LOCUS Exported 9949 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of 6His-dCas9-3XFlag. ACCESSION . VERSION . KEYWORDS pCT310 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9949) AUTHORS Tsui C, Inouye C, Levy M, Lu A, Florens L, Washburn MP, Tjian R TITLE dCas9-targeted locus-specific protein isolation method identifies histone gene regulators. JOURNAL Proc Natl Acad Sci U S A. 2018 Mar 20;115(12):E2734-E2741. doi: 10.1073/pnas.1718844115. Epub 2018 Mar 5. PUBMED 29507191 REFERENCE 2 (bases 1 to 9949) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9949 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 20..39 /label=T7 /note="T7 promoter, forward primer" promoter 20..38 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 39..63 /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)." CDS 112..129 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 139..162 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 172..4272 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 4297..4362 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" primer_bind complement(4460..4478) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4474..4521 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(4759..4777) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4845..4949 /gene="bla" /label=AmpR promoter CDS 4950..5810 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5168..5187) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5981..6569 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6470..6489 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6723..6740 /label=L4440 /note="L4440 vector, forward primer" misc_feature 6755..6894 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(6820..6839) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6980..7002 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(6996..7187) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(8499..9581) /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 9543..9562 /label=LacI-R /note="LacI, reverse primer" promoter complement(9582..9659) /gene="lacI" /label=lacI promoter primer_bind 9865..9884 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 61 tcccctctag aaataatttt gtttaaactt taagaaggag atatacatat gcatcatcat 121 catcatcacg tgggaggtct ggaagttctg ttccaggggc ccgcagcctc ggataagaaa 181 tactcaatag gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa 241 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa 301 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc 361 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag 421 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct 481 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat 541 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat 601 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt 661 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta 721 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt 781 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat 841 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg 901 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta 961 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat 1021 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat 1081 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc 1141 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca 1201 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat 1261 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat 1321 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc 1381 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga 1441 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg 1501 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg 1561 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa 1621 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat 1681 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg 1741 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag 1801 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta 1861 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa 1921 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa 1981 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc 2041 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat 2101 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga 2161 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa 2221 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt 2281 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt 2341 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt 2401 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca 2461 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc 2521 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact 2581 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg 2641 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa 2701 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt 2761 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga 2821 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa 2881 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact 2941 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat 3001 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct 3061 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc 3121 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt 3181 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag 3241 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac 3301 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa 3361 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc 3421 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga 3481 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa 3541 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta 3601 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta 3661 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct 3721 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt 3781 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat 3841 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag 3901 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat 3961 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat 4021 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa 4081 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt 4141 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat 4201 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag 4261 ctaggaggtg acggtggctc cagatctgcg gccgcggact acaaagacca tgacggtgat 4321 tataaagatc atgacatcga ttacaaggat gacgatgaca agtagggccg ctgactcgag 4381 atcgatgata ttcgagccta ggtataatcg gatccggctg ctaacaaagc ccgaaaggaa 4441 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa 4501 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc 4561 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga 4621 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact 4681 gtgataaact accgcattaa agctagctta tcgatgataa gctgtcaaac atgagaatta 4741 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 4801 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 4861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 4921 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 4981 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 5041 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 5101 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 5161 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc 5221 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 5281 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 5341 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 5401 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 5461 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac 5521 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 5581 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 5641 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 5701 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 5761 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 5821 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 5881 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 5941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 6001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 6061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 6121 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 6181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 6241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 6301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 6361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 6421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 6481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 6541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 6601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 6661 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 6721 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc 6781 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc 6841 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 6901 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 6961 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 7021 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg 7081 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt 7141 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg 7201 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg 7261 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg 7321 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt 7381 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac 7441 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag 7501 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc 7561 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc 7621 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg 7681 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt 7741 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg 7801 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac 7861 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc 7921 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta 7981 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc 8041 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa 8101 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg 8161 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga 8221 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg 8281 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc 8341 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg 8401 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg 8461 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg 8521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 8581 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc 8641 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg 8701 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg 8761 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc 8821 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca 8881 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc 8941 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc 9001 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc 9061 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt 9121 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca 9181 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga 9241 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc 9301 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg 9361 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt 9421 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc 9481 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca 9541 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga 9601 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc 9661 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt 9721 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag 9781 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa 9841 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc 9901 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgac //