LOCUS Exported 10232 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses IgnaviCas9 in E. coli. ACCESSION . VERSION . KEYWORDS pIgnaviCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10232) AUTHORS Schmidt ST, Yu FB, Blainey PC, May AP, Quake SR TITLE Nucleic acid cleavage with a hyperthermophilic Cas9 from an uncultured Ignavibacterium. JOURNAL Proc Natl Acad Sci U S A. 2019 Oct 28. pii: 1904273116. doi: 10.1073/pnas.1904273116. PUBMED 31659048 REFERENCE 2 (bases 1 to 10232) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10232 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 61..80 /label=T7 /note="T7 promoter, forward primer" promoter 61..79 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 80..104 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 119..141 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 152..169 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 173..1270 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW YAVRTAVINAASGRQTVDEALKDAQT" primer_bind 1244..1267 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 1301..1321 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 5060..5080 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5157..5174 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(5227..5245) /label=T7 Term /note="T7 terminator, reverse primer" terminator 5241..5288 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 5325..5780 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5412..5431) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5622..5643 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(5872..6687) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 6595..6614 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 6679..6698 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 6809..7397 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7298..7317 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7551..7568 /label=L4440 /note="L4440 vector, forward primer" misc_feature 7583..7722 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(7648..7667) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7808..7830 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(7824..8015) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(8824..9906) /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 9868..9887 /label=LacI-R /note="LacI, reverse primer" promoter complement(9907..9984) /gene="lacI" /label=lacI promoter primer_bind 10190..10209 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat 61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 121 tgtttaactt taagaaggag atatacatat gcaccatcac catcaccatg gaaaaatcga 181 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 241 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 301 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 361 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 421 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 481 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 541 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 601 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 661 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 721 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 781 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 841 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 901 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 961 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 1021 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 1081 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ctatggaaaa 1141 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 1201 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 1261 cgcgcagact aattcgagct cgaacaacaa caacactagt gaaaacctgt atttccaggg 1321 agcagcctcg atgaaaaaag tattaggatt agatcttgga gtatcttcaa taggctgggc 1381 tttaattgac gaagatgata gaaaaataat gggcatgggt agtagaataa taccattaac 1441 aactgatgat aaagacgagt ttacaaaagg caatacgatt tctaagaatc agcaacgaac 1501 aattaaaaga actcaaagaa aaggatacga tcgttatcaa ttaagaaggc agaatttagt 1561 tttcgtgttg aaacaaaata atatgatgcc tgatattgaa ttagtaaatc ttccaaaact 1621 tgaattatgg aaactaagaa gtgatgcggt taataaaaaa atatctttga aagaattagg 1681 cagaatccta cttcacttaa atcaaaaaag aggttataaa agtagcagaa gtgaatcaaa 1741 tttggataag aaagataccg aatatgtagc aacagtaaaa aacagatatg aaagcctaaa 1801 agaaattggt ttaacaatag gacagaaatt ttttgaggaa ttatccaaaa acaattttta 1861 cagaataaaa gaacaggttt acccaagaga agcatatgtt gaagagtata ataaaataat 1921 gaagcatcaa caaaaacatt atccagaaaa tatttcggaa gaattaatta ataaaataag 1981 agacgaaata atttactatc aacgaaaact aaaatcgcaa aagggattgg tgtctgtttg 2041 cgagtttgaa ggattttgga taaagctaaa ttcaaatgga aaagaaaaag atttatttgt 2101 tggtccaaaa gtaactccta aaagttcacc attattccag gtaagtagaa tttgggaaac 2161 tatcaataac atatcaatta aaagaaagac tggtgaatcc attgaaatta cactggataa 2221 aaagaaagaa atttttgctt atatggataa aaatgaaaaa ttaagctatc cagaattatt 2281 aaaaatttta gggcttaaaa aagatgacgt atatggaaac aagaatttaa caaatgggtt 2341 gctgggcaac aaaataaaaa cagaaatgat gaagtgtatt tcagatattg ataagtattc 2401 tgatttattc cgattagaac ttgaaataaa agaattcgat gaagaggttt atttatatga 2461 tagaacaacc ggagaaataa taaattcaaa gaaaaaaaag aatataatag cagcaataga 2521 agaccaacca ttttacaagc tttggcatgt tgtttattca atacccgata aagaaacttg 2581 tcaaaaaata cttatgtcaa aatttggcat acaggaagaa gacgctgcta aattagcaac 2641 acttgatttt actaaacttg gtttttcgaa caaatcccac cgtgcaatta ggaaaatgct 2701 tccttatcta atggaagggg ataacgatta tatggcccgt tgttatgcgg gttatcatca 2761 cacaacaaca attacaaaac aagaaaactt ccaaagaaaa ctgttagata aattaaaaaa 2821 cttagaaaaa aatagcctgc gccagccgat agttgaaaaa attctaaatc agatgataaa 2881 tgttgtaaat gcaattatag acaaatatgg gaaaccggat gaaattagaa ttgaactagc 2941 cagagaatta aaacagagta gagaagaaag aaatgaagca tatagaaaca tgaatgaacg 3001 agaacgtgaa aataaaataa ttgaaaaaga gctttctgaa tttggacttc gtgcaacacg 3061 aaacaatatt atcaaatgga gattatatca cgaaattagc aacgaagaaa agaaacaaaa 3121 tgcaatttgc atttattgtg gcaaaccaat ttcctttact gctgcaatat taggtgaaga 3181 agttgaagtt gaacacataa taccaaggtc aaggttattt gacgattctc aaagcaataa 3241 aacactggca catagaaaat gcaatgcaga taagaaagac caaacagctt atgactttat 3301 gcgttcaaaa tctgatactg aatttaatga ttacgttgag cgaattaata ccctttataa 3361 aaatcatgta attggaaaaa cgaaaagaga taaactttta atgtctgaag aaaaaattcc 3421 tatggatttt attgacagac aattaagaca aacacaatac atctctaaaa aagcattaga 3481 gcttcttcag aatatctgtt ataatgtgtg ggcaacaagc ggaaatgtga ccgccgagtt 3541 gcgccatata tggggatggg atgaagtgct tgaaaatctt caattaccta agtatagaga 3601 aagtggatta atagaaatta ttgaagttgg agataaagat aataaacaaa aaaaggaaaa 3661 gataattgga tggaccaaaa gagacgatca tagacatcat gcaattgatg ctcttaccat 3721 cgcatgtacc aaacaaggat ttatccaacg ctttaataga ttaaatagtg ggaaagtacg 