LOCUS Exported 10417 bp ds-DNA circular SYN 05-JUL-2019 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10417) AUTHORS ICES TITLE Direct Submission JOURNAL Exported Jul 5, 2019 from SnapGene 4.2.11 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10417 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..302 /label=P6 /label=P6 - BbsI misc_feature 303..3665 /label=sth1Cas9 terminator 3830..3859 /label=fd-ter promoter 3860..4144 /label=P3 misc_feature complement(4174..4523) /label=LacZ cassette misc_feature 4682..4731 /label=Term promoter complement(4759..4855) /label=KasOP misc_feature 4946..5534 /label=ori CDS 5738..6538 /codon_start=1 /label=aac /label=aac(3)IV /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG" CDS complement(7480..9438) /codon_start=1 /label=rep /label=rep (pSG5) - BbsI /translation="MPGSTGPRNGTSPGLASRARRPRNWRSMPSEDVCSVTSVRENSAS TPPRPAGPKCHCGTPIVHIPGKRPKIYCSDACRKRAKRLIADQSRSVASTTTAGNPDAG FSSKGKSPSTAGQSAVTGTGSTPNEGAATAGREDPRDERFARRDRHQTVSLNEAFKGCG TRLTAGTAELMWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLADAGYG LHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGIIGYE RAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFHDALAGAWAAGVESAGGYTVSKTC DRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVSKIGLELAGQNF KAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLNRLIK ELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGLPGVR RLVESWGLVCGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANEARTE ELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKLAPVL QPWGRLPGDCLRVDTAVA" misc_feature 9930..10417 /label=oriT misc_feature 10089 /label=oriT nicking site ORIGIN 1 gccctgcagg cggaagtcag gtagacacga cttccgctag tccttgcaag gtctgctgac 61 gtgaggcggg gcggtcgttt ttgaccgccc tgccttcgtc atgtaggctc gctcgctgtg 121 cctggcgtgt catcagacgc ccaggtcccg gtgccgtgag gcccgggcca tcgagccggt 181 ggtacgtggc tgcggtcccc ttgtgagggc tgcgcgccgt gtgctgtccg gcgcgcacag 241 ccttgaatcc acccgcgggg gccggccggt ctccgtgagc tcgagtagac gacggagacg 301 taatgtcgga cctggtgctg ggcctggaca tcggcatcgg ctccgtcggc gtcggcatcc 361 tgaacaaggt gaccggcgag atcatccaca agaactcccg catcttcccg gccgcacagg 421 ccgagaacaa cctggtccgc cgcaccaacc gccagggccg ccgcctggcc cgccgcaaga 481 agcaccgccg cgtccgcctg aaccgcctgt tcgaggagtc cggcctgatc accgacttca 541 ccaagatcag catcaacctg aacccgtacc agctgcgcgt caagggcctg accgacgagc 601 tgtccaacga ggagctgttc atcgccctga agaacatggt caagcaccgc ggcatctcct 661 acctggacga cgcctccgac gacggcaact cctccgtcgg cgactacgcc cagatcgtca 721 aggagaactc caagcagctg gaaaccaaga cccccggcca gatccagctg gagcgctacc 781 agacctacgg ccagctgcgc ggcgacttca ccgtcgagaa ggacggcaag aagcaccgcc 841 tgatcaacgt gttcccgacc tccgcctacc gctccgaggc cctgcgcatc ctccagaccc 901 agcaggagtt caacccgcag atcaccgacg agttcatcaa ccgctacctg gagatcctga 961 ccggcaagcg caagtactac cacggccccg gcaacgagaa gtcccgcacc gactacggcc 1021 gctaccgcac ctccggcgag acgctggaca acatcttcgg catcctgatc ggcaagtgca 1081 ccttctaccc ggacgagttc cgcgccgcca aggcctccta caccgcccag gagttcaacc 1141 tgctgaacga cctgaacaac ctgaccgtcc cgaccgagac gaagaagctg tccaaggagc 1201 agaagaacca gatcatcaac tacgtcaaga acgagaaggc gatgggcccg gccaagctgt 1261 tcaagtacat cgccaagctg ctgtcctgcg acgtcgccga catcaagggc taccgcatcg 1321 acaagtccgg caaggccgag atccacacct tcgaggccta ccgcaagatg aaaaccctgg 1381 agacgctgga catcgagcag atggaccgcg agacgctgga caagctggcc tacgtcctga 1441 ccctgaacac cgagcgggag ggcatccagg aagccctgga gcacgagttc gccgacggct 1501 ccttctccca gaagcaggtg