcgag atcgtccctg 2760
tgcagcgcga taaagataag gtcactcgcg ctatggatgc acaaccaatc atcaagggtg 2820
ggcgtgtcgt gctgccggac aatcacccat tcgttgcaga gcttgaggcg gagatgagcg 2880
cgtttacata tgacgattct catccacacg atgatatttg cgacaacgtg tttgacgccg 2940
caaacctgga aatgaacctg agcgacgatc cggttgagcg aatgaaacgc cttgcgggat 3000
tgaaaaagct gggtcgctaa tacataatgt gggcctgaag gcccacactt aaacaaggtt 3060
gaaatatgaa taacattaag atggacgact ataatcaaat ctttaatggt ggcgctggct 3120
atgcgtcaac cctcgcgtct atcgcggcaa gatttggaac aatgtcgcag gttgaagagt 3180
tctatcatga aaacggcatg gcgaagaaaa tcgttgacgt gatcccggaa gaggtggtcg 3240
ctcccggctt ccagctaaac ggcatttcag ataacaccaa gtttcaatca gaatgggacg 3300
ggttaaatct ggagccgcaa atcaccgatg ctctttgctg ggcgcggctg tatggtggct 3360
cctacgtcct ggcgatggtt aacgatggtc gcgcgttgac ttcggcggcg aagcggggta 3420
agccgctcga atcgatcgtt gtttacgacc atgattccgt ttccgtagca gaggaggaaa 3480
ccagcccacg aagcccacga ttcggaaagc ctaaaatgta cgaggtgaag ccgctaaacg 3540
gtgggcaacc gttcaaggtg cattataccc gtatgcacta catcgacggc gagcgagtaa 3600
ccaacaaggt gcgcaagcta aataatggcg ctggtggttc ggtgctgaac aagtcgatca 3660
ttgaagcgat tcttgactac gactattcgg aatatctggc aacgcagcta ctgaagcgca 3720
agcagcaggg cgtatggaag gcgaaaggcc tggcgctaat ctgcgacgac aaggaaggcg 3780
agtacgccgc ccggttgcgc atggcgcagg ttgatgctaa ttccggcgtc ggaaacacga 3840
tcggcattga tgcggacgac gaagagtaca acgttatcaa ctccgatatt tcaggcatcc 3900
cggaattcct ttccgcgaaa atggatcgga ttgtcgccct gtcaggaatt cacgagatcg 3960
tgctaaaaaa caaaaacact ggtggcgtaa gcgcaagcca gaacacggcg ctacagacgt 4020
tctacaaact ggttgaccgc aagcgcaatg acgattacaa gccgctgtta gaattcctgt 4080
tacagttcat cgtaacggag gaggaataca gcgtcgagtt cgaaccgttg tcgctgccta 4140
ccgatgcgga gaaagcggat atcttccaga agaacgccaa cgcggctcgc ggcctcgtta 4200
ccgaccaagt tattgacgcc aacgaagcgc gtgatactct gtcggcgtta attccagaac 4260
tgaagctaaa aggcaacgcg ccgaaacaga aaaaacttcc tgatcgcacc actggttcag 4320
gcagcacgca aagcgcagag atcttaaaca acacggagac ggatgatgaa agttaacggc 4380
agaatcccaa actggcgtta tcctgaagca agcgagcggg agttatcccg ctccatgcag 4440
gacgcggtga cagaactcgt ggtagaaatg cgggatcgct tagaccgact gaagtttgat 4500
gccacggcgg aggaaatcag ccaggcggaa gacgatatca gcgaatcggc catcgtgttc 4560
ttttccgccg taattgcggc tcttgcctcc attgggttga ccatctatag attcaactcg 4620
aaacagtggc ttgcaattgc gatcgcggct ggcgggcgga ataacgaatc cgtcatgcgc 4680
cttaaagaat tcggcgctgg cgggtatgaa gactggtatc aggaagcgct gaaaaagtgg 4740
caggattccg ccgaagcgtc aatcaggaaa ctagcaagcg atatcgttgc tgactggacg 4800
acgaaagtta gaaccgccaa caacattggc aagtctcgcc agcagatcga tgaaatcatc 4860
gaaggccgat acgccattta tggtagttgg tcgcgcaacc gggcaagcgg aatcatcgga 4920
acttttaaca gtatgttgat gatgcagcgc ctaaaagatg ctaaagtatc gcattacttt 4980
tggttcggga tgatggacga ccgcgagcgc gagagccata tcaagctaga aggtaagcga 5040
cgccccgtta atggtgacgg cattttcccc ggtgaagagt acggttgccg ttgttgggcg 5100
gttccagatt ttaacaatgt agaggtatca tgatgaaaag agttcaaagg ttcgacacgg 5160
taaagatgaa ggcccgattc gatgagaacg gctttctggt tgatactccg atcgtggcgc 5220
gtatcggtgc gcagacgtac cagacgccaa ccgggccgcg cgtcgagttc cgcccgcgtt 5280
cggaagtgtt tgatgctgaa tcactggctt cataccaggg caagccgatc actttgggtc 5340
acaagatggt gaacgcgcag aacgcaaaag gcctggtcgt tggctcgtgt tccggcgctg 5400
gcaaagaaga ggggatcggc gttcttgttc cggtgatgat ttacgacggc gagtcaatcg 5460
agcaagccaa aaagcgcgta gcggctgagt tatccgtggg ctacacttcg gttgatatcg 5520
atcgcaaagg ttggggtaat aacgcaactg gcgaatatta tttcgatgaa gaccttccgg 5580
aaaacttcga agagatgaaa aatgattccg tctctgattg ggttcgcttt gatgcggtgc 5640
aaacgaagat tcgcgtgaat cacgtcgcgc ttgttttccg tggtcgtgcc gggattgcga 5700
aattaaatct tgatagcgaa caagaattcc cctatgatga cgactcaaac cacaaaggag 5760
ctaaaacaat gatcattaaa attgacggcg tagatgttga agtggccgat aacgtaggcg 5820
cttacattgc caaactagac gcgcaagttg catcagcaac cagtcaggta actagcatca 5880
ccgcagagcg tgacgcgctt caggccaaag ttgatggcat tgaagatgaa gttgccgccc 5940
gcgttgctaa aatcaaagcc gacgaagacg caaaacagaa agttatcgcg gtggtttctg 6000
ccgctggcgt caagtgcgac ggcctggatg ttaaggcgat gaaggttgct tacatcaaag 6060
aggtagacgg tcgtgatctg tctgacaaag aagattcgta catcgacgct tcttttgact 6120
ttatcgccaa ctctgataag atggctggca atcgctccaa agtcttcggc aaaaaagaag 6180
atggcgagca aaaagacaaa ggcggcttac cgaaacttga cggcaccgaa atcatcgatc 6240
cgcaggcaaa attccgccgc taataatttg cggccttcgg gccgctacca gaacaaataa 6300
acaggagatt caaaatggca cagattccag cttcttattc ccgcaagcgc gatattgccg 6360
tagcggggca gatcgctgat acgtcgcttt ataatatcga cggcacttgt gttgctgaag 6420
gtgatatcaa agctggcgta ctggtggctt ccactggcgc agtttccaat ggccacaagg 6480
tcgttaaacc agcaaccgca gcgagcgacg tcattgtagg catcgcgcag ttctcgcaag 6540
cctactcgcc tgaaggcaag tatgacgatg agagcgcagt taacgttatg actcatggcc 6600
gcatttgggc gatcgctgac gcaactgtta cagaagcaga ttgcgcgttc ggttctttcg 6660
ttacctttac cgcaactggc accgtagcga agggtgatgc cggggttatc aaaactagct 6720
acaaacacac tggcgaatac accaaaaacg cagatggcac tgttctggtg aaagttcagg 6780
tgcttcaggg cgcggtggct ccggcggctg cggctggcgg cggcgctggt gcataataaa 6840
tggtggggct tcggccccat ttttgcacca gaaaaaaaac tttgacggct taacgattcg 6900
tgatattctt catctcgtta agctaaatac acaaacagga gttttcagat gactatgaaa 6960
ttagatgcat tcgaacagag tgccattaag gtcgcaatgc agggcatggg cgtcgacgcc 7020
gcaaaactgg atgcttacgg tatctggact gttaagcaga tgactcaatt actgaatcgc 7080
cagtatgagc aggcataccc gcagaccagc gcacttgagc ttttcccggt aactaccgag 7140
atctcgccaa ccgcccgccg ctttgagtgg ctcgaattcg atggcgtaac ttctgcgaaa 7200
attatcgccg attacaccga cgatctgccg accgttgaag cgatggcgaa agagaagtca 7260
gggaaagttt tccgcctggg taacgcgtgg tttatttcca tcgacgaaat taaaactggc 7320
gcggcgctgg gttccagcct gagcgatcgc aaggcaactc tggcccgcga aggtcatgag 7380
acgctcgtta atgatctggt gttcaaaggc tccgctcctc acggcatcgt gagcgttttc 7440
gaccatccga acattaaccg catgaccgcc agcgcggctt ggggcgatga cgcagcagcg 7500
gctgaaaagg cattcgaaga tctggaagac ctgctaaaca tgatcgaaga aactacgctg 7560
ggccgccatc acgcgaccaa catcgtaatt cctccgtcta agcgtcgtct tctgacgaaa 7620
aagatgccgg acactagcgg cgactcttat ctgacctggt tcaccaagaa ccacccgaac 7680
atcaccatta cggcgatggc ggagctggaa gatattgacg gcgcaggcac caaaggcgtg 7740
ctggcatacg aaaaagaccc aatgaacatg agcatcgaga tccctgagcg gttcaacatg 7800
ctgccgatgc agccgaaaga cctgcatttc aaagttccgt gcacctccaa atgcactggt 7860
ctcatcgttt accgcccgct gactattgcg attctcaccg ggatttaatc aaaaagcgcc 7920
ttcgggcgct tttttattgc attgcatcct acaatgtgct ttaatttgaa acctaaagta 7980
aaccaatgga gcattaacaa tggccagtaa aaaagaaacc gtagaaaccg tcgaaaccgt 8040
agaaaccacc agtgccgaac aggcgtcgca ggctgttcaa ctacaaaacg ttggtgcatg 8100
tgcaattcgc tataaaggca aaaattacgt ctatgagcag gtttttgaag tgccggaaaa 8160
cgaaatcgac cgcttccgcc acgaaatctt caaaggccgt gtcgaattct acgacaaccc 8220
gaaacgcacg cgcgaataca tcgcagcagt gaaggcgaaa gcgaaagaga tcgtgcagcc 8280
taaaagcgcg gaataacaaa aaccaacaaa gggcgcttcg gcgtcctttt tcatatcagg 8340
agatcgacaa tgaattacac gattcaagat gtgatcgaca agatgcgcag ccttgcgcct 8400
ccgcttaaaa aagtacccga cgaactgcta accgcatggg tcgtgcttgc cgaggagttc 8460
gtttgcaaat caaggttcgg ggattccatt gttacagcga tcgcacttat gaccatgcat 8520
ctaatgtttt tggatggcgc aatgaagcaa gaaggcgaaa gcctggagtc ttacacgcag 8580
cgagtggcat cgttcaccct gaccggggag ttttcccaaa cattcgatcg cgtatcggcg 8640
