质粒名称: | pBFC0625 |
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出品公司: | Addgene |
目录编号: | 186691 |
存储实验室: | Jennifer Doudna |
载体骨架基本信息 |
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载体名称: | R6K conditional origin, bla, RP4 oriT |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | 12613 bp |
载体修饰: | -- |
载体类型: | 细菌表达 |
筛选标记: | - |
载体抗性: | 氨苄青霉素和庆大霉素,100和10μg/mL |
生长温度: | 37 ℃ |
宿主菌: | EC100D pir+ |
拷贝数: | 低拷贝 |
插入基因信息 |
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插入基因名称: | tnsA tnsB tnsC tnsD tniQ cas7 cas6 Vc_lacZ_α_1 Spacer crRNA Tn7 transposon GmR+sfGFP VcDART Cargo |
物种来源: | Vibrio cholerae |
文献引用方法 |
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材料和方法部分: | pBFC0625 was a gift from Jennifer Doudna (Addgene plasmid # 186691 ; http://n2t.net/addgene:186691 ; RRID:Addgene_186691) |
参考文献部分: | Species- and site-specific genome editing in complex bacterial communities. Rubin BE, Diamond S, Cress BF, Crits-Christoph A, Lou YC, Borges AL, Shivram H, He C, Xu M, Zhou Z, Smith SJ, Rovinsky R, Smock DCJ, Tang K, Owens TK, Krishnappa N, Sachdeva R, Barrangou R, Deutschbauer AM, Banfield JF, Doudna JA. Nat Microbiol. 2022 Jan;7(1):34-47. doi: 10.1038/s41564-021-01014-7. Epub 2021 Dec 6. 10.1038/s41564-021-01014-7 PubMed 34873292 |
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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 pBFC0625 12613 bp ds-DNA circular SYN 09-SEP-2022 DEFINITION VcDART under control of Lac promoter, Vc_lacZ_alpha_1 gRNA, GmR+sfGFP cargo. ACCESSION . VERSION . KEYWORDS pBFC0625 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12613) AUTHORS Rubin BE, Diamond S, Cress BF, Crits-Christoph A, Lou YC, Borges AL, Shivram H, He C, Xu M, Zhou Z, Smith SJ, Rovinsky R, Smock DCJ, Tang K, Owens TK, Krishnappa N, Sachdeva R, Barrangou R, Deutschbauer AM, Banfield JF, Doudna JA TITLE Species- and site-specific genome editing in complex bacterial communities. JOURNAL Nat Microbiol. 2022 Jan;7(1):34-47. doi: 10.1038/s41564-021-01014-7. Epub 2021 Dec 6. PUBMED 34873292 REFERENCE 2 (bases 1 to 12613) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 9, 2022 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..12613 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 270..341 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 357..384 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind 395..414 /label=T7 /note="T7 promoter, forward primer" promoter 395..413 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" oriT 557..665 /note="incP origin of transfer" promoter 858..962 /gene="bla" /label=AmpR promoter CDS 963..1823 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1181..1200) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1859..2247 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" promoter 2397..2427 /label=lac promoter /note="promoter for the E. coli lac operon" RBS 2428..2439 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 4524..4535 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 11031..11059 /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" CDS 11248..11781 /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" RBS 12140..12151 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 12158..258 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" ORIGIN 1 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag 61 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct 121 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac 181 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt 241 atggatgaac