gacgagctgg tccagttccg caaggccaac tcctccatct 1561 tcggcaaggg ctggcacaac ttctccgtca agctgatgat ggagctgatc ccggagctgt 1621 acgagacgtc cgaggagcag atgaccatcc tgacccgcct gggcaagcaa aagaccacct 1681 cctcctccaa caagaccaag tacatcgacg agaagctgct gaccgaggaa atctacaacc 1741 cggtcgtcgc caagtccgtc cgccaggcca tcaagatcgt caacgccgcc atcaaggagt 1801 acggcgactt cgacaacatc gtcatcgaga tggcccgcga gacgaacgag gacgacgaga 1861 agaaggccat ccagaagatc cagaaggcca acaaggacga gaaggacgcc gccatgctga 1921 aggccgccaa ccagtacaac ggcaaggccg agctgccgca ctccgtgttc cacggccaca 1981 agcagctggc caccaagatc cggctgtggc accagcaggg cgagcgctgc ctgtacaccg 2041 gcaagaccat ctccatccac gacctgatca acaactccaa ccagttcgag gtggaccaca 2101 tcctgccgct gtccatcacc ttcgacgact ccctggccaa caaggtcctg gtctacgcca 2161 ccgccaacca ggagaagggc cagcgcaccc cgtaccaggc cctggactcg atggacgacg 2221 cctggtcctt ccgcgagctg aaggccttcg tccgcgagtc caagaccctg tccaacaaga 2281 agaaggagta cctgctgacc gaagaggaca tctccaagtt cgacgtccgc aagaagttca 2341 tcgagcgcaa cctggtggac acccggtacg cctcccgcgt cgtcctgaac gccctccagg 2401 agcacttccg cgcccacaag atcgacacca aggtgtccgt cgtccgcggc cagttcacct 2461 cccagctccg ccgccactgg ggcatcgaaa agacccgcga cacctaccac caccacgccg 2521 tggacgccct gatcatcgcc gcctcctccc agctgaacct gtggaagaag cagaagaaca 2581 ccctggtgtc ctactccgag gaccagctgc tggacatcga gacgggcgag ctgatctccg 2641 acgacgagta caaggagtcc gtgttcaagg ccccgtacca gcacttcgtg gacaccctga 2701 agtccaagga gttcgaggac tccatcctgt tctcctacca ggtggactcc aagttcaacc 2761 gcaagatcag cgacgccacc atctacgcca cccgccaggc caaggtcggc aaggacaagg 2821 ccgacgagac gtacgtcctg ggcaagatca aggacatcta cacccaggac ggctacgacg 2881 ccttcatgaa aatctacaag aaggacaagt ccaagttcct gatgtaccgc cacgacccgc 2941 agaccttcga gaaggtcatc gagccgatcc tggagaacta cccgaacaag cagatcaacg 3001 acaagggcaa ggaagtcccg tgcaacccgt tcctgaagta caaggaagag cacggctaca 3061 tccgcaagta ctccaagaag ggcaacggcc cggagatcaa gtccctgaag tactacgact 3121 ccaagctggg caaccacatc gacatcaccc cgaaggactc caacaacaag gtcgtcctcc 3181 agtccgtctc cccgtggcgc gccgacgtct acttcaacaa gaccaccggc aagtacgaga 3241 tcctgggcct gaagtacgcc gacctccagt tcgacaaggg caccggcacc tacaagatca 3301 gccaggagaa gtacaacgac atcaagaaga aggaaggcgt ggactccgac tccgagttca 3361 agttcaccct gtacaagaac gacctgctgc tggtcaagga caccgagacg aaggagcagc 3421 agctgttccg cttcctgtcc cgcaccatgc cgaagcagaa gcactacgtc gagctgaagc 3481 cgtacgacaa gcagaagttc gagggcggcg aggccctgat caaggtcctg ggcaacgtcg 3541 ccaactccgg ccagtgcaag aagggcctgg gcaagtccaa catctccatc tacaaggtcc 3601 gcaccgacgt cctgggcaac cagcacatca tcaagaacga gggcgacaag ccgaagctcg 3661 acttccagat ctacgcgttc cccgcaaaag cggcctttga ctccctgcaa gcctcagcga 3721 ccgaatatat cggttatgcg tgggcgatgg ttgttgtcat tgtcggcgca actatcggta 3781 tcaagctgtt taagaaattc acctcgaaag caagctgata aaccgataca attaaaggct 3841 ccttttggag cctttttttg ctgctccttc ggtcggacgt gcgtctacgg gcaccttacc 3901 gcagccgtcg gctgtgcgac acggacggat cgggcgaact ggccgatgct gggagaagcg 3961 cgctgctgta cggcgcgcac cgggtgcgga gcccctcggc gagcggtgtg aaacttctgt 4021 gaatggcctg ttcggttgct ttttttatac ggctgccaga taaggcttgc agcatctggg 4081 cggctaccgc tatgatcggg gcgttcctgc aattcttagt gcgagtatct gaaaggggat 4141 acgcaagtct tctcagccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg 4201 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 4261 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 4321 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc 4381 cctcgaggtc ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat 4441 