质粒名称: | pCAP03-acc(3)IV |
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出品公司: | 爱迪基因 |
目录编号: | 69862 |
存储实验室: | Bradley Moore |
载体骨架基本信息 |
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载体名称: | pCAP01 |
载体出品公司: | -- |
载体抗性: | 卡那霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | 在粟酒裂殖酵母 ADH1 基因 (pADH1) 的强启动子下,将 URA3 基因导入 pCAP01,将其导入 pCAP01 的 SpeI 和 KpnI 限制性位点,作为计数器选择标记,以产生 pCAP03。pADH1 可以容忍在 TATA 框和转录起始位点之间插入多达 130 bp 的片段。这种方法允许使用较短的捕获臂,并在 5-氟乳清酸 (5-FOA) 存在下方便地选择非同源末端连接。为了易于线性化 pCAP03,从 pIJ773 扩增 1369 bp 安普霉素抗性盒 (acc(3)IV) 并将其克隆到 pCAP03 中以获得 pCAP03-acc(3)IV。 |
载体类型: | 细菌表达、酵母表达 |
筛选标记: | TRP1、URA3;安普霉素 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | -- |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | - |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pCAP03-acc(3)IV was a gift from Bradley Moore (Addgene plasmid # 69862 ; http://n2t.net/addgene:69862 ; RRID:Addgene_69862) |
参考文献部分: | Identification of Thiotetronic Acid Antibiotic Biosynthetic Pathways by Target-directed Genome Mining. Tang X, Li J, Millan-Aguinaga N, Zhang JJ, O'Neill EC, Ugalde JA, Jensen PR, Mantovani SM, Moore BS. ACS Chem Biol. 2015 Oct 21. 10.1021/acschembio.5b00658 PubMed 26458099 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCAP03-acc(3)IV 11875 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pCAP03-acc(3)IV is a upgraded version of the pCAP01(Plasmid #59981). ACCESSION . VERSION . KEYWORDS pCAP03-acc(3)IV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11875) AUTHORS Tang X, Li J, Millan-Aguinaga N, Zhang JJ, O'Neill EC, Ugalde JA, Jensen PR, Mantovani SM, Moore BS TITLE Identification of Thiotetronic Acid Antibiotic Biosynthetic Pathways by Target-directed Genome Mining. JOURNAL ACS Chem Biol. 2015 Oct 21. PUBMED 26458099 REFERENCE 2 (bases 1 to 11875) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11875 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 733..766 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" oriT 773..882 /note="incP origin of transfer" CDS 1198..1998 /codon_start=1 /gene="aac(3)IV" /product="aminoglycoside 3-N-acetyltransferase type IV " /label=ApmR /note="confers resistance to apramycin (Paget and Davies, 1996)" /translation="MSLAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG" protein_bind 2008..2054 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2077..2880 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" misc_feature 2958..3461 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind 3502..3520 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3558..3580) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3680..3699 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 3720..4000 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 4001..4675 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" primer_bind complement(4915..4936) /label=F1ori-F /note="F1 origin, forward primer" rep_origin 5183..5771 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5672..5691 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5925..5942 /label=L4440 /note="L4440 vector, forward primer" polyA_signal 6156..6290 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6180..6199) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6234..6253 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(6715..6735) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" intron 6865..6930 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS complement(7350..8144) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7461..7480) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 8071..8090 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(8928..9299) /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" oriT 9332..9441 /note="incP origin of transfer" CDS complement(9679..9690) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind 9761..9860 /label=phage phi-C31 attP /bound_moiety="phage phi-C31 integrase" /note="attachment site of phage phi-C31" CDS 9870..11687 /codon_start=1 /product="integrase from phage phi-C31" /label=phage phi-C31 integrase /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL RAPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI EKRASITWAKPPTDDDEDDAQDGTEDVAA" ORIGIN 1 cctacaacaa ctaagaaaat ggctatcatg cggaagctgg tgagaagaac agcatcggga 61 caagggaagg aagaacaaag acaaagaaaa caaaagaaag caattgaaaa caaaacaaaa 121 caattttcat tccttctctt atcattcctt ttcttttctt ttctctcatt caacgcactc 181 catcgtatcc gtattcctct tattttttct ctttctctat atccatttct ttctctctag 241 gtgtgtcctc tctctctctt caatttctct actccgcatt ccaacgcatc cttcccccaa 301 cctcccattt cctccttacg gcccgatagc gatcgtcttt ccctcgctat cactcgctac 361 cggcccctcc tctgcaccgt aacctcctac gtatttacca tatcataaag ttttttccga 421 cgcttatcgc tgaccccctg ccgccctcct attggcttcc ggattatctt cttgtccata 481 aggtgatcca tgcttcctga agattcccga aatgtgtcca ctttggcggg gaatcattcc 541 atccacttct ttctctctcg ctttcctcat tcggcgctcc ccttccgcgt ctcattggtc 601 ttccgctccg tttttgcttt gccgatgtta cttggggaga ggtgcgataa tcctttcgca 661 aaaactcggt ttgacgcctc ccatggtata aatagtggct cgagattccg gggatccgtc 721 gacctgcagt tcgaagttcc tattctctag aaagtatagg aacttcgaag ttcccgccag 781 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag 841 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata 901 caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat 961 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaatgca 1021 gctcacggta actgatgccg tatttgcagt accagcgtac ggcccacaga atgatgtcac 1081 gctgaaaatg ccggcctttg aatgggttca tgtgcagctc cataagcaaa aggggatgat 1141 aagtttatca ccaccgacta tttgcaacag tgccgttgat cgtgctatga tcgactgatg 1201 tcattagcgg tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt 1261 cagcttctca accttggggt tacccccggc ggtgtgctgc tggtccacag ctccttccgt 1321 agcgtccggc ccctcgaaga tgggccactt ggactgatcg aggccctgcg tgctgcgctg 1381 ggtccgggag ggacgctcgt catgccctcg tggtcaggtc tggacgacga gccgttcgat 1441 cctgccacgt cgcccgttac accggacctt ggagttgtct ctgacacatt ctggcgcctg 1501 ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg cagcggggcc acaggcagag 1561 cagatcatct ctgatccatt gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt 1621 gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc caacacgacg 1681 ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt 1741 cttcaggatg gcaagttggt acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc 1801 tttgccttgg cggacaggtg gctcaaggag aagagccttc agaaggaagg tccagtcggt 1861 catgcctttg ctcggttgat ccgctcccgc gacattgtgg cgacagccct gggtcaactg 1921 ggccgagatc cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc 1981 gctcgccagt cgattggctg agctcataag ttcctattcc gaagttccta ttctctagaa 2041 agtataggaa cttcgaagca gctccagcct acacatatgt cgaaagctac atataaggaa 2101 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag 2161 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt 2221 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt 2281 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc 2341 ttcgaagaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt 2401 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt 2461 attgttagcg gtttgaagca ggcggcggaa gaagtaacaa aggaacctag aggccttttg 2521 atgttagcag aattgtcatg caagggctcc ctagctactg gagaatatac taagggtact 2581 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg 2641 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac 2701 aagggagacg cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct 2761 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt 2821 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa 2881 ggtacctgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 2941 ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat 3001 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt 3061 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc 3121 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca 3181 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct 3241 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc 3301 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa 3361 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca 3421 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccattatt 3481 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc 3541 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 3601 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 3661 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagatca 3721 cgacattact atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg 3781 tactttgcag ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc 3841 ttgtcacctt acgtacaatc ttgatccgga gcttttcttt ttttgccgat taagaattaa 3901 ttcggtcgaa aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac 3961 gtgagtatac gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac 4021 aggtagttct ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg 4081 tgctctagat tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac 4141 aattgacccg gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc 4201 aggcactccg aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc 4261 tctggtcaat gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga 4321 ataccaagag ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa 4381 catactactc agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga 4441 agcaggtggg acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca 4501 agagagcccc gaaagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg 4561 tgatgcgctt agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa 4621 tggtgtaaaa gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat 4681 tactgagtag tatttattta agtattgttt gtgcacttgc cgatctatgc ggtgtgaaat 4741 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg 4801 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc 4861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt 4921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc 4981 tatcagggcg atggcccact acgtgaacct aactgtcaga ccaagtttac tcatatatac 5041 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5101 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5161 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5221 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5281 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5341 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 5401 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 5461 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 5521 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 5581 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 5641 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 5701 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 5761 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 5821 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 5881 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 5941 aggaagcgga agagcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag 6001 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc 6061 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc 6121 ctcaacgaca ggagcacgat catgcgcacc cgtcagatcc agacatgata agatacattg 6181 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 6241 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca 6301 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt 6361 aaaacctcta caaatgtggt atggctgatt atgatctcta gtcaaggcac tatacatcaa 6421 atattcctta ttaacccctt tacaaattaa aaagctaaag gtacacaatt tttgagcata 6481 gttattaata gcagacactc tatgcctgtg tggagtaaga aaaaacagta tgttatgatt 6541 ataactgtta tgcctactta taaaggttac agaatatttt tccataattt tcttgtatag 6601 cagtgcagct ttttcctttg tggtgtaaat agcaaagcaa gcaagagttc tattactaaa 6661 cacagcatga ctcaaaaaac ttagcaattc tgaaggaaag tccttggggt cttctacctt 6721 tctcttcttt tttggaggag tagaatgttg agagtcagca gtagcctcat catcactaga 6781 tggcatttct tctgagcaaa acaggttttc ctcattaaag gcattccacc actgctccca 6841 ttcatcagtt ccataggttg gaatctaaaa tacacaaaca attagaatca gtagtttaac 6901 acattataca cttaaaaatt ttatatttac cttagagctt taaatctctg taggtagttt 6961 gtccaattat gtcacaccac agaagtaagg ttccttcaca aagatccgga ccaaagcggc 7021 catcgtgcct ccccactcct gcagttcggg ggcatggatg cgcggatagc cgctgctggt 7081 ttcctggatg ccgacggatt tgcactgccg gtagaactcc gcgaggtcgt ccagcctcag 7141 gcagcagctg aaccaactcg cgaggggatc gagcccgggg tgggcgaaga actccagcat 7201 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa 7261 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg 7321 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag 7381 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat 7441 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc 7501 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata 7561 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catccgcgcc 7621 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc 7681 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg 7741 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg 7801 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg 7861 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga 7921 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttggagttc attcagggca 7981 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg 8041 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc 8101 