tgtacaaatg accaggcatc aaataaaacg aaaggctcag tcgaaagact 301 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct 361 caccttcggg tgggcctttc tgcgtttata gctgtaatac gactcactat aggggataag 421 agacggctca acgcccgtct cacagcgcaa tatcaattta tgggtgtgat aattatcaat 481 ttatgggtgt aattatcatt ttatggttgt atcaacaagg aggactttgg caggtgcctg 541 caggattctt gcttatcggc cagcctcgca gagcaggatt cccgttgagc accgccaggt 601 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc 661 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc 721 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc 781 agggttatgc agcggaaaag aatgtcatga taataatggt ttcttagacg tcaggtggca 841 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 901 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 961 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 1021 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 1081 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 1141 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 1201 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 1261 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 1321 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 1381 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 1441 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 1501 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 1561 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 1621 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggct 1681 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 1741 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 1801 cctcactgat taagcattgg taactgtcag acccggccac gatgcgtccg gcgtagagga 1861 tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac 1921 taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta 1981 acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca 2041 tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt 2101 ttataagaaa aaaaagaata tataaggctt ttaaagcttt taaggtttaa cggttgtgga 2161 caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag 2221 tgacacagga acacttaacg gctgacatgg gcaagtttac tcatatatac tttagattga 2281 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 2341 gaccaaaatc ccttaacgtg agttttcgtt ccactgacac ctgcccgaac catatcttta 2401 cactttatgc ttccggctcg tatgttgaaa gaggagaaag gatcaatggc gacaagttta 2461 cctacgccct cagcaattac gacttcggcg ttagagtatg cattccatac tcccgctcgc 2521 aatctaacga aatctcgcgg aaaaaacatt catcgttatg tcagtgtaaa gatgagtaag 2581 aggattacgg tagaatctac tctagagtgt gatgcctgct atcactttga ttttgagcca 2641 agtattgttc gcttttgcgc tcaaccgatt cgatttttat attatctcaa tggtcagtct 2701 cactcctatg ttcctgactt tctagttcaa tttgatacca acgagtttgt tctatatgaa 2761 gtaaagtcag cttatgctaa gaacaaacct gattttgatg ttgaatggga ggcgaaagta 2821 aaagcagcaa ctgaactagg gctagaattg gagcttgttg aagagagtga tattagggat 2881 acggttgtat taaataatct taagcgcatg catcgttatg cttcgaaaga tgagctgaat 2941 aacgtacata actctctctt aaaaataata aagtacaatg gcgcccaatc tgcaagatgc 3001 ttgggagaac agttgggttt aaaaggccga actgttttac caattttgtg cgatttgctg 3061 tcaaggtgtt tactcgatac acgtttggat aagcctctat ctcttgaatc tcgatttgag 3121 ttggccagtt atggctaaga aagggttctc aagtttccat agaaaagcag tctcttccca 3181 agatacgctt gaatcgatag agcttgtctc tagcgctaat tgtctagaaa gtgttacgta 3241 tcaagatata tcagcatttc ccgaaacaat tgcggtagag attaatttcc gattaagcat 3301 tcttcgcttt ttagcgcgaa agtgcgaaac cattgtggcc aaatcaattg aaccacatcg 3361 tgtagagcta cagcaaaact atagtagaaa aatacccagt gcaataacga tatatcgatg 3421 gtggcttgct tttcgaaaat cagactacaa ccccattagc ttagcaccta atatcaagga 3481 tagaggtaat agagagacaa aagtgtcaac agttgttgat tctattatgg aacaggcagt 3541 tgaaagagtt atatctggac gaaaagtcaa tgttagctct gcatataaac gtgttcgacg 3601 aaaagttcgt caatacaatc taactcatgg aacgaaatac acgtatccta agtacgaatc 3661 tgtaagaaag cgagtaaaaa agaaaacccc atttgagtta ttagccgcag ggaaagggga 3721 gagagtagct aagagagagt ttcgccgaat gggaaaaaag atcctcacgt ctagcgtgct 3781 agagagggtt gaaatagatc acactgtcgt tgaccttttt gcagtacacg aagagtatcg 3841 aatcccattg ggccgacctt ggcttactca attggttgat tgttacagta aagctgtaat 3901 cggtttttat ttaggtttcg agcctcctag ctatgtgtcg gtttcccttg cacttaagaa 3961 tgcaatacaa cgcaaagatg acttaatctc ctcgtatgaa tcgatcgaga atgaatggct 4021 atgttatggc atcccagacc tactcgtaac tgataatggt aaagagtttt tgtcgaaagc 4081 atttgatcaa gcatgtgaat cactattgat caatgtgcat caaaataaag ttgaaacgcc 4141 cgacaacaaa cctcatgttg aacgtaacta cgggactatt aatacttctc tgttagacga 4201 tttacctggg aaatccttca gccagtacct tcaaagagaa gggtacgact ctgtgggaga 4261 agctaccctt acactcaatg agattagaga aatttactta atttggttgg tggatattta 4321 tcataaaaaa cccaatcaga gaggcactaa ttgtcctaat gttgcgtgga aaaagggttg 4381 tcaagaatgg gaaccagagg agttctctgg ttctaaagac gaattagact ttaaatttgc 4441 tattgttgat tacaaacaac ttactaaagt agggataact gtctacaaag aactgagtta 4501 tagcaatgac cgtttagctg aatatagagg gaagaaagga aaccataaag ttcagttcaa 4561 gtataaccct gagtgtatgg cagttatttg ggtgttggat gaggatatga atgagtactt 4621 tacagttaat gcgattgact acgaatatgc aagtagagta tcactttggc aacataaata 4681 taacatgaaa tatcaagcag aactaaattc agcagaatat gatgaggaca aggaaattga 4741 tgcagaaata aaaattgaag aaatcgcaga tcgttcaatt gttaagacta acaaaatcag 4801 agctcggagg cgtggcgcta ggcatcaaga gaatagcgca agggctaagt caatcagtaa 4861 tgcgaacccg gcctcgatac aaaaacatga agatgaaatc gttagtgcag ataatgacga 4921 ttgggatatt gattatgtct gagaaatcgt caaatgagtg aaacgcgtga ggctcgtata 4981 tcaagagcta aaagggcatt tgtatccaca ccgagcgtta ggaaaatctt gagttacatg 5041 gatagatgta gagatctatc agacctagag tctgagccta catgcatgat ggtctatggt 5101 gcttcgggtg taggtaaaac gaccgtcatc aagaaatact taaatcaaaa cagaagagag 5161 tccgaagccg ggggcgatat aataccggtt ttgcatattg agttgccaga caatgcgaag 5221 ccagtagatg cagcaaggga attgctggtt gaaatgggtg acccgctagc actttatgaa 5281 actgacttag ctagattgac gaaaagactg actgaattaa tccctgcggt cggcgtgaag 5341 ctgattatta tcgatgagtt ccaacatttg gtggaagaaa ggtcaaatcg ggttcttacc 5401 caagtaggta attggctaaa aatgatactt aacaaaacga aatgtccaat tgttatattt 5461 ggtatgccat actcaaaagt tgtactgcaa gcaaactcgc aacttcacgg gcgattttcc 5521 attcaggttg aactgcgccc ctttagctac cagggaggta gaggtgtatt taaaactttt 5581 ttggaatacc ttgataaagc cctacctttt gaaaaacagg ctggcttagc caacgaaagt 5641 ttgcagaaaa aattgtatgc attctctcag ggaaacatgc gttcgttgag aaaccttatt 5701 tatcaagcat ctatcgaagc aattgataat cagcatgaaa cgataaccga agaagatttc 5761 gtttttgcat cgaagttgac atcgggcgat aaacccaact catggaaaaa tccttttgag 5821 gagggtgttg aggtaacaga agatatgtta cgaccgccac caaaagatat tggttgggag 5881 gactatttga gacattcaac cccgagagtg agtaaaccag gtagaaataa aaactttttc 5941 gaataaccta ggctgctgcc accgctgagc aataaccatc ttagtatatt agttaagtat 6001 aagaaggaga tatacatatg tttttgcaaa gacctaaacc ttacagcgat gaaagtttag 6061 aaagtttctt tatccgagtg gctaacaaaa atggctacgg tgatgtccat cgcttcctag 6121 aagccactaa acgattcctt caagacattg accataatgg ctatcaaacc tttccgactg 6181 atataactcg gataaaccca tactcagcta aaaacagttc cagcgcacga actgcgtcat 6241 tcctgaaact tgcacaattg acatttaatg aaccgccaga gctacttggg ttggcaatta 6301 acagaacaaa catgaaatac tcgccgtcaa ctagcgcggt tgttagaggt gcagaagtct 6361 ttcctcgcag tttactacgg acgcactcca tcccctgctg tcctttgtgt ctgcgagaaa 6421 atggctacgc ctcctacctt tggcactttc aggggtacga atactgccac agccataacg 6481 tacctttaat taccacttgt agctgtggta aggagtttga ctaccgagta