LOCUS Exported 9318 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expressing SpCas9/VRQR mutant and sgRNA for target sequence with NGA PAM. ACCESSION . VERSION . KEYWORDS pCas9/VRQR-sgRNA (BbsI) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9318) AUTHORS Li S, Prasanna X, Salo VT, Vattulainen I, Ikonen E TITLE An efficient auxin-inducible degron system with low basal degradation in human cells. JOURNAL Nat Methods. 2019 Aug 26. pii: 10.1038/s41592-019-0512-x. doi: 10.1038/s41592-019-0512-x. PUBMED 31451765 REFERENCE 2 (bases 1 to 9318) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9318 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(14..31) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(185..773) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(265..284) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(944..1804) /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 1567..1586 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1805..1909) /gene="bla" /label=AmpR promoter promoter 1962..2202 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1962..1982 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2133..2152 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 2229..2304 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 2462..2841 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2842..3045 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2996..3016 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 3090..3109 /label=T7 /note="T7 promoter, forward primer" promoter 3090..3108 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 3138..3147 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3147..3212 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 3219..3239 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3264..4979 /codon_start=1 /product="N-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)" /label=Cas9(N) /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE" CDS 7365..7412 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 7428..7481 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 7482..8186 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(7624..7642) /label=mCherry-R /note="mCherry, reverse primer" CDS 8202..8258 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 8265..8864 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(8265..8284) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 8761..8781 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(9058..9077) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 9102..9149 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 61 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 1021 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 1081 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 1141 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 1201 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 1261 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 1321 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 1381 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 1441 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 1501 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 1561 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 1621 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 1681 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 1741 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 1801 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 1861 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 1921 gaaaagtgcc acctgacggt accgctagcg tcgacggatc cgagggccta tttcccatga 1981 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga 2041 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt 2101 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa 2161 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gggtcttcga 2221 gaagacctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt 2281 gaaaaagtgg caccgagtcg gtgctttttt tagatctcga gtctagagga ggtcgctgag 2341 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag 2401 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt 2461 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc 2521 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 2581 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 2641 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 2701 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 2761 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 2821 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 2881 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 2941 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 3001 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga 3061 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct 3121 gcccgggatc caccggtgcc accatggact ataaggacca cgacggagac tacaaggatc 3181 atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg 3241 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca 3301 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg 3361 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg 3421 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca 3481 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg 3541 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg 3601 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca 3661 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga 3721 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc 3781 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc 3841 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg 3901 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga 3961 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga 4021 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg 4081 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca 4141 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca 4201 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc 4261 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc 4321 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca 4381 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga 4441 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga 4501 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga 4561 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc 4621 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca 4681 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc 4741 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc 4801 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa 4861 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca 4921 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt 4981 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat 5041 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg 5101 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg 5161 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc 5221 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc 5281 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca 5341 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt 5401 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta 5461 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc 5521 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac 5581 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc 5641 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt 5701 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt 5761 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca 5821 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag 5881 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc 5941 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg 6001 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga 6061 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga 6121 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca 6181 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg 6241 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca 6301 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg 6361 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca 6421 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt 6481 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata 6541 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga 6601 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct 6661 tcgtgagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt 6721 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct 6781 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc 6841 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc 6901 tggcctctgc cagggaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga 6961 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc 7021 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca 7081 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct 7141 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta 7201 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga 7261 agcagtacag gagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg 7321 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca 7381 cgaaaaaggc cggccaggca aaaaagaaaa aggaattcgg cagtggagag ggcagaggaa 7441 gtctgctaac atgcggtgac gtcgaggaga atcctggccc agtgagcaag ggcgaggagg 7501 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga 7561 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 7621 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc 7681 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact 7741 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 7801 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga 7861 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaaa accatgggct 7921 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc 7981 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg 8041 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct 8101 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca 8161 ccggcggcat ggacgagctg tacaaggaat tcggaagcgg cgcaacaaac ttctcactac 8221 tcaaacaagc aggtgacgtg gaggagaatc ccgggcctct taagatgacc gagtacaagc 8281 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg 8341 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg 8401 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg 8461 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg 8521 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc 8581 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg 8641 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc 8701 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc 8761 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg 8821 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaatcgat gcgggactct 8881 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 8941 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 9001 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga 9061 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa 9121 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca 9181 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc 9241 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc 9301 aggccctgcc atagcctc //