tcaagcgaca acgaaatgct ttctacgccg tggggcaaaa tgtactggcg tatgctcaag 8700
atgcgcggcg gcggcttcgg ccttcttacc gccgggaacg ttcgccgctg cggaattggt 8760
aggtgatcgc tatgaactac aaacaaatcc aggcacgcgc aagcgcgggc attaagtttt 8820
tcagtgacgc tgacggcgta ttcaacaagt acacgaaagg cgctggcggc ggaatcgatc 8880
cggaaaccgg ggaagatatc attcctggtg aggtggtaac gacaatcaag ggcgcgatca 8940
gggatgtaaa tgaccgtgac attaacggcg aaaccatcct cgctggcgac aagcgcgggt 9000
tcttcactca cgatgtgcca atcatggagg gtgacgaaat cgaagtagac ggcgagcgct 9060
accgcgtggt taatgcccgt ccggtaaaac caacgggaac cgttgtcgcc taccgtccag 9120
ttttacgcag ggtggcgact tatggctaat tatacgatcc gggagttcac aggggctatt 9180
gatgcatggt gtaaggccgc tggtgatgcg ctggaggacg ttgtaaggtt tacgtgtgaa 9240
gatattcacc gcgaccttgt aatgcgttct ccggtggata caggtcgctt ccgtggtaac 9300
tggcaaatca cctttaaccg tgcagcgctt tacgcgatta acgcatatga ccaaactggc 9360
gaaaagacga tccagaacgg caaagccaac attgcactat acgcaaaagg ggctgggatc 9420
acttcgatct ggttcagtaa catgctaatc tatgcgaacg cgctggaata cggccattca 9480
aagcaggctc ccaatggcgt tatgggcgtt gttgcgatcc ggttgggcgt ttatgttact 9540
gaagcaatca agcgagcgag ggcgaaaaat gcattatgat atggcgttaa aatgcaaggc 9600
ggcagtggct aaatttgccg ccgagaacgg gttgaggttg gcgggggata acgttgactt 9660
tatcccaccg agaggcgggg aaacctacct taaagcctcc tacgtcgagg cggattcaag 9720
atcggttgac ctgtcaagga aatgccgagt ctatctggcg atggttcaga ttgacgttat 9780
ctttaagcct gggatcggaa ccgaccgcgc gaggattatc gcccaacgcg ttgcaaaatc 9840
cttccctgaa ggaaagattg ttgatcgtga cggtaagtta tatgtgagcg agtgggcgga 9900
ggtacacggc gtgcagaagc atgaggctgg ttggttcttt ccggttcggt tcacggtaag 9960
atgcgagagc attgaggaaa acggttatcc atcaacctga cgggtcttag aggtgcttat 10020
aatttcttgc cagcctggaa atatataggc ataatggcgt tgttaaactt tcatcaaaac 10080
aggagtattc aacatgcatt taccaaacgg tgcaaaggtc ttctttgaga aggcccgcgg 10140
tgcggaggtt ccgtttaccg caatgaccaa cgacgcgaaa aacccaacaa tcacggtggc 10200
agatggtgcg ctcgacgtta acgatatcgt aatctttacc gattgcactt ggagcgactt 10260
tgtaaataaa gtggcccgcg tcaagtcggt atcggctggt gtggcaacgc tggaagagtt 10320
cgacacctcc gacactaaca agtatcctgg cggcgcggcc actggtagcg tgagcgtgat 10380
ctctgattgg gtcgaattgc cttgcattca ggatttaggc aaagacggca acgaacagca 10440
gttctataac tatcagtgcc ttggcgatga gcgcgagcaa tccgaaccta cttacaagtc 10500
ggcggtgacg cttaactaca cgtttgcgca cgattacagt aacgcgatct accctgtgtt 10560
gcgttcagcc gacgccagca agcaggcgaa agcgatgtac atgtatatcc cgcgagcttc 10620
cgaggttcgt tactggtctg gaattgcatc tttcgatgac attccttcca cggctgttaa 10680
cgagatggaa acggtaacgc ttaacattgc gcttaaaggt gcgcacgtct tccttccggt 10740
tgtcgcgtaa ttaaatggcg gggcttgtgc ctcgcctttt tttgtgcata atagcgaata 10800
acacaaacca atcaggagtt aacaaaatgg ctaaattcaa aattcaaatt ggcggcaacc 10860
tcccttcttt caagctgccc gtaaccttca cttgcccgga tggcaaagaa gcaaccatca 10920
ccatgaccgt aaaacaccgc tccaccgatg agatgaaaga cttctatgag agcgaagata 10980
aagcgccaaa gggtaacgcc gagtttatcc gcttcatggc tgaaggttgg gatctggatg 11040
acgaattcag cgatgaaaac atttcctggc tttgcgctca cttcccggca tttgtcatgg 11100
cactgccaca aacttacatg gccgcacttg cgggccaccg tgcaaaagtt taaggcgggc 11160
tgtttatctc acgcttcagc ctgagctaac cgatcgccag cttgcggagt acgggttaag 11220
gcgatcggac tatgaagcag atcttgaaac gatatatttt gatgagcaga ccgcccaaag 11280
ctggcagcta ttccaggcca tgcaaacgca atggcgaatc gggatgaatg gcccgacggg 11340
gcttgactat aatacgttgc cactgctgtt cgaattgtat aaaatcgaca atcgagaagc 11400
ggcattactt gacttgcaaa tcttagaggg tgaatacctg aaggagattt acaagaaatc 11460
caaataagcg cctacgggcg cttttttcat atgggggcta aacatggctg ataaagtagc 11520
tgggttgacg tttggcgttg acgtttcgca ggttgacaaa gcggtacgat cactcgcaga 11580
actgaaaaac caaagccagc aaacgggcgc tggcctacag tcacttgcgg acgctgaaag 11640
gcgggccacg gcgcagaccg aggaaatgaa ccgcgcgttg cagcgccaga agcaagagac 11700
ggacaaatca aaaaccagct ttagcaggat cgcaagcgcc atcgatccca cgatctcaaa 11760
aatggccaac ttgcgcaaag cgacggaaga acttgataaa gcgtgggctt tggggcttgt 11820
tccagataag gaatttttcc gccttggtgc tatcatcgaa tctactacca accaactccg 11880
gcggcagcag gcggcgctaa ccgaagaagg tcgcgcagca atcgcagagg cggaggcaaa 11940
gcagaaggcg gcaaacgccg ggcgtgattt tgtcgccagt ctgaaacagc aggctgaatc 12000
catcgggaaa acacgcgccg aactgctgga aatgaaggcg gcgcaattgg gcgtgtcggc 12060
agaagcggca ccgttcatta atgccctgaa gcagcaggag caagcattaa agaaacagca 12120
aaacgctatg ggcctcgctg gcatttctgc cgggcaatat aaaatggcca tgcgccagtt 12180
accgatgcag atcacggata ttgtaacgtc ccttgcgtcc gggatgccaa tctggttggt 12240
tgctgtgcag cagggcgggc aaatcaagga tagcttcggc ggcttgagca atacgtttaa 12300
ggtattgttg agctacatca acccgctaac ggttggtgct gcgtcgttgg gtatcgccct 12360
ggcggcaatc gctaaagctg gttatgactc ctggagtgcg caacgagaac tggcgaatgc 12420
gctggtgctg actggtggct atgctgcgac caccaccggg cagatcacag ccttaaccga 12480
agagattaac aaaaactcat cggctacgat tggcagcatt caggaaatcg caacatcgct 12540
agctagttct ggtaagtaca caatcaacca gattaagcag atcacgaaga ctacggagga 12600
atggtcggcg cagacgggtg agagcgagaa gacgatcaca gggtattttg attcgatcgt 12660
aaaagaccct gttaaggggc ttgccgatct caatgagagc tttaattttc tgaaagaggg 12720
ccagctaacc tacatcgaat ctttgcgtaa aaccaaaggc gagacggcag cagtagatgc 12780
ggcgacaaaa ctgtttgctg aaacgatgga taagcgcctt aaagatatcg cggacagcgc 12840
aacgccgcta gaaacgatgt ggacggatat caaaaaatgg gccgctgact catggaaatg 12900
ggtaggcgat cacacggtcg gggcgctaaa ccttatcgtt gatacggttt ccgcgatcat 12960
taacacgatc aggaagctga ttgccgacgg cgacgccatg atcgcgcagt tcatcgttga 13020
cgcaggccgg aaactgcaaa aaattcccgg gatgggcgac tttggcaatg aatttctggc 13080
gcagcaggag cagctaatca aggactcgaa agccaaatcc gacacgcttg caaaaaccat 13140
tgcggaacag caggcaagga tcgcaaaagg cgagatgggt tacattgatg ccgcgaagga 13200
taaaaccctt tctggtgggc acagcagcaa aacgaaggat cgcgttaacc aggaagaaaa 13260
agatatcctg aaaaaccgca acgccaggaa gcagcaggta gacgcgggcg ttaaaattga 13320
tgagcagtac caggctgaac tgctatcgct acaggcgcag ctaaaggttt tgcagcagca 13380
taaagggctt gacgacaaga tcagccagca gcgcaaagac tacttcgaga cggtcgccaa 13440
atttcaggtg ctggaagagg caagccagaa gcgaaaactg aaccaaagcg aacagcagat 13500
gctggcgaac aaaaagaata tcctgtacat ggcggaacaa aaggccatcg tgggagatca 13560
gattgttcag cagcagcgac tgaacgcctt gcttgacaag tctaccaagt atcaaaacca 13620
gatggcggag aaaaccaaag cgctacagga taccgccgga atgggtagca aggagcagga 13680
gagataccgg gccaatgcgc agatggcggc tgactggaaa aacagcggcg gatctttaag 13740
tgaccctgga tttaaagcca tgcaggatgc aagtaataaa ttctacgcac agcaagatgc 13800
gcaaatgatg aactggaagg ccgggtttac tcacgcgtgg gctgacattg gcgacgaagt 13860
taatgacgta tacaccaaca tcggggatat tactaaaaac gcatttagcg gaatggcgag 13920
cgtgctgact gattttgtca tgactggtaa ggccagcttt agcgactttg caaagagcgt 13980
gatcactgac attaccaaca tgcttatcaa gatggcgctg tttaactcgc tttctgctgc 14040
gtttggtggt ggtggcggca cgttcagctt cgccaacatg ttcagcaagg gattttccgg 14100
cggtggttac actggcgacg gagggaagta tgagccaaaa ggcgttgttc acggcgggga 14160
attcgtattc accaaagagg cgacgcaaag gttaggcccg gaaaacctgt accgactcat 14220
gcgcggctat gcaagtggcg gcctggttgg cggtaacgca agttcagtat ctggagtaac 14280
caacggcggc aatgttgcgg cgtcggctgc gatggtgttc actatcggtg acattaacat 14340
tacgatgggc tccggtcagg atagtaaggg catggagcaa ggcgtgaggc agatcgtgaa 14400
cgatatgttc accgaggctt tgagccaaaa cgggcgcatt gcgaagttcg tcaatgagaa 14460
