质粒名称: | p46Cpf1-OP2 |
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出品公司: | 爱迪基因 |
目录编号: | 98592 |
存储实验室: | Qiong Wu |
载体骨架基本信息 |
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载体名称: | pKD46 |
载体出品公司: | Datsenko KA & Wanner BL (PMID:10829079) |
载体抗性: | 氯霉素 |
载体类型: | 细菌表达,CRISPR |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 30 ℃ |
插入基因信息 |
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插入基因名称: | FnCpf1 |
物种来源: | Francisella |
基因突变: | Codon optimized for E. coli |
启动子 | araBAD |
克隆方法: | 限制性内切酶 |
5' 测序引物 | pBAD-F (ATGCCATAGCATTTTTATCC) |
3' 测序引物 | rrnB-T1-term-Rev (GAAAGGCCCAGTCTTTCGAC) |
文献引用方法 |
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材料和方法部分: | p46Cpf1-OP2 was a gift from Qiong Wu (Addgene plasmid # 98592 ; http://n2t.net/addgene:98592 ; RRID:Addgene_98592) |
买家导航
p46Cpf1OP2载体基本信息
p46Cpf1OP2载体图谱质粒图谱和多克隆位点信息
p46Cpf1OP2载体简介
p46Cpf1OP2载体序列
LOCUS p46Cpf1-OP2 10917 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Produces lambda Red and Cpf1 for recombination and selection. The plasmid is combined with a donor plasmid (with crRNA and template) for rapid genome editing in E.coli.. ACCESSION . VERSION . KEYWORDS p46Cpf1-OP2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10917) TITLE Qiong Wu CRISPR plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 10917) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10917 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(18..641) /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind 549..568 /label=Tet-R /note="Tetracycline resistance gene, reverse primer" promoter 660..715 /gene="tetR" /label=tetR/tetA promoters /note="overlapping promoters for bacterial tetR and tetA" protein_bind 696..714 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" RBS 733..744 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 758..1174 /codon_start=1 /gene="gam" /product="inhibitor of the host RecBCD nuclease in the lambda Red system" /label=Gam /note="protects the double-stranded DNA substrate for recombination" /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV" CDS 1180..1965 /codon_start=1 /gene="bet" /product="single-stranded DNA binding recombinase in the lambda Red system" /label=Beta /note="mediates pairing of a single-stranded DNA with a complementary target" /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA" CDS 1962..2642 /codon_start=1 /gene="exo" /product="5' to 3' double-stranded DNA exonuclease in the lambda Red system" /label=Exo /note="creates 3' single-stranded DNA tails" /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG EQWR" terminator 2643..2887 /label=-lambda tL3 terminator /note="transcription terminator tL3 from phage lambda" CDS complement(3026..3976) /codon_start=1 /gene="rep101 encoding an A56V mutation" /product="temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)" /label=Rep101(Ts) /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 4024..4246 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator 4767..4861 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(4872..5750) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 5777..6061 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 5984..6003 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 6083..6094 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" terminator 10015..10086 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 10102..10129 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(10155..10814) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 10729..10748 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(10815..10917) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 gtcgagcatg cgacgtctta agacccactt tcacatttaa gttgtttttc taatccgcat 61 atgatcaatt caaggccgaa taagaaggct ggctctgcac cttggtgatc aaataattcg 121 atagcttgtc gtaataatgg cggcatacta tcagtagtag gtgtttccct ttcttcttta 181 gcgacttgat gctcttgatc ttccaatacg caacctaaag taaaatgccc cacagcgctg 241 agtgcatata atgcattctc tagtgaaaaa ccttgttggc ataaaaaggc