tctgggttaa 6541 agggcatttg ctgcaaatgc aaggagccta tcaccttaac cagcagggag aacggtcatg 6601 aggcagcgtg tactgtttca aactggcttg ctggccatga atctaaacct ctgccaaatc 6661 ttcctaaaag ctaccgatgg ggtttagttc attggtggat gggtattaaa gatagcgagt 6721 tcgatcactt ttcgttcgtt caatttttct caaactggcc aaggtcattc cactcgataa 6781 tcgaagatga agtagagttc aaccttgagc atgctgttgt cagcacgtct gaattacgac 6841 taaaagatct tcttggtcga ttgtttttcg gttcaattcg gttacctgag cggaatcttc 6901 aacacaatat catccttggt gagcttctct gctatttaga aaatcgttta tggcaagaca 6961 agggattaat cgccaacctc aaaatgaacg cgttagaggc gactgtaatg ttaaattgta 7021 gcctcgatca gattgcatca atggttgaac aacgcatctt gaagccaaat cgaaaaagca 7081 agcccaacag ccctcttgat gttaccgatt atctatttca tttcggcgat attttctgtc 7141 tttggttagc tgagttccaa agcgatgagt ttaaccgttc gttttatgtg tcgaggtggt 7201 aaatgcaaac tctgaaagaa ctaatcgcat ccaatcctga cgacttaaca actgaactta 7261 agagagcatt tcgtccactc acaccgcata ttgcaattga tggtaatgaa cttgacgcac 7321 tgacgatatt agtcaattta accgataaga ctgatgatca gaaagacctg ctcgatcgag 7381 ccaaatgcaa gcaaaaactt cgagatgaaa aatggtgggc tagctgcata aattgcgtta 7441 attacagaca aagccataac ccaaaattcc cggatatacg ttctgaaggc gtgattcgaa 7501 cccaagccct gggtgaatta ccgagtttcc tactctcgtc ctcaaaaatc ccaccatacc 7561 attggtcata tagccatgat tcaaaatacg tcaacaaaag cgcattcctc accaatgagt 7621 tttgttggga tggtgagatc tcatgtttgg gtgagcttct taaagatgca gatcacccac 7681 tttggaatac tttgaaaaag ttaggttgtt ctcaaaaaac ttgcaaagca atggcaaaac 7741 aactagctga tattactctc acgactatca atgtcacact cgcaccaaat tacctgactc 7801 aaatatctct ccccgatagt gatacatcct acatttcact ctctcctgtt gcatcgctat 7861 cgatgcaaag ccactttcat cagaggctcc aagatgaaaa tcggcatagc gcgataacgc 7921 ggtttagccg aactaccaac atgggggtca ctgcgatgac atgtggcggc gcatttagaa 7981 tgttaaagtc tggcgctaag ttttccagcc cccctcatca ccgattaaac agtaaacgaa 8041 gttggttgac gtcagagcat gttcagtcat taaaacagta ccagcgcctc aacaaaagcc 8101 tcatacctga aaactctcgg attgcactcc gtagaaaata caaaatcgag cttcaaaata 8161 tggtcagatc ttggtttgca atgcaagacc atacccttga ttcgaacata cttatccaac 8221 atttgaatca tgacctatct tacttaggag ccacaaaacg ttttgcatac gatccagcga 8281 tgaccaagct ctttactgag cttttgaaac gagagttatc aaattcaatc aataatggtg 8341 agcaacacac taatggatcg tttttagtcc taccgaatat cagggtttgt ggcgcaacag 8401 ctttaagctc cccggtaacg gtggggattc catcacttac agctttcttt ggcttcgttc 8461 acgcatttga acggaatata aatcgcacca cctcatcgtt tcgtgttgaa tcctttgcga 8521 tatgcgtcca tcaactacat gtcgagaagc gaggtttgac agcagagttt gtggaaaaag 8581 gcgacgggac tatatccgct cccgcgaccc gggatgactg gcagtgtgat gtcgtattta 8641 gccttatttt gaacaccaac tttgctcaac atattgacca agatacgtta gttacatcac 8701 taccaaagcg attggctcgg ggttcagcaa aaattgcgat tgatgacttt aaacatatca 8761 actcattctc gacattagaa acagcgatcg aatctctgcc aatagaagct ggtaggtggt 8821 tatcacttta cgcacagtca aacaataatc taagtgatct attagcagcc atgacagagg 8881 accatcagct catggcaagc tgcgtcggtt accacttgtt agaagaaccc aaagataaac 8941 caaactccct cagaggttac aaacacgcta tcgccgagtg catcattgga ctcattaact 9001 caatcacctt tagctcagag actgatccca acacaatctt ttggtcgcta aagaactatc 9061 aaaactacct agtggtacag ccaaggagta tcaacgatga aactaccgac aaatctagcc 9121 tatgagcgct ctatcgaccc atcagatgtc tgtttttttg tcgtctggcc cgatgataga 9181 aaaacacctt taacctacaa ttctcgtact ctgctcgggc aaatggaagc ggcatcatta 9241 gcctatgatg tctcaggtca accaataaaa agtgccaccg ctgaggcgtt agctcaaggg 9301 aaccctcatc aagttgattt ctgccacgtt ccatacggtg cgagtcatat tgaatgcagt 9361 ttctccgttt cgttttcttc tgaactacgt caaccatata agtgtaactc aagcaaagtt 9421 aaacaaacgc tagtgcaatt agtcgagctc tacgaaacga aaatcggctg gactgagcta 9481 gcaacccgat atttgatgaa catttgcaac ggtaaatggc tgtggaaaaa tacccgtaaa 9541 