aacgaggagg tagtagtgga ttcctttaaa tggtgtacac aaattcaagg aggggcggca 14520
aaggtcgcca tctccaacaa cattcgatcg attgttttcg gaaacggcta catccaaacg 14580
gcatcgagcg gcattaacac aaagcgcagg acggttccga tcgtttatgg cggttcagat 14640
tgggaggagg tttataactt ctgtcaggat cacgtaacaa agccttttgt gtggaaggca 14700
ccggatggaa gaatgggcgt attcgtcgta actgccgact ccgttaacct tgcgccgcag 14760
ggtggcgggg tgtttgaggt aacggctgaa ttcgccgaac gcttcacttc agccggataa 14820
tcaaaaagcg ccctttacgg gtgctttttt ttggcctatg atctggagtc aattagagga 14880
gggcttacga tgacagccaa tgtttcaaaa gagtttgcga actgcttaca aaaacttttc 14940
cccggcgaga tcctaacgct gatcgatatc gacgccacaa agttcggcgg gcaggtctac 15000
cgattccata acgagaacgt cgcctataca accgaggagc ttttggcagc ggttaacggc 15060
gggacgctta agccaaagat gatcacgttt cgcggcgagc agtacggccc gcgcccgttc 15120
ggcctgggcg ggatcgcaat gtcgagcgat ggcacagtgg aaaagccaac gctgacggtt 15180
agcaatatcg atgcacaagc gagtgctctt attcgcgcct acaacggcct catgcaagcc 15240
aaagttacgg tatgggtttt ggtcaaggaa ttgctacaaa acgacggcag cgttaaagag 15300
ggcgatttta ggcgatttgt ctactacatc gagcgcccaa aacaggtcga cccgcaaaag 15360
gcaacgttcg aattaacatc cgtgtttgat atggatggat taatgatccc ggcacgccta 15420
acgcaaaccg tttgttattg ggcgcagcgt gggtggtaca agtctggcaa aggctgcgac 15480
tacaacgggc agaacggata cttcgacaag ttagggaaca gggttgacga tccgtcgcgg 15540
gatgtttgcg gtggcctggt gtcttcttgc aggcttcgtt ttggtaatga accgttgagt 15600
ttcggcggtt gtgcgacagc aactttgaag agtggtagct aatatgttga ctccgaaaat 15660
taaaatgcag atcatgcagc acgcgaagga agtctacccg cacgaatgcg ccgggctggt 15720
gacgcaaaaa tcacgcgtgc aaaaatatca ccgactcgac aacgtttcgc cagatcctga 15780
gaacgagtct atgccggacg aaacgcagta tgcgatggca gcaatggatg gggaaccgat 15840
cgccttcgtt cattcccaca ctggcgacgg tgcaaccaca attccgagcg ccactgattt 15900
atgcttctgt gatgagtctg gcttgtcgtg ggttatcgtc tccatcccgg aaggtgatat 15960
gcgaatcatt gagccgaagc gccgcccgct gattggtcgc ccctgggctt tgggtgctta 16020
tgattgctat ggccttatta tggattttca caagcgccac ggcgtcacgc taaaagatcg 16080
gcgtgttccg ttcgaatggt ggaagccgga atacaaagag aatctttacc aggactactg 16140
gcaagaggac gggttcattg aaaacactgg cgagcctgaa gttggcgata tgatcatttt 16200
tcagcttcag gcggagaagt ggaatcacgc ggggatttac gttggaaata acaacatcct 16260
tcatcacgca tatggcaagc tgtctcgccg ggatatctat tctggatggt acgagcagca 16320
caaggtttta atttgcaggc ataaggagct aaaacatggc atcacataca aagacgatta 16380
aactatctgg ttccctgggc cgtcggttcg gtgtcttcca caaacttgcg gttgattcag 16440
tcgctgagtg tatccgggcg ctgtcttacc aggttgaagg gtttaagccg ttcatgcaga 16500
gcaaagttgg ttcaaacatg cgcttcggta tcatcgcaga cggaaaacca atcagcacgg 16560
acgactttgc tactttcgcc gtggcaaggg agatcagaat catcccgatc ccaagagcca 16620
gaaagaacgg cgggttgttg caggtcgtta tcggagcggc gattatggtt gcggccttct 16680
ttactggcgg cggttcgctg gcggctatgg gcgctttttc gtcggcggct tttatggctg 16740
gcggctcaat ggttttgggt ggcgtaatgc agatgatcgc accgcagatg ggcggcaaca 16800
tgcgtgcaag cgaatcacct gagaacaagc catcttatgc gttcggaggg ccgattaaca 16860
ccacggcggc gggttatcca atccagttgc catacggtta cagattggct ggcggcgcgt 16920
tgttcggttc aggatcttac gcagaagaca acaactaatt aagccattcg ctttttagcc 16980
tgggggcgta gcctccgggc tttttgtcgt gtacaattgc gagactatta acaggaggct 17040
aaacgatgac taatatcaag gcccgcaagg gtggttcaag caagccacgt actcccgtag 17100
aaatgccaga caacctgatc tcaaaagata agatcaagtt gttgcttgcc gtttcggatg 17160
gcgaggtggt taacgacttc agcctgaagc agttgcattt tggaggcgtc ccggttcaga 17220
acgaggatgg aacattcaac tatgagggcg tgattgcaga gttccgcccc ggcacgcaaa 17280
cgcaggacta cattcagggc ttcagcgagt caagtgctga attccaggtt gctcgtgaag 17340
tcactcacaa cacgccttat acgcttaccg tatcgaacaa aaatctttct gctattcgct 17400
ttcgcctgtt atggccgcgc gtgctgactc aaaaagataa cggcgatatg gtcggatcgg 17460
ttgttgagta caagatcgag atggcggtag atggtgcaag ttatcagacc tacataactg 17520
gcaagattga cggtaagaac acgactggcg gttacgatcg gagcattcgc gttaacctgc 17580
cgcaaaactt cacgtcgcag gtgcttatcc gcgttagtcg agtaacgccg gacgctgacg 17640
gggtgaaagt tgtcgacgct ttccgggttg aatcctacgc tgaagttatt gatgcaaaat 17700
tccgctaccc gttaacggcc atgctttacg ttgagttcga tagcgatctg ttccagaacc 17760
agatccccac tatctcactc aaaaagaaat ggaagattat ccaggttccg agaaactacg 17820
atccgattaa tcgcacgtac tccggaacgt gggacggtgt tttcaagtgg gcgtggagca 17880
ataacccggc ctgggttctt tatgatctga tcatgaatca gcgctatggt ttagaccagc 17940
gtgagttagg aatcccggtt gataagtggt cgctgtatga ggtggcgcaa tattgtgatg 18000
aacttgttcc tgacaatcgc ggcgggatgg aaccgcgcta tttaatggat gtaattgttc 18060
agtcgcaggt tgaggcgttc cagttggtaa gggatatttg ttccgcattc cgtggaatga 18120
cgttctacaa cggtgaaagc ctatcgatca tcgtcgataa gccgcgcgat cctgtgtacc 18180
tgtttacggc tgataacgtc gttgatggcg ttttcgttcg gacgttccca agcgaaaaga 18240
cgatgtacac gtcgtgcaac gtcatgttcg acgacgaaga aaaccagtac gaacaggatg 18300
ttgaaccagt gttcaacccg gacgcagcaa tgcgatttgg ccacaacccg accagcatta 18360
cagcgatcgg atgcaccaga aggacggagg cgaaccgccg tgggcgttgg attctgcaaa 18420
cgaacctaag cgccacaacc gtttcgtttt ctactggcct ggaaggtatg attccttctt 18480
gcggcgatgt tatttacgtt gcagatccgc actggcaatc ggcctttaac ctggtgctat 18540
caggacgcgt tatggaagta tctggcgtgc aggtgttcct ggcctaccgc tgcgacgcga 18600
aggctggcga tactctgatc ctgaataccg acgacggcaa gcctgtgcgc cgcacaatcg 18660
ccagcgtttc ggctgacggt aaaaccctaa cgctaaacgt gggatttaat tttgacgttg 18720
cgccggacag tgtattcctg atcgagagcg atcagcttgc agcggaacag tatgtagtaa 18780
cccggattga aaagggtagt gatgacgacg aattcacctt tgccatcacg gctacgcagt 18840
acgatccgaa caagtatgac gcgatcgaca acggggtaat taccgatagc cgaccaactt 18900
cggttgtaga cccggattca ttgggcgctc cgaaagactt aacgattagc tcgttttctc 18960
gcattgttca gggaatgagc gtcgaaacga tggtgatcgg ctggtctgct gtacagtatg 19020
ccaaactgta cgaggtacaa tggcgcaagg atggaggtaa ctggaacaac gttccgcgca 19080
ctgcgacaac gcaggttgat atcgaaggca tctatgctgg cgagtaccag gcccgcgtta 19140
ggtgcattag cggcgggaat gtagcgtctc cgtggtcttc tttggctagt gcatcgctga 19200
ccgggaaagt cggagcgcca aaaggcccga ttaacctttt tgcgtctgac aacgagatct 19260
tcggcattcg cgttaagtgg gccatgccag aaggggcgga agacacggca tacattgagc 19320
tttaccagtc gcaaagcgga accgatcagg acgcaagcct gcttaccctg attccctacc 19380
cggcggctga atactggcat tcaattttgc ctgctggcta cgttaacttc tataaggctc 19440
gaagcgtaga ccgtatcggc aacgtttcag cgtggactga ttacgctcgc ggcatgtcgt 19500
ctactgacgt taacgccatc acggatgtaa tcttggatga gatcctcgac agcgacgcga 19560
tgaaagaact tcaggtgagt gcgcaggata gtgcggcaaa actcaatgaa tacgctaaca 19620
gcataatcca aaacgcatta gcgaatgatg gagatgttag aatcatgaga aaggagaatg 19680
gaaagaggaa agccgaaatt aaacatgcag aagttctcat agcaaatgag acggaagcca 19740
gggtgcagca ggttaatcag atctcggcag agttcaacga aaatctcaat gctggattaa 19800
ctcaagttaa cgaggcgctg gccaatgaaa ctgaggctag agttacttcg gaagaggcgc 19860
tttcagcaag gatcggagag aactccgcag cgctagatca gaaactcgac tcgtgggcag 19920
acgttaatgg cgttggttcc atgtatacga tgaagctggg cttgaagtac aacggccagg 19980
aatataattc cgggatggct ctacagctta ccgcgcaagg gagcaacgtt gtttcgcagg 20040
tgcttttcat tgctgataga tttgctatta tccgtaatgc agcatccggc gcgtacacgc 20100
taccgtttgt tgtgcagaat gaccaggtgt tcatgaataa cgcgctgatt caggatggct 20160
caatcaccaa cgcgaagatc ggtaacgtca ttcagtcaaa caactatgtt tcaggtagtc 20220
gaggatggtc aatagacaaa aacggaacct ctgagttaaa tgaggtaaca gtgaggggga 20280
