LOCUS Exported 9152 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION dCas9 + PA4-mVenus. ACCESSION . VERSION . KEYWORDS pJK384.4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9152) AUTHORS Kuo J, Yuan R, Sanchez C, Paulsson J, Silver PA TITLE Toward a translationally independent RNA-based synthetic oscillator using deactivated CRISPR-Cas. JOURNAL Nucleic Acids Res. 2020 Aug 20;48(14):8165-8177. doi: 10.1093/nar/gkaa557. PUBMED 32609820 REFERENCE 2 (bases 1 to 9152) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9152 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(7..630) /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 538..557 /label=Tet-R /note="Tetracycline resistance gene, reverse primer" promoter 649..704 /gene="tetR" /label=tetR/tetA promoters /note="overlapping promoters for bacterial tetR and tetA" protein_bind 685..703 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" RBS 722..733 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 740..4846 /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" terminator 4870..4941 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4957..4984 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5515..6465) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 6513..6735 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" misc_RNA 7280..7329 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" RBS 7359..7370 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" primer_bind complement(7409..7437) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 8032..8052 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 8153..8239 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 8252..8346 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(8367..9026) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 8941..8960 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(9027..9129) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc 61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg 121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg 181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa 241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc 301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag 361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc 421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa 481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc 541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt 601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta 661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc 721 taaagaggag aaaggatcta tggataagaa atactcaata ggcttagcta tcggcacaaa 781 tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt 841 tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga 901 cagtggagag acagcggaag cgactcgtct caaacggaca gctcgtagaa ggtatacacg 961 tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga 1021 tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga 1081 acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac 1141 tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat 1201 ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt 1261 aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca 1321 attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc 1381 acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa 1441 aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc 1501 aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga 1561 tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa 1621 gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa 1681 ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct 1741 tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca 1801 atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag aattttataa 1861 atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa 1921 tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat 1981 tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa 2041 agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc 2101 attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac 2161 cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg 2221 catgacaaac tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct 2281 ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat 2341 gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa 2401 aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg 2461 ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta 2521 ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga 2581 tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga 2641 aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg 2701 ccgttatact ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca 2761 atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat 2821 gcagctgatc catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc 2881 tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa 2941 aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca 3001 taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca 3061 gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca 3121 gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta 3181 ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag 3241 tgattatgat gtcgatgcca ttgttccaca aagtttcctt aaagacgatt caatagacaa 3301 taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga 3361 agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca 3421 acgtaagttt gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc 3481 tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat 3541 tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa 3601 agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa 3661 agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg 3721 aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa 3781 agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc 3841 aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa 3901 tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg 3961 ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat 4021 tgtcaagaaa acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag 4081 aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt 4141 tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc 4201 gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt 4261 tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt 4321 aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct 4381 ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa 4441 ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca 4501 aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag 4561 tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata 4621 taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc atttatttac 4681 gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa 4741 acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg 4801 tctttatgaa acacgcattg atttgagtca gctaggaggt gactaactcg agtaaggatc 4861 tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt 4921 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc 4981 tgcgtttata cctagggtac gggttttgct gcccgcaaac gggctgttct ggtgttgcta 5041 gtttgttatc agaatcgcag atccggcttc agccggtttg ccggctgaaa gcgctatttc 5101 ttccagaatt gccatgattt tttccccacg ggaggcgtca ctggctcccg tgttgtcggc 5161 agctttgatt cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct 5221 gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca 5281 cctgttctat taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga 5341 acagctttga atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg 5401 ttttcatctg tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta 5461 cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat 5521 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga 5581 gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact 5641 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag 5701 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt 5761 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc 5821 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt 5881 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag 5941 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt 6001 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga 6061 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat 6121 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg 6181 gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc 6241 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc 6301 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg 6361 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg 6421 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc 6481 taggtgattt taatcactat accaattgag atgggctagt caatgataat tactagtcct 6541 tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa 6601 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca 6661 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag 6721 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac 6781 gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc 6841 aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct 6901 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat 6961 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc 7021 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt 7081 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt 7141 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta 7201 cccgtcttac tgtccctagt gcttggccta gtatgcgtct cctttacacc tccacaacta 7261 gctggtatta tgctagcagc tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg 7321 acgaaacagc ctctacaaat aattttgttt aactttacaa agaggagaaa ggtaccgcat 7381 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga 7441 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa 7501 acttaccctt aaattgattt gcactactgg aaaactacct gttccatggc caacacttgt 7561 cactactttg ggttatggtg ttcaatgctt tgcgagatac ccagatcata tgaaacagca 7621 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa 7681 agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa 7741 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt 7801 ggaatacaac tataactcac acaatgtata catcacggca gacaaacaaa agaatggaat 7861 caaagcgaac ttcaaaatta gacacaacat tgaagatgga ggtgttcaac tagcagacca 7921 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct 7981 gtcctaccaa tctaagcttt cgaaagatcc caacgaaaag agagaccaca tggtccttct 8041 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aataagcttg 8101 atatcgaatt cctgcagccc gggggatccc attgtacgcg tgctagaggc atcaaataaa 8161 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc 8221 tctcctgagt aggacaaatt ctagtgcttg gattctcacc aataaaaaac gcccggcggc 8281 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca 8341 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt 8401 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga 8461 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg 8521 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag 8581 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt 8641 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg 8701 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg 8761 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga 8821 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc 8881 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga 8941 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg 9001 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac 9061 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg 9121 tgccgatcaa cgtctcattt tcgccagata tc //