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat 8161 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc 8221 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc 8281 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa 8341 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca 8401 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag 8461 cagcccttgc gccctgagtg cttgcggcag cgtgagagct catgagcgga gaacgagatg 8521 acgttggagg ggcaaggtcg cgctgattgc tggggcaaca cgtggagcgg atcggggatt 8581 gtctttcttc agctcgctga tgatatgctg acgctcaatg ccgtttggcc tccgactaac 8641 gaaaatcccg catttggacg gctgatccga ttggcacggc ggacggcgaa tggcggagca 8701 gacgctcgtc cgggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtatcgggc 8761 cctggccagc tagctagagt cgacctgcag gtccccgggg atcggtcttg ccttgctcgt 8821 cggtgatgta cttcaccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt 8881 gcttggcaat cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc 8941 ctcgggcgga ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc 9001 gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc 9061 cagagtttca gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg 9121 acgtgctcat agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag 9181 gccgacaggc tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga 9241 aggcagtaca ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc 9301 cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg 9361 ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg 9421 cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga 9481 accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct 9541 ataatgaccc cgaagcaggg ttatgcagcg gaaaagatcc gtcgacctgc aggcatgcaa 9601 gctctagcga ttccagacgt cccgaaggcg tggcgcggct tccccgtgcc ggagcaatcg 9661 ccctgggtgg gttacacgac gcccctctat ggcccgtact gacggacaca ccgaagcccc 9721 ggcggcaacc ctcagcggat gccccggggc ttcacgtttt cccaggtcag aagcggtttt 9781 cgggagtagt gccccaactg gggtaacctt tgagttctct cagttggggg cgtagggtcg 9841 ccgacatgac acaaggggtt gtgaccgggg tggacacgta cgcgggtgct tacgaccgtc 9901 agtcgcgcga gcgcgaaaat tcgagcgcag caagcccagc gacacagcgt agcgccaacg 9961 aagacaaggc ggccgacctt cagcgcgaag tcgagcgcga cgggggccgg ttcaggttcg 10021 tcgggcattt cagcgaagcg ccgggcacgt cggcgttcgg gacggcggag cgcccggagt 10081 tcgaacgcat cctgaacgaa tgccgcgccg ggcggctcaa catgatcatt gtctatgacg 10141 tgtcgcgctt ctcgcgcctg aaggtcatgg acgcgattcc gattgtctcg gaattgctcg 10201 ccctgggcgt gacgattgtt tccactcagg aaggcgtctt ccggcaggga aacgtcatgg 10261 acctgattca cctgattatg cggctcgacg cgtcgcacaa agaatcttcg ctgaagtcgg 10321 cgaagattct cgacacgaag aaccttcagc gcgaattggg cgggtacgtc ggcgggaagg 10381 cgccttacgg cttcgagctt gtttcggaga cgaaggagat cacgcgcaac ggccgaatgg 10441 tcaatgtcgt catcaacaag cttgcgcact cgaccactcc ccttaccgga cccttcgagt 10501 tcgagcccga cgtaatccgg tggtggtggc gtgagatcaa gacgcacaaa caccttccct 10561 tcaagccggg cagtcaagcc gccattcacc cgggcagcat cacggggctt tgtaagcgca 10621 tggacgctga cgccgtgccg acccggggcg agacgattgg gaagaagacc gcttcaagcg 10681 cctgggaccc ggcaaccgtt atgcgaatcc ttcgggcccc gcgtattgcg ggcttcgccg 10741 ctgaggtgat ctacaagaag aagccggacg gcacgccgac cacgaagatt gagggttacc 10801 gcattcagcg cgacccgatc acgctccggc cggtcgagct tgattgcgga ccgatcatcg 10861 agcccgctga gtggtatgag cttcaggcgt ggttggacgg cagggggcgc ggcaaggggc 10921 tttcccgggg gcaagccatt ctgtccgcca tggacaagct gtactgcgag tgtggcgccg 10981 tcatgacttc gaagcgcggg gaagaatcga tcaaggactc ttaccgctgc cgtcgccgga 11041 aggtggtcga cccgtccgca cctgggcagc acgaaggcac gtgcaacgtc agcatggcgg 11101 cactcgacaa gttcgttgcg gaacgcatct tcaacaagat caggcacgcc gaaggcgacg 11161 aagagacgtt ggcgcttctg tgggaagccg cccgacgctt cggcaagctc actgaggcgc 11221 ctgagaagag cggcgaacgg gcgaaccttg ttgcggagcg cgccgacgcc ctgaacgccc 11281 ttgaagagct gtacgaagac cgcgcggcag gcgcgtacga cggacccgtt ggcaggaagc 11341 acttccggaa gcaacaggca gcgctgacgc tccggcagca aggggcggaa gagcggcttg 11401 ccgaacttga agccgccgaa gccccgaagc ttccccttga ccaatggttc cccgaagacg 11461 ccgacgctga cccgaccggc cctaagtcgt ggtgggggcg cgcgtcagta gacgacaagc 11521 gcgtgttcgt cgggctcttc gtagacaaga tcgttgtcac gaagtcgact acgggcaggg 11581 ggcagggaac gcccatcgag aagcgcgctt cgatcacgtg ggcgaagccg ccgaccgacg 11641 acgacgaaga cgacgcccag gacggcacgg aagacgtagc ggcgtagcga gacacccggg 11701 aagcctgatc tacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc 11761 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg 11821 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcga ctagt //