caatttacgc tactgatggt tggtttaagg gtactgttta tgcagagcat attgaaggtg 20340
acgtgatgat cgcagaatca aacacgatac cattcaagaa atattcagat gttgatagcg 20400
gagattatga aatactgaaa attcatggag aaaactttga taggagtatc gatacgaacc 20460
tgattataca agtgaattct acattcagga atacttttag agtaattgtt cagactccag 20520
ggaaggggga aatagaattc tacagtatag acactggaaa tgacggaggt acaagagcat 20580
acgcgctacg tggcttcgat ttgccagcag ctggaagagg gcaagagaat aggatcatcg 20640
ttagggtaac tcaaagcggg aggggtagca gcataaaaac attcactcca agggttgaga 20700
gagataatgg aagcactgga caagaaatgt ctgatggagt catatataac actggtttta 20760
tcagagagag ggcatacata tcggcgtaca ggaaaggaaa taggattgta gtctaattac 20820
aaagcgccct atgggcgctt ttctttatca agtgatgttc ataaaaagaa aggggccgaa 20880
tggccccttt ttcttatgtt gtattgatat acttatattt catatcgcaa aaaggtggat 20940
gtttttttcg tcaactcctg ttgtttcggc ctgcgcaaac cgttgcaggc cgtagttggt 21000
atgaatagtt gcttattaca ttaattagga atcctattca aaaaacatca aggaagcaaa 21060
gtaacatcaa caaacctgcc aacaccatca acgttagtta gttgaactgt aacagttgat 21120
gcatctgcat tgacggaaac tgcaactgtt acaccagtag cacctgttac tccccacaat 21180
ccagtaggtg acgagaattc ctttccctgc agagtggata cagccgcagt cgcgctaatt 21240
cttcttataa gaatttcaac atcataagta ctgtaaactc cggacgatgt catagcttca 21300
atcagaattt ttctgcgctt tggtcttggt attgggtttg acgttgggtc agccaaagca 21360
gggtgtgaaa acgtaagtgt aagtgtagag ccagcccctg atgacaaaac ctgatttgtt 21420
tttgccccac caatattctg tcgtccagtg gtcatctggt acaacccatt tgttaccgta 21480
tcagaaacac caaggttgtt tctgaaaagt ttattgtcaa tggtaataga ttctcgcaat 21540
gcagatactg tgtagccagt taaatcgcac ccagtagctt gaaccgtcat atcagttgct 21600
gttcctccgg tggtaatgac acgttcaggt gtcaaggttc cagtaccgcc atcatcaacc 21660
ccttttgttg tcttaacatt agaaagcatc agatatgagc cacttccttc gatataaaag 21720
tgggtgttgt ataggacgcc ctcagcagac agtctagcag gccgctgtga gaataagtta 21780
ccgataatag ctccaccacc atttctaatg gcaaagtttg cagaaccgtt cctgtcaaca 21840
agctctccgg ttataatgtt gcctacagat ccataaatat tatacccaac gccatcgttc 21900
cactcgttac gcacgtttac gaatatgttg ttattcgccc cttcttgtaa atctacaccg 21960
tggctagatt gggtatttat tactacatca ataatctttg agtctataag gtttcttatg 22020
ccaatagcat tggcgcagat tgagcttcta cctacctgaa ttgatgtata tccataactg 22080
ccgacgccat acaggaaacg atacaggcca cagttataca ggatcccacc acgcaactga 22140
tttttgttgt tgatgaatgg tctggccttg tctcttccat caaatatgca gccgtaaatt 22200
cggaaaatag atccggggcc aaagatatag tcggccccca aagccttacg gattaccgta 22260
cccttcccta cgaaagaaga gtcaccagag actgtgtatg gcttgctgca ttcgccgtaa 22320
atcgtaaaac ctcctggcaa aattacagaa ccatttaagg tgtagatccc ttttacggta 22380
tgggggaacc tgacagattt gtaaccagca tcaatgatgg cttgcaatgc gtaatcaacg 22440
gcaacctctg ctcccggtgt attaagcatt gtattaatat cagcctttgt cattttgctg 22500
aatatgtttc cttctgctag ttgaattagc gtagcattac caatattaac cttgccacca 22560
ccatcaggct gcgccagcgc caccagcgtt gactgctcgc cagcgtcatt aatgatctgc 22620
tgaatttggt atcttgtagt ctccgctgca acctctgaat tttttgccga attttcagaa 22680
gattttgcgt tatcctctga agttttcgcc ttactttcag acgcagccgc agccgcagcg 22740
cttgattccg cagcttcgga gtctgcctta attttgttag caagattttg cagcgattcg 22800
aaatcgaact ccttaaagaa ctcgactgca tcagcaatta cggtttcctg tgactgatag 22860
tagcgcagag tttccgcaac atcttgcgcc aggccgtcaa cggttagcga gtcgcttaac 22920
aggatcgcgt aatcgctaga cgctacgaca gcgccttttg tggtgatggc tttaatttca 22980
gtatcgctaa ccaccttgtt tacgacggcc atttgaatcg gtgacgacaa aaacataatc 23040
gtcgcaccgg ggcgaatcag cgaaagcgat gattgccatt ttgtgccagt cccggtaacg 23100
gttccgtctg cggccatagc cgccttgcct tctctgtata gtgccatttt aaaagcctct 23160
tatgttggtt gagtaacgca gataatagca tcaatgaaca gataaaaaaa ggagcctttc 23220
ggctccttta gttgtcaaat tagaacggga tatcatcatc gaaatccatc cccgtattcc 23280
cgccgctgtt ttgcggtttt ggcggttgct gaactttagg ttgttgaggc tggccccacc 23340
cggattgctg attgccgccg ttttgcgaag gctcgcgctg gctgaattcg agttgtggca 23400
taatcatttc attgtggctg taaattgtgc cgttgtgctc gcggttcacg atctgaagcg 23460
tccggcaggt gacgctgatc accttatcca tttgcagcgc ttcgtcgtac cacttaatca 23520
tgttttcttt ggcaaagaaa acggcgcggt agttcgtgta aaccgtttcg tcctggccat 23580
cacgattgcg gatcttcatt cgctccgaca ggtctacggc gtacattttc cacggcccgt 23640
tattgttact gccttccttg acatacggtt cctttcggat aacacctgtt acaacatgca 23700
ttgtcgttcc tatggggcgg tttcccgccc ggttaaatta gttgaaagat gaaatatctt 23760
gcgcttctgg ttcaggcttt gattctacct tttccggctc ccgtttcgca acctcttgcg 23820
gtttaccagg gttaaagccg ttagccgggg tgactttcag ttctgcctgg cgtttggtga 23880
tatcgtcttc cgtcattttc cattccgcag gcgttaacgt ttgtttcgcc agcttataga 23940
tctcacgaag cgattcgaga tcttcgcacg cgtcaatgcg ttttttgaag tctttcggcg 24000
tcattttcgt aatttctgca tcatcatccg cctgcttgat gccgagagct gcggccagtg 24060
cataacggcg ggcgtaagat gttgttgagc cgtatgcttg ttcgacggtt ttactgatcg 24120
gcatattgta ctgaaacgcc atgaactcgc cgctttcgtg caaaaacata gtttcgaggt 24180
gcataacctt ttcggtgctg gtatccatca tggattgaat gaccataatt ttgttcttct 24240
ccagcgccgg ggaaatagcg tcgaggatat caccaagatt cgcgtaggtg ttcccaagat 24300
ggttgttctt cccgcttttc ttcgcggcca cgaagccaga ttttgccttg attaatgcgg 24360
ctgcgatggt ggtaaatttt tcagatgtac gcatgataaa gtttcctttt cctgattggt 24420
aatgcgcact atatcacaag tgcgcaccag tgtttagcta tttgtgccgt atacgtccgg 24480
gaacatgtat ttcacaaact gcggagtagg caaaacaact tccgccgcgt ttgactcata 24540
tgatggccat gaatcgtgct tcacgcactc agcatactga tgaatcacgc tttgatactg 24600
cttgcgaccg atctcgatct gctggctggt cagggtgaac gccagcggag caaacggtga 24660
ttttttctcc tgcgttagaa gtctgacaac tactgggcgt ttttcgttgt aagtcttcac 24720
gaacagatcg cgctgcaatg ccatcttgag atagtagccc aaattgaagg cgagtcgccc 24780
gaaatcgtcc ggcttggagg attgagtggt tttgtagtcg gtaatcacca cgacctcgaa 24840
aacctcatcc gggttgaacc cccactcctt gattagttct ggatcggaaa caacgtcaac 24900
atgatcgagt cgaaccttga ccttgacgcc gaagatctca ccgaagattg acaattcacg 24960
ctgtgcggta ggcgattcga tacatgcggc gtgtcgcggg ttggccaaca tcacgcttcg 25020
catttgaaca acggcatcga aatcaacatc cttaactagc ttgcgcccgg agttcatcgc 25080
ggcgctttcg tcgcaaagtt caatcgccca ccaaacattt acgtcaatcc cggcgcgata 25140
tgccatttcc agaagttccg ggtaatcctt gtttgacgtc ccaatcaggc cacacgcttt 25200
cagcttcgca gacagtgccg acttcgacgt aatcagatct ttaacctcgc ccggagaagt 25260
cgcccgcagg tactcgccat taaattttgc tgtctcaagc atacatgtat gcgaacaggt 25320
tccgaacgcc agcgcggctg tttcctcacg cactttgaat ttccagtgcg ccggggatgt 25380
tgcgtaaatc tcgccgaggc ttgagccgct aacgtactcc gcgcaccagg agttaggatc 25440
gtgatattgc tcgttagtca attcactgct ggtgtatgcc ctgaaaattg cttcagccat 25500
tgatattgct ccatttgtgg tttcgttgcg ttaagtatac gcatgacgat tcccggcgca 25560
agtcaaaaag tgctatccgc agttggttaa aaaacgagcg aaatttacgt aagatttagt 25620
aagatgcatc ttacgtggtt ttttcaatac atttcataaa gttaacgcaa atcggtaaga 25680
tggtaagatc cctataggta aatatccatg aaaaatctgg cgcgaaatca agcgaaaaaa 25740
gacacgtacc ccggagaaat cttaccaaga ttgatagata cagatagagt aataataata 25800
atattgttat ttatcatata cttactatct atatattgct gttaattggt taaattttgc 25860
gcgaaattta cgtaagattc atcttactaa atcttaccta aagtggttcg accagttgca 25920
agctactgaa tttaaggcat aaaaaaaggt aagactgatt ttctcaatct tacctaattt 25980
ctggtaaata tttaatcaga gaagagatga tgccttgtat aacttacgct caaagccgag 26040
cttaaagttg tcaccgttcg gaacgataac tttaagatcc cgatcgtcag cagccgccag 26100