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4501 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4561 ttgcgttgcg ctcactgccc gctttccacc ggtgaagaca tgtttttgta ctcgaaagaa 4621 gctacaaaga taaggcttca tgccgaaatc aacaccctgt cattttatgg cagggtgttt 4681 tactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt atctagagct 4741 agcatgcata tgaagctttg ttcacattcg aacggtctct gctttgacaa catgctgtgc 4801 ggtgttgtaa agtcgtggcc aggagaatac gacagcgtgc aggactgggg gagttatgac 4861 tagtgtgaaa agtcagtatc cagtcgtgta gttcttatta ccttctagag gccaggaacc 4921 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 4981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 5041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 5161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 5221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 5281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 5341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 5401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 5461 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 5521 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 5581 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 5641 ttttggttca tgtgcagctc catcagcaaa aggggatgat aagtttatca ccaccgacta 5701 tttgcaacag tgccgttgat cgtgctatga tcgactgatg tcatcagcgg tggagtgcaa 5761 tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt cagcttctca accttggggt 5821 tacccccggc ggtgtgctgc tggtccacag ctccttccgt agcgtccggc ccctcgaaga 5881 tgggccactt ggactgatcg aggccctgcg tgctgcgctg ggtccgggag ggacgctcgt 5941 catgccctcg tggtcaggtc tggacgacga gccgttcgat cctgccacgt cgcccgttac 6001 accggacctt ggagttgtct ctgacacatt ctggcgcctg ccaaatgtaa agcgcagcgc 6061 ccatccattt gcctttgcgg cagcggggcc acaggcagag cagatcatct ctgatccatt 6121 gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt gtccatgaac tcgatgggca 6181 ggtacttctc ctcggcgtgg gacacgatgc caacacgacg ctgcatcttg ccgagttgat 6241 ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt cttcaggatg gcaagttggt 6301 acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc tttgccttgg cggacaggtg 6361 gctcaaggag aagagccttc agaaggaagg tccagtcggt catgcctttg ctcggttgat 6421 ccgctcccgc gacattgtgg cgacagccct gggtcaactg ggccgagatc cgttgatctt 6481 cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc gctcgccagt cgattggctg 6541 agctcatgag cggagaacga gatgacgttg gaggggcaag gtcgcgctga ttgctggggc 6601 aacacgtgga gcggatcggg gattgtcttt cttcagctcg ctgatgatat gctgacgctc 6661 aatgccgttt ggcctccgac taacgaaaat cccgcatttg gacggctgat ccgattggca 6721 cggcggacgg cgaatggcgg agcagacgct cgtccggggg caatgagata tgaaaaagcc 6781 tgaactcacc gcgacgtatc gatgtcgagg ttcctcaggg gagccacccc agagaagccc 6841 tcggagctga gcggagctat ttccaaagcc atgccagcta gagacagtgc acaccgccaa 6901 gcatctgcaa aaccctcgcc tggagggaaa gtgcaatgta cgtacgctgg tttccctcca 6961 gaaagggatt ctgcggcttc tcaactttgg aagaagaggc ggaacgaact gctgtttcag 7021 cctactcatg tgaggaggct ggtcctttac cctgatgccc gagggcatca gggtaaagga 7081 ccagcctcgc caactaggag cgctaccaag gcgaacacaa cccaggactt ggtagaaccc 7141 ttcgccggag aagttcagac tcatcggcat gagcacccct cgccgcgcac tcggcaccgg 7201 tccgtcgacc accaccagga cgtcgttgtc gacgtcggcc ccgcggctcc tgcccgccga 7261 acgcgtcgtc gtcgacggcc tggtgctcat cgacgagcac ccggagccag gtgaaaagcg 7321 ccggcggacg ctcggactgg gcgcgggatt ccagcagtaa cccaggtccg ccggccacct 7381 cacggcaggc agaccctcgg ctttcgcgcc gggaccgcca tgagaccgcc acccggatgt 7441 ccggggtggc ggtctcatgg cggtccctca gcggccctac gcgaccgctg