gcctattgct ggaacattgt acttacacct tggccttgga acggggaaaa ggttggattt 9601 gaagatattc gtactaacta cacctcacgg caagacttta aaaataataa aaattggtct 9661 gctatagttg aaatgatcaa aaccgcattt tctagtactg atgggctggc gatatttgaa 9721 gtcagggcca ccttgcactt gccaacgaat gctatggtgc ggccaagcca agttttcaca 9781 gaaaaagaaa gtggcagtaa aagtaaatct aaaactcaaa acagtcgagt ttttcagagt 9841 acaactattg atggtgaacg atcgccaata ctaggggcct ttaaaacggg agcagctatt 9901 gcaaccattg acgactggta tcctgaagcc actgagccac taagggtcgg acggtttggg 9961 gttcatcgcg aagatgtcac ttgctaccgt catccgtcta ccggaaaaga ttttttctcg 10021 atattacaac aagcagagca ctatattgaa gtgttgagcg ccaacaaaac tcccgctcaa 10081 gaaactatca acgacatgca ctttttaatg gctaacctga ttaagggtgg gatgttccag 10141 cataaaggag actgactgtg aaatggtatt ataagacaat cacctttctg ccagagttgt 10201 gcaacaacga gtcactggct gcaaagtgtc tccgcgttct gcatggattt aactatcagt 10261 atgagacacg aaatatcggc gtttcatttc cgctttggtg tgatgcaacg gttggaaaaa 10321 agatttcatt tgtcagcaag aacaagatag aactcgactt actacttaaa caacactatt 10381 tcgtccaaat ggaacaactt caatactttc atatatccaa cactgttctc gtcccagaag 10441 attgtacata cgtttccttt agacgctgtc aatctataga taagctcaca gcagcagggc 10501 tggcaaggaa aatcagacgc ctggagaaac gtgctctatc tagaggcgag caatttgacc 10561 catcatcttt tgctcaaaaa gagcatactg caatagcgca ctaccactca cttggggagt 10621 ccagcaaaca gacgaaccgc aactttcgac tcaatatcag gatgctctcg gagcaacccc 10681 gtgaggggaa ctcgattttt agtagctatg gcttatcaaa ttcagaaaac tcgtttcagc 10741 ctgtaccctt aatctgacct aggctgctgc caccggtgaa ctgccgagta ggtagctgat 10801 aacccctttc gccagctggc gtaatagcga agagggtgaa ctgccgagta ggtagctgat 10861 aacccaaacc agatgtcaac acagctactg ttgatgcaac cataaagtga tatttaataa 10921 ttatttataa tcagcaactt aaccacaaaa caaccatata ttgatatctc acaaaacaac 10981 cataagttga tatttttcag ccaggggtcc ccaataatta cgatttaaat ttgacataag 11041 cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa 11101 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact 11161 gtttttttgt acagcctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc 11221 gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag 11281 ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacgtgtagg 11341 ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc 11401 ggtgacgtag ccacctactc ccaacatcag ccggattccg attacctcgg gaacttgctc 11461 cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc 11521 gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg aaatctatat ctatgatctc 11581 gcagtctccg gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag 11641 catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat 11701 cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc 11761 gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag 11821 ttgttcggta aattggacaa cggtccgcgc gttgtccttt tccgctgcat aaccctgctt 11881 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt 11941 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgaaagtgaa 12001 acgtgatttc atgcgtcatt ttgaacattt tgtaaatctt atttaataat gtgtgcggca 12061 attcacattt aatttatgaa tgttttctta acatcgcggc aactcaagaa acggcaggtt 12121 cggatcttag ctactagaga aagaggagaa atactagatg cgtaaaggcg aggagctgtt 12181 cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc 12241 cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg 12301 tactactggt aaactgccgg ttccttggcc gactctggta acgacgctga cttatggtgt 12361 tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat 12421 gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac 12481 gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat 12541 tgactttaaa gaggacggca atatcctggg ccataagctg gaatacaatt ttaacagcca 12601 caatgtttac atc //