catatcccga tctccacgac ggcaaactac ccgcatttct cgcttacctt ctccgccctt 26160
gcatttatac ctgtacgcca cgatctcgac attggaaggg atgatgcagg cccacacatc 26220
acacttgaat gaactcgcaa tattgaagtg catcgcctca atccaggagc gagcaaggta 26280
aatcggcccg ttaccgtcgt cgctctgatt ggtcactatc accgatccga aggtcagatc 26340
gccagctaac atcttctcgc gcccctcctc atcaatgaac agcatgttgc aatactcatc 26400
atccagccca tcttcatgca cgagttgcat cggtagcgcg tgaattagct cctgtctgcc 26460
gttctcgtga gtttttactc caacctgata tgatttgata tgctcatttt caatgccctc 26520
atagagcgtt acaggcgtgc tatcgacggc ctccgttctg tttaaaactg ccagcactct 26580
ttcatgatct gccatcttgc cgtagtcata cccgttatca cgagctacct gcttgtttct 26640
cttgaccacg tattcttgcg gaaccttgcc gagatagcgc ccaagaatgt tgatgcactc 26700
gctatacggc tggccgctta acttcattaa ccagccgatc cccttatcag caccgcagcc 26760
gccacagtat gcgccgccgt cgcctcgcgt ttctaatttg tcagtccagc ggaatctgtc 26820
tttgccgccg cagttcgggc agtcctgatg cttgccgttg aagtatcgag agtggatgcc 26880
gcaaatgttc tgcaacgctt cgcgccacat gccagccatg tacggcaaaa cctctttttc 26940
atcgtaaaaa tccacgtcgt cacctccaaa taaaaaacgc ctacacgaga atgataaccc 27000
ggcaggcgtt tagtgtttag acaaattgtg ctatcggact acgcggagca tttcccggcg 27060
gtcgcatcgg cgcgtaactg gcttgccgtt gctgtcaaac cttaaatctg gtcggcagaa 27120
tgaggcgcgg aaacctttgc aattgttccg gcggtagctc ttgtgtacga gataagcgcc 27180
atcggctgag atcatgccgc gcttacgcca ctgctgaaca acctggatgc taacccccaa 27240
ctcttttgcc gttccagcga tgccaccgaa ggcatcaata acaagctcca tccgcgcagt 27300
caacccggcg cgaacctcat ccttcagcac gtagtaacca gtcggtcgct tgcgtttctt 27360
cttatctttc ccgcgcgatg ttccgttatt gccgttcaag gttcgcttat ccacctttgc 27420
catttgttcc ataatttaac cctcatagca tttttttgtt aaacatgata aaatgttccc 27480
tgtattatac acgcaactat gcgaatgaca aattaggatt gcccatgctc acaattgaac 27540
aacaaattga agcctacgca gacaagattc cgctaataca aaagcggttc accgtcggaa 27600
atatcgttcc ttacccgtat caggcggttg cgtatattga gaccgcgaag cggatcgcaa 27660
aatatgaaca tcctttttac attaaggctt cggtttccgc cgggaaaacc atcatgatcg 27720
ccatgctcgc ggcgcagtgc aaggcaatga acttacccat gatggttctt gctcgccagg 27780
ccgagatcgt gaagcaggat tccgaggaga tcagtaacct cgatgttccc aactccgttt 27840
attgcgccgg gttaggcacg aaggccgcat acttcccgat tgtcgtcgga tctgaaggga 27900
cggtggttaa tggcctgttt aaaatgcttg gcgactacgt gccttcagtt ctggccattg 27960
acgaatgcca ccaggttgac tggcaagatc tggcggaagc gatcgccaac aatgaatcgt 28020
tcgagtacat gagtagaccg aaggataagc cgtatcgcgt gaacggggag ctggtcgatg 28080
ccgaccaccc atacgacgaa aaattcgacg acgtggaatt cggcggcggg cgcactcaat 28140
acaccatcgt aatcattgag ttaatgcgcc gttgccttca gaagacgggg cgcgagcttc 28200
gcattgttgg ctatacgggt tcggagtttc gcggcgtcat tccgatctta caggaagaca 28260
agagccagcc gggattctgg cgcgagcaga tcaccgacat taacacaaac tatcttgtcg 28320
agttcggatc ggttgttccc accattttcg gcgataccga ggcagatggg ttggggtatg 28380
atctttcaga attccacggc tccagtcagg acggcacgca ggattttagc gcggaagact 28440
tgcgcaagat ggagaagaaa atccatgaat ccggcgaaat gacgaagctg attatgcaaa 28500
aggtcgtgga gcgtgcgaaa accagaaacg gcgtcctgat aacttgcgct ggacagcggc 28560
attgcaagga ggcggcgagc tatctaccgc cggacgcaac atacgcgatc atcaccgaaa 28620
agacgaactc aaagaaacgc ggagaaattt tggataaggc gaatcgcggg gagattaaat 28680
acatcttcca ggtgatggcc ctaaccactg gcgttaacgt tccgttttgg gatttttcgg 28740
tgatattgcg caagataggg tcgcttacgt tgcttattca gcttttgggg cgtggaatgc 28800
gacttctaaa ggactggcaa aagcagccgc cttactcgtg ggtgaaagaa gatcaccttg 28860
tctgggattt tgccggaaca atggatgacc ttggccagtt gtatttcgat ccgattttgg 28920
agcaggcgca ataccaaaga cgtaagagca gcaagaacgg cccgaaaatt tgcccggtat 28980
gcaagggaga aaatagcgag tacgcccgcc gatgcatcca caaagacagc aacggtaatc 29040
gttgcgaata cttctggatc tcgcagcgct gcgaagacca gaaagacccg cgaacaggga 29100
agattaaagt aaaggggtgt tacgctgaaa acgatattgt tgctcgccag tgcagatgct 29160
gtggggtgca gcttaaagac cccaatgaca atctcaccgg gaagcactac acgcaaaatg 29220
actggtatga tgttgtcggg ttcgatatcg gattgactcg caatcagtcc gggatcatct 29280
tcaattacgt gctactgaac catgacggcg agcggttcac cgcaagggag aagttctttc 29340
cggaatcaga gaatcagatt tgcggcaagt tatggcggca aaaggcagtc ttccagcacg 29400
ttgacgacgc ggtaatgcgc ggcaagttgg gcgggatgaa aaatgcgcga aaaatccttg 29460
agagtgcgca ttacttccga gcgccgaagc gcgtaacgca tcgcgttaac ggtaagaagg 29520
aagatattat ttcacgcaaa gattttggag acgcagaatg atcacagata aaggtgatta 29580
tctcgaatac tacggcgggc ctgtaaaggc ttgcccgctt gagaaaatag accagatgaa 29640
cagcgtttcg tggcttcggc acgaataccc tgattatctg ttttggcata cggttaatga 29700
gggaagcaag cacaaagcga gcgcggttat cgatcaccaa atggggttgt tgaagggtgt 29760
tagcgacatt ctcatcctga ttggtttcgg tggcaaatac ccgttcgcag ccattgagct 29820
aaagcgccag ggtaaggcgc aggcttcgcc agtgagcaag gagcaaaggg aattccttgc 29880
tgccgtccgg cgtcgtggcg gattcgccgc cgtggcctat ggcttcgagc aattcaagat 29940
cgctttctgc gatgccatca aatagcactt tttgttaaaa ctgcccggcg aaagccgggt 30000
atcatttccc catcgaaact aagaatggag tgttgaaaat gaaaaagatg ctggctttag 30060
ttgttctgtc tcttggtctt attggttgca gcgaaaaacc gaaaacatat gattgcggtg 30120
gtgaagcgtt tgaggtaact agcaactata tgaaagttgt caagggtgag aactctggcg 30180
ttataattga tggcgctggc gaaaatcaat ataaactgct tactcctttt gggtacgctc 30240
attatgcagt caacaaaaac actattgatg ttagtgttgg cgcttttcac aataccttaa 30300
cctgcgaggt taaataataa tggcaaaaga tatcgcagac aaagaaaatc atgacgcgtt 30360
caccacattt gagcagattg agcgagaaac gttcatcggt aacgctcttg ccactggtgg 30420
acactatcag gccgtcaggc ccgacaagtt ttaccaggta acaggcaacc gatacgtcgg 30480
gagcaaaacg ccggatattg tgcgagataa gtgggcaacc gatcgcagcc tgatcgcata 30540
catggaagag cgttacggcc cttacgatct tgacgccgcc gcagaccaaa gcaacgcagt 30600
ttgcccgaag ttctacgacg aaaaaacgga ttgccttaaa cgctggtggg gtaaaaacaa 30660
gcacgtatgg ctgaatccgc cttactcgtt tccagatccg ttcgtgctca aagccattga 30720
gcaaatggag cacgataacc agatcgacat tctgctaccc ggcgacaatt ctacggcctg 30780
gttccgtgac gcgcagaaga tggccgccga aattatctgg attgttgccg atgttgaaga 30840
ggatgaggac gggaaccagt taagccgatc cggtcgcctc gcattcatca atggattaag 30900
cgggaagcca gtcgacaaca acaataaagg aagtgttatt ttcatcatgc gcaagctcaa 30960
gccgggagag gagcaaaaga cgctttacat tccggtaagc gagatttgcc cgtcattagc 31020
taaaaagcgt atgcgcaaac gtgggatctg aaaaatggaa caggtagaat ctttctccga 31080
gtatcttcgg attgtggttg aattgctgga caagtacggc ttcattggta cggatgagga 31140
aaagttagcc ttcgctgaca ccattgacgg aacctacatg gagttcatgg acaacggaac 31200
cccggtcgct gactggccag aaattcttga acgagaattg attgagttta aatcgcatga 31260
aggcgctgag tatttcgcaa aacagcacta attgctaaat aatgcccgcc gcgtgcgggt 31320
attattactc catcaaccaa tcaggagcaa acgccatgaa aaccaaaacc attgcagata 31380
ctatcaagat cgcaccagca aaagcgcaag ttgtatcgcg ccacctggtt aacatgtctc 31440
gcctgtgcat ggccgactac atggcgaacc cttcagagaa tggccttgat ggtgcaattg 31500
gcgagattta ttttcgcgcc gggtacggcc tggaaagtgt ggccatgtat gagcaaatgg 31560
ccgagggttt ttgcatttac ggtgacgaat gatgattgta gagactggtc gcgctgccgt 31620
atggcagcac gcaaaagagg ctggaataag tgatgatatc gtgaagatcg caaagtattt 31680
cgatatcaaa gatatatcaa ttatctttgg tgggaagctc acctatctac acgaacgccc 31740
tgtgaagcgc acgcgaatag cagtggcaac gcgagcggag gcggacgcgc tgaagatgtt 31800
catccacgag tcgaagcagc agaagaaata ttacaagtag cgggagtgtc gagaatgcga 31860
tatattgcga tcttctttac tgcgatcctg ttcacgatcg caatcattaa ctatgcaatt 31920