tgtcgacgcg 7501 gaggcagtct ccggggagcc ggccccaggg ctggagtacc ggggcgagct tgcccttgca 7561 gcggcggcac acgggcgacg cggcgccggg cgggggagtc ttgacgtcga cggtgaccgg 7621 cttcggcttc aggcgcttcc gaaggctgat cgtcgggcgg atcgcctcct tcttcggctg 7681 acgcactcgg tcgagcgaag cggccagctc ctccgtgcgg gcctcgttgg ccttccggcg 7741 cgcctcgcgg aaagcagctt cctcctcgga catgacctcg aacctcatct ggtcagcgtc 7801 aaggtcgccc ggcgcggccg gcgcttccgg ggcgggcggg tccaggacgt ccttgccaca 7861 caccaggccc caggactcga cgagccgccg gacgcccggt aggccgtacg tctcggcgac 7921 cttgatgagg tcgaggcgac gtccggcgac gcgggcgatg tatcggtacc agatgtaggc 7981 cgggatgacc gcgatggcga ccaggccctc ggtgtcgtcg gtgatctcct cctcggtgcg 8041 gacgtcctgc tggatgccga gttccttgat cagccggttc aggttctgcg accggtagtg 8101 cttgcggacc tggaacacgc cgaactcgcg ctcgcggtac ttctcgacga acgggccggg 8161 ccgacgaagc cgctgcagct cggcggccgc cgcgtcgccc aggtcgagcg gtcccatgcg 8221 gtcgtcgccg cgaccggcct tgaagttctg tccggccagc tccaggccga tcttggagac 8281 gccgcccttg gtcttgtcgc cgtccttgta gaggtagcgg gcctgcttgc ccgcatcgcc 8341 gtcagcggcg tccgcgccgt tgagtgggcg cacgtcggtg ccgtggccct tgccctcaca 8401 ggagcaaccg ggccggtcgc acgtcttgct gacggtgtag ccgcccgcgg attcgacccc 8461 ggcggcccag gctccggcga gtgcgtcgtg gaacgcggcc tgggcgtccg ggccgagcac 8521 ctcgcgggtg acccagagcg tgtgccagtg caggtgccag ccggagcccc agccgaaggt 8581 gtcctcgaag gcccgctcgt agccgatgat cccgaagtcg tcgcgcatcg tgcgccagcg 8641 gcggccggac gagccgtacg cgcccttcca gccgtcgtgc aagaccgcga ccaggccgtg 8701 ccgcattccc ttgcggacgg tgccgaacgc catgcgctcg aagtggcgca acgtgttcgt 8761 gccaaggtgc agcccgtacc cggcgtccgc gagaccgtcg gcggcgagct gcacgttcga 8821 gccccgtacg gccaggatgc ggctcatgca ccacgggcag gtgtggacgt tgttgcagcg 8881 gcacgtgttg ccccacgtcg cctcgcccgg cttccacatc agctcggccg tcccggcagt 8941 gagccgggtc ccgcagccct tgaacgcctc gttcagcgac accgtctggt gccggtcccg 9001 ccgggcgaac cgctcgtcgc gcgggtcctc ccgcccggct gtcgcggcac cctcgtttgg 9061 ggtagaaccc gttccagtta cagcgctctg acctgcagtg gacggagatt ttcccttact 9121 actaaagccc gcgtccggat tacccgctgt agtcgtgctt gctacgctgc gtgactggtc 9181 cgcaatgaga cgctttgcgc gctttcggca ggcgtccgag cagtagattt tggggcgctt 9241 cccggggatg tggacgatcg gggtgccgca gtggcacttc ggtccggcgg ggcgcggtgg 9301 tgtcgacgcg ctgttctctc gtacgctcgt cacagagcaa acgtcctcac tcggcatgct 9361 gcgccagttg cgggggcggc gagcccggga ggccaatccc gggctcgtgc catttctggg 9421 tcctgttgat cctggcattg gtgtggccgt tcattgcccc tgctcgctcc tgacgcgccg 9481 atagacgtcc gatacgcccg gtgctggtgg gatttgatag gtcggaagaa gccccgcccg 9541 ggcctgggcg gggcttcctg tgcgtcagga cctcctcgtc gtgagcctct tcggcctatg 9601 gacggagtga cctcgtgatc cgttacagcc gcgcgcgctc gcgtagagcg gtctcatcag 9661 ttccacgaac ggtcctcttc gcagatcagg gcgttggggc ggagtctcac caaggactac 9721 gtctgctggc gatttccgtt acaccccggg cggtggccgg cgcacacgcg cgcccgcgtt 9781 gggcagtgca gaaagtgcag aaacctaggc gctgatggtc caggtccacg gttcgtcgtc 9841 ggcggcggcg cgggcggcgg cgtcggccag ggcgcgggcg agaccggcta cggcgggctt 9901 gatgcgccgg ttgcgggcga ccttgagcag ctagtatgca ggtcgacgga tcttttccgc 9961 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc 10021 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg 10081 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt 10141 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccggctac 10201 cgccggcgta acagatgagg gcaagcggat ggctgatgaa accaagccaa ccaggaaggg 10261 cagcccacct atcaaggtgt actgccttcc agacgaacga agagcgattg aggaaaaggc 10321 ggcggcggcc ggcatgagcc tgtcggccta cctgctggcc gtcggccagg gctacaaaat 10381 cacgggcgtc gtggactatg agcacgtccg cgagctg //