3781 aaacgatatg cttcaggaaa ttgaaaacgc caaacagaat tacgataaaa gaaaaaatct 3841 tttggagaac tatattcttt cttacagacc atttacaaca aaggaagttg aaagagaggc 3901 tgagaaaata cttgtatcat tcaaagccgg caaaaaggtt gcatctacag gcaaaagaaa 3961 aattaaaaaa gatggcaaaa aaataatcgc tcaaactggt attattattc caagaggacc 4021 attaagtgaa gaaagtgtct atggaaaaat aaaagtaatt gagaaggaaa aaccgttaaa 4081 atatttattt gaaaatccac acctcatatt taaaccaaat ataaaagcac ttgtagaaga 4141 aagactttac aaaaacaata acgaccctaa aagtgctata gcttcattaa aaaaagaacc 4201 tatttatctt gacaaagaga aaacaataaa attggaatac ggaacatgtt ataaagaaga 4261 agttgttata aaaaaaccac tacaagcttt gaacgagaag caagtagagg atattgttga 4321 ccctataata aaacaaaaga ttaaggatcg actggttaaa tttggtggca aagccaaaga 4381 agcatttaag gatttagaaa acgaacctat ttggtatgat gaggaaaaaa gaattccaat 4441 aaagaatgtt cgatggttta caggactttc agcaattgaa cctataagca aggatgagac 4501 cggaaaagaa attggatttg tcaaacctgg caataatcat catcttgcaa tatacattga 4561 tgaagaaggg aaaaaacaac ttagtatatg ttcattttgg catgctgtag aaagaaagaa 4621 atatgggttg cctgttataa taaaaaatcc gtcagaggtt gttgatttta tacttgcgga 4681 ggaaaatgaa gataaatatc cagaaagttt tctagaaaaa ttacccgctg ggaaatggac 4741 atttaaagaa agctttcaac aaaacgagat gtttgtactt ggaataagca aagaagcatt 4801 tgaagaagcc atttcgagaa atgattatag cttcttaagt aattacttat atcgtgttca 4861 aaagattgca atgataggca aacaaccaaa tattgttttt agacatcatc tcgaaactca 4921 gcttaaggat gacgcatacg ctaaaaaaag taatcgcttt tatttaatac aaagtatcgg 4981 ggcattagaa tcattatatc caataaaaat tttaattaat tgtttgggag aaattattac 5041 taataataaa taagggtcac ctaagaaaaa acgaaaagtt gagaccggtt aacgaggccg 5101 gtgctttgca ggatccgaat tcgagctccg tcgacaagct tgcggccgca ctcgagcacc 5161 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg 5221 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg 5281 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg 5341 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 5401 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 5461 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 5521 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 5581 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 5641 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 5701 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 5761 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt 5821 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa 5881 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt 5941 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc 6001 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt 6061 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg 6121 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg 6181 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc 6241 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg 6301 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa 6361 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt 6421 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac 6481 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg 6541 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg 6601 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca 6661 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga 6721 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 6781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 6841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 6901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc 6961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 7021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 7081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 7141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 7201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 7261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 7321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 7381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 7441 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 7501 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 7561 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 7621 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 7681 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca 7741 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg 7801 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg 7861 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca 7921 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg 7981 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt 8041 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact 8101 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg 8161 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta 8221 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag 8281 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt 8341 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt 8401 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg 8461 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg 8521 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat 8581 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg 8641 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt 8701 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc 8761 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg 8821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 8881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag 8941 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg 9001 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat 9061 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg 9121 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag 9181 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat 9241 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt 9301 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc 9361 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg 9421 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac 9481 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat 9541 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc 9601 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat 9661 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag 9721 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc 9781 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac 9841 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg 9901 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa 9961 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca 10021 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg 10081 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc 10141 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg 10201 cgatataggc gccagcaacc gcacctgtgg cg //