caatttggat aaattatgtc acctaaaatc acagacgaag aatttttagc cgcccgcgag 31980
gaaggcaaaa cctaccgcga gatcgcggaa gagttcggca tgaacattcg aagcgttgaa 32040
cgtcgcggag ttcgcctggc gcgacaagga cacctacacg gaaacgccca cgttgcgaag 32100
catatcccgg acggcttcgg cgtcaaaggc acgtcgacga tgattcgcgc ggacggctcc 32160
gaggtcgttc ggtgggttaa gtcggaagta gaccgcgatc gcatggttgc gcttatggag 32220
gcggcgcagg cagccttctg cgaagacctt ccgcgagccg aaccgcaacc gctggatgag 32280
tcgaagttct acattgaaga tcagcttgca ctgtacccga tcttcgacct gcatattggg 32340
gcaatggcgc ataagcatga atgcggcgag aactacgata ccagcacggc agagaaggtt 32400
ctaaaccgct tctttgatta ttccgtttcg gtggctccgc aatcacaaaa ggcggttttg 32460
ttggtcggtg gtgacttcct tcacagtgac ggactggacg cagtaacccc ggcaagcggt 32520
cacgttctcg atcaggacag ccgatacgca aaacttgttt atgtggccat tcgctcgctg 32580
cgtcgcgccg tgtcgctact gcttaacaat catgcggaag ttgaagtgca ggtgattgaa 32640
ggcaaccacg accaggccgg gatgatttgg ctacgcgcag cgttggcggc gttctatgag 32700
aatgaaccgc gcgttttcgt tgatgttagt ccggcgatcc tgcatcgcac cttgtggggc 32760
aaaaccatgc tgggctatac gcatggccac acgatgaaaa agccggaaac gcgccttgct 32820
gcgatggcta ccgacttccg taaggagttc ggccagtgcg actacattta cacgcattcc 32880
ggccactggc atcaccagac tgtaacggaa cactcgttag gcattgacga agtgcatggc 32940
cagttaggcg caaaagatgc ctacgccgca cgcggcggat ggcgttcata ccgccaggct 33000
gcggtgattc tgtacagcaa agaatatggc gaagtaggcc gctttatcta ccgtccgaac 33060
atgtaaccac aacggccccg cgaggggcca acaaggaaaa ccgatgaata gaaatatctg 33120
cattttcgat ctcgatggca cgctttccga cggaacccac cgcttgcacc tgctgccgaa 33180
aaaagatctc caccttacag aaagttggag cgaatttaat ggcgcgtcaa ttggcgacag 33240
cccaatccaa agcactattg acgtggcgaa tgcgctttat cgatccggaa tgaccgttat 33300
catcctgact ggccgatccg atgaggtgaa gaccgaaaca atgatttggc ttgaccgcta 33360
cggggtgaaa tatgacagcc taatcatgcg ccgcgccagc gataaccgta aagacacggt 33420
aatcaaggag gaggagttac gcaaaatcgg acttgatcgc attgttgcag cgtgggatga 33480
ttcccccaat gttattgcgc acttgcgcgg cctggggatc acgacttacc aggtctgcga 33540
ctacggcgac aatcttcacg atcatttaaa atcacacgga gtagacaaat gaaaaatgta 33600
attatcctca acggagcgcc gggcatcgga aaggacacta tcgcggaaat catctcgcgg 33660
aagtggcaat acaagaacct tagcttcaaa cagccgatgt ttgccattgc tcgtgctgtg 33720
ctgggatcgg ctgattttgc acgctttact gcccgatacc acgaccgcaa gcacaaagaa 33780
gtgaaatgcg attttttggg cgaccgttct ccgcgtgaat tcatgattca cattagcgaa 33840
aatttcgtca agccgaccct gggcaaaaat cagttcggca agttgctttg cgattcggcg 33900
ctaacttcgc cgtttaactg catcgtcagc gacggcggat tcgatgagga ggtggagcac 33960
gtcgcagcgc atgaggcgct taacgtgttt gtcgtccgcc ttcatcgtga cggcatgacc 34020
tttgagggtg atagccgaaa gcatattcga cgcccggatc ttatttgcga cacttaccac 34080
gaactcgatt ttgatatgac cactggcgag ccggaagacg acgcgcaaaa aatccttgat 34140
atggtgtcag atgttgcatt aaaattataa agttaatgcc tttattatca tcaccttaac 34200
tattgggaac cttgatgggt tccctttttt ttgttcttaa tttggcctaa tgcatatatc 34260
atcaccttac catttaacta acagaggttg catatcatgc gagaattcat caacgcggca 34320
accaatagca gcggtggcgt tgccctcgcg ggatctgcaa ccgggcaatt aatcattgct 34380
gccattggct tatttttcat ggttctattc ggctccttcg gcgcgtggtt gcgctggcga 34440
gattcaaagg cgcttcgtga agcgctggaa gccggggata tcaaaacggc ggtgaagatc 34500
aggagtaaat aacatgggga ttaaaacgcg ggttacattc gcggcggcgg tggcgatcgc 34560
ggtcgcgttc ctccccgaag tggaggacac gaaatacaat gtttatatgg atatcgctgg 34620
cgtcccgaca gtatgcgaag gcatcacagg cccggacgtt atcaaaggca aaacctatac 34680
ccggtcagag tgcgacgcgc ttttaaccaa gcatatccag gtggcgaagc gaaccgttga 34740
cagcaaaatc aaagtcgatg ttccggacac cttcagggcg tcgatgtaca gctttacgtt 34800
caacgctggc ggcggcgcat attctggcag caccatgctg aaattaacga accatggccg 34860
attgcgcgag gcgtgcgagc agctatatcg ctggacgtac taccgcaacc cgaagacggg 34920
taaaatggag aagtcaaaag gcttgtataa tcgccgggtt caggaatatc aactatgcat 34980
taaggatctg aaatgagcac attaaacttt caacgagcgc tggccatcgg ctttatcgtg 35040
tgggcggctg ccgtcgtttc cggttgcgcg tcaagcgtcc caatcctttc cgatctggtt 35100
ggtagcaagc cggatatgac ggcgcaagtc ggcgcggaga acgttaaaca ggcggttggc 35160
gtgaccaaca aaacggatac gtcgagcaag caggagacca cgttcaaaga gtcggcggta 35220
ggtaaggttg atacgtcgaa caagaaatcg gtgacgacct ccagcattca cgccaaccag 35280
attacggcgg acaagatcga gatccggaat gatgaaagcg gaagcctgat tccgtggctg 35340
attggtggtg ttggggtagt aatgctggcg atcggggtgt tcagcctttg gcgggaaaga 35400
aaaaacaaag gggcgtaatg cccctttttc tatatgtacc gcttgacgtg caatagcgct 35460
actccgtctt catcgttaag cccatgctca accgtgttgg ttgcggccat tccttgatag 35520
agcaagatga gcgcggcacg caagtaattt tctggtgtga tctgtttcac gcaaacaagc 35580
ctgtgaacct ccgttatcag atcttccacc tggtttcccg aaaagctgtt catcactgag 35640
ttggtcaagg tgcatcatct cccacatata tctgttatcc atcccatcga acgaccggaa 35700
atcaaaacct atttccctgt tatccggccc cgtacaccac acagcgccgt tttttccgtc 35760
aaggtatcca ttcgtatagc ttcgcgctaa aaactcctta gagaccattg aggcgaacat 35820
acgttgcgat acgtttgcgg cttttgccag gcgaggcgct tcgcggtgca tatagacgaa 35880
cttcgcaaaa tcctgccggg tgaattcccg gctagattcg cagaacttgt aaatgtcaag 35940
aatgaacatc aattacctca tgaacgacgg gttgataaag actttggaat ccatcacgca 36000
gataaagcct aattcctcca tcttcggcaa taagcgttcc ttaatctttt tgctaacccc 36060
ggcctggcct ttgaagatct tcaggttgcg gcaggcgtta taaatgccct ggacggtcat 36120
aacacctttc gcctgtttgc aacgactggc gataacgtca tacagcgctt taatttccgc 36180
tccctcacca gcaaagccgg aagagtcagc cgacgacaaa tacgttttgc tcaattcatg 36240
gaacatgatg atcgcttcgt caatggtcgc cgtgtcgatc ttcttcgagc gcttcccgcc 36300
tggttgccag ttccggatcg tgtgaatcac ggacgccaga cgcataacct gcttatcaaa 36360
cttacccatc gcgccgcgaa gcattgtatg cgaatactta ccgccgtcgc ctaactccgg 36420
ttctaattcc tggcgggcct tgttcagtct ccgcattgcc gcgtcagtaa cctgcaactt 36480
aacgtttgat tcgctcataa tgtcgtgaat cagccggaag taatccgcct tcagtgactg 36540
gtcgatcggc tcataggtcg aattcccgtt ttcgtcgatg aacacgcgct cacccaaacg 36600
ggtttgctca cgaaccaaaa ggaaacgctc cgagacaccg atcccgcgag aaccagcctg 36660
cataatggcg tcgatggttt catcctgtgc aattacgcaa atgcagccca aagccacgaa 36720
tgacatatta ttgctaacgt cggcacgagc gatcgatacg tggcctttat cccatgcttt 36780
gagcaccagt tcgctgttcg tcttcttgcc gccatcgttg ccatacgtga tccccaaaag 36840
gctgttaacc gccgtcgcct catcggaaat aacggcaaag tttccctggc ggttgttaat 36900
cttcgccagg ccttccgggg tggtatcgga taccgggaaa gttagatcac ataatttttc 36960
aagtttctct tcaagttcgt cacgatcttc gaaaagtttc accatatcag attgcgacaa 37020
ctcaccttta agcgcctgct tgttggcgga caacttcgcc atgattttct tacgctcttt 37080
cttgcgcgac tcgttaatgc gctcgacttc ggcgacgatc ggatcgatgg ccagcgagtt 37140
aatggcagac ttaccagcgg aaggcggctg cgatgtgata acgtaaagcg ttgtcggttg 37200
ctcgcttccg tggtactcga cccaaaatcg acccatcatc gcggcggaca cggttccgag 37260
aaagtgcatg taagcggatg attccgggaa ctgaacagaa cgtgctgcat tgagcgccag 37320
cttgccgacc acatcgtaat cattggcgat cgagattgtt gggtacttat ccgcgtttac 37380
gtcgatatct ttgggctttg gccagaatga aacggagtca cgatacccgt tcgcacgaat 37440
cgcaacacgc agaggactga taccctccct ttctgcgatg gcgataatat cttgcggtga 37500
tacgcggtca tttaaaaaca tgcccttgct cctgattggt taatcgttcg gctaatcata 37560
tgccgtatta aatccgagat ccagtgataa tctaaacacg cttgaaaaac gcgcttggat 37620
tatcggggcg catggccccg aacccgtcac aggtatttag cttcgaaagt cgttccgtcc 37680
gatacactga agccaacttc ctcgcggtac agcgtccagc ggcatccgtc acggtcgaag 37740