taattgattt 301 tcgagagttt catactgttt ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg 361 cgatgactta gtaaagcaca tctaaaactt ttagcgttat tacgtaaaaa atcttgccag 421 ctttcccctt ctaaagggca aaagtgagta tggtgcctat ctaacatctc aatggctaag 481 gcgtcgagca aagcccgctt attttttaca tgccaataca atgtaggctg ctctacacct 541 agcttctggg cgagtttacg ggttgttaaa ccttcgattc cgacctcatt aagcagctct 601 aatgcgctgt taatcacttt acttttatct aatctagaca tcattaattc ctaatttttg 661 ttgacactct atcgttgata gagttatttt accactccct atcagtgata gagaaaagaa 721 ttcaaaagat ctaaagagga gaaaggctcg agaaaaaatg gatattaata ctgaaactga 781 gatcaagcaa aagcattcac taaccccctt tcctgttttc ctaatcagcc cggcatttcg 841 cgggcgatat tttcacagct atttcaggag ttcagccatg aacgcttatt acattcagga 901 tcgtcttgag gctcagagct gggcgcgtca ctaccagcag ctcgcccgtg aagagaaaga 961 ggcagaactg gcagacgaca tggaaaaagg cctgccccag cacctgtttg aatcgctatg 1021 catcgatcat ttgcaacgcc acggggccag caaaaaatcc attacccgtg cgtttgatga 1081 cgatgttgag tttcaggagc gcatggcaga acacatccgg tacatggttg aaaccattgc 1141 tcaccaccag gttgatattg attcagaggt ataaaacgaa tgagtactgc actcgcaacg 1201 ctggctggga agctggctga acgtgtcggc atggattctg tcgacccaca ggaactgatc 1261 accactcttc gccagacggc atttaaaggt gatgccagcg atgcgcagtt catcgcatta 1321 ctgatcgttg ccaaccagta cggccttaat ccgtggacga aagaaattta cgcctttcct 1381 gataagcaga atggcatcgt tccggtggtg ggcgttgatg gctggtcccg catcatcaat 1441 gaaaaccagc agtttgatgg catggacttt gagcaggaca atgaatcctg tacatgccgg 1501 atttaccgca aggaccgtaa tcatccgatc tgcgttaccg aatggatgga tgaatgccgc 1561 cgcgaaccat tcaaaactcg cgaaggcaga gaaatcacgg ggccgtggca gtcgcatccc 1621 aaacggatgt tacgtcataa agccatgatt cagtgtgccc gtctggcctt cggatttgct 1681 ggtatctatg acaaggatga agccgagcgc attgtcgaaa atactgcata cactgcagaa 1741 cgtcagccgg aacgcgacat cactccggtt aacgatgaaa ccatgcagga gattaacact 1801 ctgctgatcg ccctggataa aacatgggat gacgacttat tgccgctctg ttcccagata 1861 tttcgccgcg acattcgtgc atcgtcagaa ctgacacagg ccgaagcagt aaaagctctt 1921 ggattcctga aacagaaagc cgcagagcag aaggtggcag catgacaccg gacattatcc 1981 tgcagcgtac cgggatcgat gtgagagctg tcgaacaggg ggatgatgcg tggcacaaat 2041 tacggctcgg cgtcatcacc gcttcagaag ttcacaacgt gatagcaaaa ccccgctccg 2101 gaaagaagtg gcctgacatg aaaatgtcct acttccacac cctgcttgct gaggtttgca 2161 ccggtgtggc tccggaagtt aacgctaaag cactggcctg gggaaaacag tacgagaacg 2221 acgccagaac cctgtttgaa ttcacttccg gcgtgaatgt tactgaatcc ccgatcatct 2281 atcgcgacga aagtatgcgt accgcctgct ctcccgatgg tttatgcagt gacggcaacg 2341 gccttgaact gaaatgcccg tttacctccc gggatttcat gaagttccgg ctcggtggtt 2401 tcgaggccat aaagtcagct tacatggccc aggtgcagta cagcatgtgg gtgacgcgaa 2461 aaaatgcctg gtactttgcc aactatgacc cgcgtatgaa gcgtgaaggc ctgcattatg 2521 tcgtgattga gcgggatgaa aagtacatgg cgagttttga cgagatcgtg ccggagttca 2581 tcgaaaaaat ggacgaggca ctggctgaaa ttggttttgt atttggggag caatggcgat 2641 gacgcatcct cacgataata tccgggtagg cgcaatcact ttcgtctact ccgttacaaa 2701 gcgaggctgg gtatttcccg gcctttctgt tatccgaaat ccactgaaag cacagcggct 2761 ggctgaggag ataaataata aacgaggggc tgtatgcaca aagcatcttc tgttgagtta 2821 agaacgagta tcgagatggc acatagcctt gctcaaattg gaatcaggtt tgtgccaata 2881 ccagtagaaa cagacgaaga atccatggag ctcatatggt accggtatgg acagttttcc 2941 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact 3001 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta 3061 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt 3121 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat 3181 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta 3241 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc 3301 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca 3361 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa 3421 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc 3481 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat 3541 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga 3601 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt 3661 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat 3721 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag 3781 cattttccct actgatgttc atcatctgaa cgtattggtt ataagtgaac gataccgtcc 3841 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta 3901 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa 3961 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga 4021 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat 4081 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta 4141 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag 4201 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag 4261 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt 4321 aaaaccctaa aggcttaagt agcaccctcg caagctcggt tgcggccgca atcgggcaaa 4381 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga 4441 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg 4501 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt 4561 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca 4621 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca 4681 gactggctaa tgcacccagt aaggcagcgg tatcatcaac ggggtctgac gctcagtgga 4741 acgaaaactc acgttaaggg cctagggact cctgttgata gatccagtaa tgacctcaga 4801 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa 4861 tcatcgattt attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca 4921 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa gtagagttga 4981 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc 5041 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc 5101 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg 5161 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga 5221 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc 5281 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg 5341 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc 5401 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag 5461 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct 5521 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc 5581 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg 5641 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg 5701 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata 5761 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc 5821 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg 5881 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc 5941 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta 6001 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca 6061 tgtgcacgaa ttcaaaagat ctaaagagga gaaaggatct atgtctatct accaggaatt 6121 cgttaacaaa tactctctgt ctaaaaccct gcgtttcgaa ctgatcccgc agggtaaaac 6181 cctggaaaac atcaaagctc gtggtctgat cctggacgac gaaaaacgtg ctaaagacta 6241 caaaaaagct aaacagatca tcgacaaata ccaccagttc ttcatcgaag aaatcctctc 6301 gtctgtctgc atcagcgaag acctgctgca gaactactcg gacgtgtact tcaaactaaa 6361 aaaatctgac gacgacaacc tgcagaaaga cttcaaatct gctaaagaca ccatcaaaaa 6421 acagatctct gaatacatca aagactctga aaaattcaaa aacctgttca accagaacct 6481 gatcgacgct aaaaaaggtc aggaatctga cctgatcctg tggctgaaac agtctaaaga 6541 caacggtatc gaactgttca aagctaactc tgacatcacc gacatcgacg aagctctgga 6601 aatcatcaaa tctttcaaag gttggaccac ctacttcaaa ggtttccacg aaaaccgtaa 6661 aaacgtttac tcttctaacg acatcccgac ctctatcatc taccgtatcg ttgacgacaa 6721 cctgccgaaa ttcctggaaa acaaagctaa atacgaatct ctgaaagaca aagctccgga 6781 agctatcaac tacgaacaga tcaaaaaaga cctggctgaa gaactgacct tcgacatcga 6841 ctacaaaacc tctgaagtta accagcgtgt tttctctctg gacgaagttt tcgaaatcgc 6901 taacttcaac aactacctga accagtctgg tatcaccaaa ttcaacacca tcatcggtgg 6961 taaattcgtt aacggtgaaa acaccaaacg taaaggtatc aacgaataca tcaacctgta 7021 ctctcagcag atcaacgaca aaaccctgaa aaaatacaaa atgtctgttc tgttcaaaca 7081 gatcctgtct gacaccgaat ctaaatcttt cgttatcgac aaactggaag acgactctga 7141 cgttgttacc accatgcagt ctttctacga acagatcgct gctttcaaaa ccgttgaaga 7201 aaaatctatc aaagaaaccc tgtctctgct gttcgacgac ctgaaagctc agaaactgga 7261 cctgtctaaa atctacttca aaaacgacaa atctctgacc gacctctctc agcaggtatt 7321 cgacgactac agcgttatcg gtaccgctgt tctggaatac atcacccagc agatcgctcc 7381 gaaaaacctg gacaacccgt ctaaaaaaga acaggaactg atcgctaaaa aaaccgaaaa 