atgtatccag caaccgcacc gagcgcacga ccgctttcga cctggtagcg cttgccgacg 37800
ctgaaggatt ttttcatcgg gttgctatgg tcaaggccga cgcattttag cgttttggtt 37860
ttgagttcga tgaacgtcgc aaccgccgct ccgccgtcgc cagcaataaa caactcaccg 37920
ctaacgccta ccgacaaaat aacgcgcttc ttcttcagtt ccacgctgtc atataccatc 37980
attgacacgt tgccttcatc gtcgacgcgg gcggaataaa ggttgttttc atggatgtta 38040
acggcacggc ttgacgtgca tttaatcttg attgacttcg ccatttattt tctttctccc 38100
atctcgatat acaggttaac cagatccaga aagtcagctt tatttctgca actcaattta 38160
aagccaacgt gcaactcaat tttattttgc atcgcatcca gggttgcgcc gccattattc 38220
atcctgagaa cttcgcggca tacttctgcg agtttttgag atatcatttc ttcaccacct 38280
ttatttccat gccttcgcaa gcggccttga tatcattaat cagacgatta agccttaaat 38340
cagtcattac atggtgatga cgaacatcaa taatcccggc aactactaac agcatgacgg 38400
caaagataaa accaccaggc ccggacatac acaaaatagt taacatcatc aggataataa 38460
atttcatttc cgtttactcc gtttcgtttc gatgtggtaa tgctacccga ctttcgccgg 38520
gtagttttag caattcgtgc tattctcgat aattctcctg gaacacggcg cgggcaaaac 38580
ctcgcggagt aatggagcgt agcattttag tcctttctga ccttccgccc aaaaacttcc 38640
aggcccaaaa gaaattcacg ccttctactc catccggcgg cgggagccgt ttcggttcga 38700
caaaaccgtt tccgtgccag atgcacgttt tcttcgtgta gttgtcgcag tgtggcattt 38760
tgggatgcca gacaggctca tttggtgata catagccgcc gaagtcgcgc gggtgaaaat 38820
aaaagtctgg cttacgccat agtgtcgaca atttgccaac cgggttttcg accatccagg 38880
ggcaaccata ctcattgccc aattgctcga ccatcttcgc atcatcggcg gctgatagaa 38940
catcctgaat attccgggcg tgctttaccc cactatgcgc taacagcgtg caggacggga 39000
acgcgaaaat gaagtcagga tctggaattc cgagaatcga tcgcttcacg tcaaaatctt 39060
tgtcaatcca gatattaaca tactggatat ttgggtgaac catgcggata ttgtattcgc 39120
cgtggtcgcc ggagtctgca ttgaaacagt acaccttgca tcccttaatg gcccagggca 39180
ggcccatgat gccggagccg tcgaacatgc agtaaataac cttgtctttc atcatcacct 39240
caccagaaag ggacgcattc gctgcaacct tcgaagcagg agcagccgca cgtattctct 39300
tcttcatccg ccaggtcgtc gcacatcggc aggctttcgg cgttcatatc catatcacca 39360
agcgcatcag caagcgtcat ttcctgattg gcaaccatga tgcaggcttc acggttaagg 39420
ccagcgtcct gagccgcaag gaagcgactc cagtatgcat cgtcgtaatt atcagccatt 39480
actttttctc ccaatcatgc acgccagctt tcgataaaca tgctattcga ttccagcgtc 39540
agcgtgacgc cagccaggga ggactgaacg aggcgaccgc ccggaaccgt cccgatgatg 39600
taacggccct tcgacttcgt gatcgtgatc tgagtgaagc cggaggcgcg gttcatgcgg 39660
agaacgacgc ggccatgtgc gttaaggacg gcgagaatgg aatcagtctt cggatggttc 39720
attgtgtagc tccttcgctt gttggtgtgg ggatagtatg ccactatccc caaattctgt 39780
tttagcaatt cgtgctattc gggaacgcag gtcgtgtaat cctggcagat gtagattgtg 39840
cgctgaacac gcggcgcggc atcctccagg ctctgaatct cgatatcgtt gctgtcgacc 39900
cggcagcgtg ttgcaccgaa gatgaaatac cccgtctgct tgtcgttgat ttgccacgcc 39960
ttgctttcgg gaaccgtatt gacgacaaac ggcgtgtcgt cggctttgaa gtcttcaggg 40020
tgacgcccgt aatagtggat gattgcagca gttgccgccg ccttaaactt ttccgtcacg 40080
tcacgggcgg cgatctcacg cgagatctgg atatcaacat ccggcgcgcc gaccatgtag 40140
gaaaagctgt tcgaaatggt aactgttcgc attatatttt aacctcgttt tggtggatga 40200
agtcggcgac gtaatggcat tcgatcgggt tctcgatacc cgcgagcacc tcgcacgacc 40260
cgttattcag tagctggcaa tgctggcact ctgcaccgcc aacactggca atgacaatga 40320
cgatcccgtt gcatgaaccg ccgaacattc tgtgcgggaa ggggtaagca tcgcaggtgc 40380
aaaccacctc cccggcccgc cgtttcctca tacctggccc ttctcgatgc agaaaaccca 40440
atcacgagcg tcaaccgcaa agcgtgcaac gccgtcgagc accacgaaga cagtaaatcc 40500
attgacgccg cgccccttct cgatatcggt tgcggtacgg aacagatcgc ttgcgccata 40560
ttgaatttta tccccgacga ctacatccgc gaatgctttc atgttgattc tcctgactgg 40620
tttcgatggg ttaactatac caggcctcgc ggcccggcgt ttagcaattc gtgcttattt 40680
cactgctttc ttcgctgcct tgtacgcttc tttgaaggcc ggaacatctt cgaccggaat 40740
ccagaagccg gagccatagc catcatcata gcaagggcat tggtcgcagt agtcgcccca 40800
ggttttctct tcgccgccga cagagaagcc agtggccatt tcgcataatg cttcctcaac 40860
cgcttcgatc ttgtctgcgt cgctatccat gataacgaag ttccacttgc cgttatactc 40920
gctgcccagg ttgattgatt cgcgttgaag tttcatagct ttagcccctt tggctcaatt 40980
cgtttcgttg aagtcatcat agcaaagtcg atcgggtggg ttttgacaaa aaatgctatt 41040
cctgcgattg ctgattattc caccgattag gaatagtgga atagatggtg attgcacctg 41100
attccccggt gattagtttt gacgaatcat gattgctttt gattgtttca ttctgcgcaa 41160
cgattttggg agtgtttcag gatttgcaac gaattgcata aatgtatgca ccatgcacgc 41220
aatttgccga tcgaagccag caaaacgttg ggtgtgtttc gccaacattt acgtcgtgtg 41280
ggaagttggg gaagattgaa ccttacctaa aatttgcgca aaaaatgacc aatataagcc 41340
catatataga tagtaagtat atgataaata ataatattat tattattact ctttatcttt 41400
atatatgttt ggtaagattt ctccgggggg gggggggtta tggattcctc ttgcgcaagc 41460
cataaatttt tagatgttcc gccgcgaaaa atgtaaatag gtatatatag tggatgacag 41520
caaatcttac caaacacgcc gaaacccttt gacggcaagg cctgcggcct gtattatcat 41580
cttccgaaac attacaaaac gccgtagatg tgggggaaac acaaatgcaa tataaaggct 41640
atgagttcga agaagtcgac gtgtccgagt ggatgcggta cgacgcagat aagcaggccg 41700
cttaccatga atggctgcaa tccatcacgt ttggtgacac caccaggcca gcaggacgga 41760
gccaggaagc ctcgaagccg gaaggtacat tcactggcaa ccgctcttcc gagatccggc 41820
gcatgttcct ggacggcggg cggcgtatcc agatgacggc ggaggagttc gcggagaagt 41880
tcggcgttca tccggtcgac aaccatttcc gcaggccgtt gctgaaactt ctggagccgg 41940
gcgaggtgtt gcgagtgagc ctgggggcgg ggttaatctc cgtctttacc gagttcgacg 42000
caatgactgg ctcaaacctg gaggctgagg catacaggcg gcagggcgct atctctgagc 42060
gggaaagaat cctttccatc atcaacgaag taatgcctga aggcatcatg ctaaaaggct 42120
ataagctggc actaatcgcc aaaatcacag gagaataacc atgaaactat atccagacca 42180
aaacgacccc gccgcaccga tgcgccttgg cgacacgcca atcgacgagc aggaggttat 42240
cgaagccatt gccactgcat cggacgacga aaatccaacc gccctggaat gcgttacagc 42300
cgacttgctg ttaatgtcat ttcttccgga atctgaatct tacattcagc acatcaccga 42360
gacgggtcag gcggtgacga agccggaatt catcggtgcg tcgaacgtcc tggcgatgat 42420
ggccgatgag gccgacaaca tcattagcct ggattatcca caatgagcgc ctttatcctg 42480
ttttccatcg gtgttaattg cggattgctc ctggcggcat ttatcatcca cctgagcaac 42540
aagtagcact ttttgttaaa acccggtcag agcgatcggg tattatctct tcatcgacaa 42600
cgaaccgagg aaagcgaaat gattactctg attacctggg aacacgaaaa cagcaagccg 42660
gaagtacgcg agttcgagac cgtggcggcg tgctacaacc tggcggcaaa cggtggcttt 42720
tacaaggcgc agattgttaa cgagttcggg gttgtcgact atgaatttta aggcgagcga 42780
tgtaaaacct ggcgcggtct acggatcgcg cttcaacgac cgcctttggc gctgggatgg 42840
cgaaactatg tggacgaaag gcgaattcga cgtgatttgg catgaatgcg gctggcccca 42900
tccgacgatg gagcgccggg atatcgctta ctatctttca atcggttata tgcatgaggt 42960
ggtcgatagc actttttgct aaaacaccga tggcgagatc cggtattatt tattcatcga 43020
caacgaaccg aggtgatgaa gatggataag gttatcaaga tgagcgaagt taagccgggc 43080
atgatggtta agtttgctgg caagtttcgt ctggttctcg cagcagatcg caaggcgaat 43140
attctcacta ttcgcgttaa tggaaaggcc caactattcg cgccgcagtc cgacattgaa 43200
gttgaagttc gtatcaagta agcaacctgg agatgaagat gagcaagaag aatccatatt 43260
caaccagcaa ttaccataac gaacacgtca tgctctggca ctggcagaaa ctcaaagagc 43320
gccaccttgc acgcaagggc aaataatcgc aatagcgccg cttcggtggc gctgtattcc 43380
aaccaccagc acggcaaacg ccatgaggag ttgtcatgcc acgattttca gcaacaacca 43440
aacttcgcac cttcgccggg atgctgatcc catactcatc aaccaaggcc gtccagggca 43500
gcgagcacgg cgtatatttc cactggttag gtaagtggag attcaccgtc attcgcggct 43560
tttacgtgac ctgcgaccgc gtggatatcg aagaccattc cggcggaaac caggttcacg 43620
agttcaaaaa gcacgaatag cactttttgc taaaactcga tcggggcaat gcgttattat 43680
taccccatcg aaacagagga gcaaaaacat gattggcaac cacaacaacg aactgaacgc 43740
agcagcccac cgtcgcgccg tcgagcagaa cttcaacgcg cttaaagtag cctgcgatga 43800
gatgaaagca atgcttgaac tgccgtcatg ggatccccaa ctagaagact actatgacgg 43860
cctccgcgtc aagcgtgacg atatcatcaa ccgcctgcgc ctggccggaa tgttcctgta 43920
aggagaacgc aatgattcac gaagttaagt ccggcgagaa agtagtggcc accatcacgc 43980
agcgccacgt tgtagccttc cagattaaca tccctggcac gttcgatatg cagtcgcttg 44040
acgtgccagt ttgggccaac acggttgcca ttgatgccga tggctcaatt tgggcctatg 44100
agtcgaccgt tgaaaatgtt cgcctactga gctactcgcc tgattcatgg gtgtataagg 44160
ggccgggcgg aaacagaatg gttcaggtgg gcgaaatggc tcgcttccct gattgggcaa 44220
aatcgaagat tgaccttcgc ggcctgaaat agcacgcatt gctaaacgcc cggtcagagg 44280
tcgggcataa ttaccacgaa gtcgattaac gacccaatta tgcaaggaga caggataatg 44340
cagcaattca aatctcgcgg cagggtatac agcctgccag ataccgcaac tcatgccgcg 44400
cccggcgtat gccacggcct atacttcatg gatggcggca agtggttctt cattggcgat 44460
atggtaacgg atggcgtgca gccgctcctt tgcggcaaac atcgcggatt ctatgatcac 44520
gatatcgttg agttggagaa gaagcgcgat ccgttcggtt tttggaacaa aatcaaaggg 44580
gcgatcttta aatgaaccaa ttcgaattcc tggcgcgggc gcgccgtgcg cttgcaatgt 44640
acgccttcag caagaacatg aagattatca gtgttgaggt tgagttcaac aatatcagca 44700
tttccggcta tgtgggcggc tcattacatg taacaaaatt cagctttccg tcgctggcca 44760
gcctggagcg tgaggcgttc ggtttctgaa tagcactttt tgctaaacgc ccgaccagaa 44820
gatcgggcat aatggccaca taccaaacaa cggagattta ccatgattcg cactaccagg 44880
ccgctaccca ataaccttta cgccacgatt gggattcgcg cggtgagact gattcacgtc 44940
aacgaagttg gcaacgtggc tattttgtgg tttaagcgcc actcaagcgg ggttggcctg 45000
aacattgaaa gattcgggcg ctttaacggc aactcccaaa cccattctca ttgggcggat 45060
ggaacttacc actggacgca cgttaagcac gtcaagaact tcgacgccaa atcactgagc 45120
gacgacgacg cctggaagat ggtaacagat ctcttcgggt atgatgcctg catggaggta 45180
gttggatgat acgtttatca gacttcgatc gccactgctt aaccgggcaa tttggcgaaa 45240
agccagtgtt gtgcaagatc tcgaaggtta agggcgagcc gtttgagcaa atcaggacgc 45300
ttataagttt tgccagacga aaccgccttt acatccaggc acgcggagga ctgttcgcaa 45360
agcaggttca ttttgcgtat ggaacagggt tctacactgg cggagacggg gttaacccat 45420
caccgaaaag ggttaggctg cgcgagataa tcgaacgcgg ttacgtttgg actaacggga 45480
tctactaatg gaaacggtta aagtaatttg cacgcacgtc ggctattcgc cgctatcaac 45540
ctatttcacc cctggccgcg agtatgacgc caggttcggc cctggccttg atgaagtgtg 45600
gattttgcag gacgaccagg gaaccacgga ggacgaagaa ttctggccag cctgccgtat 45660
gccggacgaa aagatcgcaa tgtacgccaa taggccatac gaaaacaacg ctttatttga 45720
ggtgaaagta tgaaagtaag caaaaagaaa ctcgcgctgt attgggcctt cttcctggtc 45780
ggcctggtac tgcatgacca cgtaggctgg caggcgatcg gcatgatttg catcatcatt 45840
ggggtatgcc gcctgtctga gattaacggg ttccgccgtg ggtatcgggc agcgttcagc 45900
ggcgaaaaag aatagcactt tttgttaaaa ctgccgtaac gtcatttgat aaagtggcgt 45960
tactgaagtg aacaatccaa tcaggagcaa cgcaatgaac gacaatttaa tcgcactgct 46020
ggaagaactg aattcctacg gctgcgcata catcgagctt tcgagcgagc aggttattga 46080
agtcgcaatc gacgatgacg ccgggttcac cagattaagg gtggcggcag gggccgaaga 46140
ggaattcgag gactgcggca tccgtgaagt aaccaacctt ctcgaatccc accaggtgaa 46200
ggagatccga taatgaaaac cgtcaaactt atggttgtca atgcggcccg cgaacgtgaa 46260
cgccacctta tcggctgcgt attcgatgcc gtcgaggtgc acgaactagg cacagtgcac 46320
gtaatccagg gcggatacct gaacgaattc atcgtcaacc tggacaccga cgaagacttg 46380
accatccgtg ttggtggcga gccggaaaac gcgcttttcc acctgcaagt atcgaaggcg 46440
tcgcggcgca agttgatggt tcgcgcactg aagcgcatag cgaagcagcg cggatccgac 46500
gcatacatcc gcaagttcgg cacgttcgct ggcgtaggcg attgggcgcg gtcatgggcc 46560
gacttttact tcaacaaatc cggcgaattc tgtgccgcgc atacggactg gctgcatgga 46620
accgaacacg atcccgttaa cggccacgag tggtacgaat tcattgaggc ggaattagac 46680
gccctttgat agcacgaatt gctaaaacga acgtccgggg atggtgcata ctatccccac 46740
actaacaaga gatggagcct aaccatgaac accacacaaa tcatacacct gatcccgtcc 46800
gacaacttcg ccaaagacga cgacccgctg ttagccgccg tcattcgcgc cgggatcgat 46860
tgggcgctga gtgatgcatc gtgcgaatca ttcatgcgtg aaccattctc acgaatcgcg 46920
gaacttgaaa agcaggttgc tttcctgact ggtgaacttc aggcggcgca caccaagatc 46980
gagaaggttc gggaaatcat gcgcagcgaa cctggctatg acgttgatca cgcgcgagtt 47040
ttgcgcatga tggccgacgc attcgcaaaa ctccagcaat agcacgaacc acaaacacta 47100
accggggttg ctgcatggct accccaccaa agacaaggaa aaacatcatg atttacgcaa 47160
ttattgccct ggctgttctt gttctggccc tgtatgcgtg tggctgttta ctcatgcgag 47220
ccttcctgaa atctgctgat tccaatgaca aagacgatct ttatcctgtg ttgtgctggc 47280
cctggattac actttccgcc gttggcagcg tgatcatttc ccgcaatttt aagtggtgag 47340
atggcgcttg ggtgccaaaa tggatagcct gccaaatagc acgaattgct aaaacttacg 47400
caaggccatt tgatatagtg gccttattga agcgaaccaa tcaggagcta aacaaatgag 47460
cattccgtat ccgaccaaag acaaatcatt cctgggcctg gccgttgttg agattatcaa 47520
ccctgacgtt accacctccc aggatgagcc tacccttgca cagtgggccg gattcaaagc 47580
aggtgatcga ctgaccgtcg aatgctatag cgacggcgag aagtgggcgc aggtagggcg 47640
cgactaccac accgaagaat tcggccatgt tctccctacc gactggttca aacttaacga 47700
aggcgaatat aaggtgatcg aagaatgaac aacaaaattt gggtgctgac ctacactatc 47760
ggaactaacg aggggcgcaa agcgccgcct cacctgcgac acgaaagcac aggcggaaat 47820
gcagcaggcc gtgctgggcg gcgaggtggc cgaatacatc cgccagccag aatccttcca 47880
tgtcaattgg cctgagaaga tggatgttga cgccgtgctg catgaaatgc gccaggtgca 47940
aaacgaccag gcggcgtgga aagacctgta cctttgcggc gacgctgagt cagtactgga 48000
tcacttccgc tttgtccggc aggcgcacgc ggaatggtca gatcgccagt ttggcaacgt 48060
tgggccagtc gggccactca aacacctggc gaaagaggcg atcgaagccg ccgaagcgcc 48120
ggatgacatt agcgagttcg ccgatatcct gatgttggta tgggacgcaa cgcgccgcgc 48180
cggattcagt gatgagcagt tggttgaagc ggtggcggaa aagctggaac ggaacaagcg 48240
ccgggcgtgg ggcgaggtca aagatggcga accttgccac cacgtaaaaa attaacgaaa 48300
tcgaatgccg tttaaagcgc ctgtaagcca ttctggcggg caaatttaag gagtgcgatt 48360
gtatgcctga atactcaaaa gtcgaagata tgccgattgg cgcaacgatt actggtatct 48420
gtatgagtga gtctgtcgac gcaatcaagc cgctggcgtt cccggttaca caggtcgaaa 48480
ccgatcaaaa aaggtttcat cttcatctac aaaaatttca acgcccctct gcgtgtcgag 48540
atcttcattg ctcgcggaac ctgggtagaa tgggatgggg cttaa 48585

Claims (10)

1.一株裂解性肺炎克雷伯菌噬菌体RDP-KP-20005，其特征在于，其宿主为肺炎克雷伯菌BK-20005，透过电镜观察，该噬菌体头部为廿面体，长度为92.281nm，且立体对称，短尾，尾部长度为107.442nm，为有尾噬菌体目短尾噬菌体科，基因全长48585bp，不具有毒力基因和溶原性基因，保藏编号CGMCC No.21408。
2.如权利要求1所述的裂解性肺炎克雷伯菌噬菌体RDP-KP-20005，其特征在于，该噬菌体在pH在6-9范围内能维持较好的活性，其效价在5.9×106pfu/mL之上，在pH为7时，效价最高，适宜生长在中性环境中；在pH＝12以及pH＝2时，仍有噬菌体存活，该噬菌体的酸碱耐受范围宽。
3.如权利要求1所述的裂解性肺炎克雷伯菌噬菌体RDP-KP-20005，其特征在于，该噬菌体的最适温度为37℃，效价最高达到1010pfu/mL，在70℃的条件下作用60min，效价维持在105pfu/mL，依然具有生物活性，对温度有较好的耐受性。
4.如权利要求1所述的裂解性肺炎克雷伯菌噬菌体RDP-KP-20005在制备预防和或治疗禽源肺炎克雷伯菌引起的疾病的药物中的用途。
5.如权利要求1所述的裂解性肺炎克雷伯菌噬菌体RDP-KP-20005在制备预防和或治疗禽源肺炎克雷伯菌引起的疾病的饲料中的用途。
6.一种用于防治肺炎克雷伯菌的杀菌组合物，其特征在于，包括有效量的如权利要求1所述的肺炎克雷伯菌噬菌体RDP-KP-20005。
7.如权利要求7所述的用于防治肺炎克雷伯菌的杀菌组合物，其特征在于，还包括用于抗肺炎克雷伯菌的抗生素。
8.一种预防和或治疗肺炎克雷伯菌引起的疾病的添加剂，其有效成分包括权利要求1所述的肺炎克雷伯菌噬菌体RDP-KP-20005。
9.一种清洁剂，其特征在于，包括如权利要求1所述的肺炎克雷伯菌噬菌体RDP-KP-20005。
10.一种消毒剂，其特征在于，包括如权利要求1所述的肺炎克雷伯菌噬菌体RDP-KP-20005。
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