7441 agctaaatac ctgtctctgg aaaccatcaa actggctctg gaagaattca acaaacaccg 7501 tgacatcgac aaacagtgcc gtttcgaaga aatcctggct aacttcgctg ctatcccgat 7561 gatcttcgac gaaatcgctc agaacaaaga caacctggct cagatctcta tcaaatacca 7621 gaaccagggt aaaaaagacc tgctgcaggc ttctgctgaa gacgacgtta aagctatcaa 7681 agacctgctg gaccagacca acaacctgct gcacaaactg aaaatcttcc acatctctca 7741 gtctgaagac aaagctaaca tcctggacaa agacgaacac ttctacctgg ttttcgaaga 7801 atgctacttc gaactggcta acatcgttcc gctgtacaac aaaatccgta actacatcac 7861 ccagaaaccg tactctgacg aaaaattcaa actgaacttc gaaaactcta ccctggctaa 7921 cggttgggac aaaaacaaag aaccggacaa caccgctatc ctgttcatca aagacgacaa 7981 atactacctg ggtgttatga acaaaaaaaa caacaaaatc ttcgacgaca aagctatcaa 8041 agaaaacaaa ggtgaaggtt acaaaaaaat cgtttacaaa ctgctgccgg gtgctaacaa 8101 aatgctgccg aaagttttct tctctgctaa atctatcaaa ttctacaacc cgtctgaaga 8161 catcctgcgt atccgtaacc actctaccca caccaaaaac ggttctccgc agaaaggtta 8221 cgaaaaattc gaattcaaca tcgaagactg ccgtaaattc atcgacttct acaaacagtc 8281 tatctctaaa cacccggaat ggaaagactt cggtttccgt ttctctgaca cccagcgtta 8341 caactctatc gacgaattct accgtgaagt tgaaaaccag ggttacaaac tgaccttcga 8401 aaacatctct gaatcttaca tcgactctgt tgttaaccag ggtaaactgt acctgttcca 8461 gatctacaac aaagacttct ctgcttactc taaaggtcgt ccgaacctgc acaccctgta 8521 ctggaaagct ctgttcgacg aacgtaacct gcaggacgtt gtttacaaac tgaacggtga 8581 agctgaactg ttctaccgta aacagtctat cccgaaaaaa atcacccacc cggctaaaga 8641 agctatcgct aacaaaaaca aagacaaccc gaaaaaagaa tctgttttcg aatacgacct 8701 gatcaaagac aaacgtttca ccgaagacaa attcttcttc cactgcccga tcaccatcaa 8761 cttcaaatct tctggtgcta acaaattcaa cgacgaaatc aacctgctgc tgaaagaaaa 8821 agctaacgac gttcacatcc tgtctatcga ccgtggtgaa cgtcacctgg cttactacac 8881 cctggttgac ggtaaaggta acatcatcaa acaggacacc ttcaacatca tcggtaacga 8941 ccgtatgaaa accaactacc acgacaaact ggctgctatc gaaaaagacc gtgactctgc 9001 tcgtaaagac tggaaaaaaa tcaacaacat caaagaaatg aaagaaggtt acctgtctca 9061 ggttgttcac gaaatcgcta aactggttat agaatacaat gcgatagttg tatttgaaga 9121 cctgaacttc ggcttcaaac gtggtcgttt caaagttgaa aaacaggttt accagaaact 9181 ggaaaaaatg ctgatcgaaa aactgaacta cctggttttc aaagacaacg aattcgacaa 9241 aaccggtggt gttctgcgtg cttaccagct gaccgctccg ttcgaaacct tcaaaaaaat 9301 gggtaaacag accggtatca tctactacgt tccggctggt ttcacctcta aaatctgccc 9361 tgttactggc ttcgttaacc agctctaccc gaaatacgaa tctgtttcta aatctcagga 9421 attcttctct aaattcgaca aaatctgcta caacctggac aaaggttact tcgaattctc 9481 tttcgactac aaaaacttcg gtgacaaagc tgctaaaggt aaatggacca tcgcttcttt 9541 cggttctcgt ctgatcaact tccgtaactc tgacaaaaac cacaactggg acacccgtga 9601 agtttacccg accaaagaac tggaaaaact gctgaaagac tactctatcg aatacggtca 9661 cggtgaatgc atcaaagctg ctatctgcgg tgaatctgac aaaaaattct tcgctaaact 9721 gacctctgtt ctgaacacca tcctgcagat gcgtaactct aaaaccggta ccgaactgga 9781 ctacctgatc tctccggttg ctgacgttaa cggtaacttc ttcgactctc gtcaggctcc 9841 gaaaaacatg ccgcaggacg ctgacgctaa cggtgcttac cacatcggtc tgaaaggtct 9901 gatgctgctg ggtcgtatca aaaacaacca ggaaggtaaa aaactgaacc tggttatcaa 9961 aaacgaagaa tacttcgaat tcgttcagaa ccgtaacaac taataaccag gcatcaaata 10021 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac 10081 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgg tgcacaagcg 10141 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta 10201 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc 10261 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag 10321 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag 10381 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac 10441 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag 10501 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc 10561 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg 10621 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt 10681 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga 10741 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg 10801 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg 10861 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatca //