LOCUS Exported 6862 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial T. denticola Cas9 (TD) + tracrRNA expression, cloDF13/spectinomycin. ACCESSION . VERSION . KEYWORDS DS-TDcas SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6862) AUTHORS Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM TITLE Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. JOURNAL Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. PUBMED 24076762 REFERENCE 2 (bases 1 to 6862) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6862 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..739 /direction=RIGHT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." promoter 787..878 /gene="bla" /label=AmpR promoter CDS 879..1670 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" terminator 1898..1925 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" repeat_region 6646..6681 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" ORIGIN 1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atcttttgcc ctgtaaacga 61 aaaaaccacc tggggaggtg gtttgatcga aggttaagtc agttggggaa ctgcttaacc 121 gtggtaactg gctttcgcag agcacagcaa ccaaatctgt ccttccagtg tagccggact 181 ttggcgcaca cttcaagagc aaccgcgtgt ttagctaaac aaatcctctg cgaactccca 241 gttaccaatg gctgctgcca gtggcgtttt accgtgcttt tccgggttgg actcaagtga 301 acagttaccg gataaggcgc agcagtcggg ctgaacgggg agttcttgct tacagcccag 361 cttggagcga acgacctaca ccgagccgag ataccagtgt gtgagctatg agaaagcgcc 421 acacttcccg taagggagaa aggcggaaca ggtatccggt aaacggcagg gtcggaacag 481 gagagcgcaa gagggagcga cccgccggaa acggtgggga tctttaagtc ctgtcgggtt 541 tcgcccgtac tgtcagattc atggttgagc ctcacggctc ccacagatgc accggaaaag 601 cgtctgttta tgtgaactct ggcaggaggg cggagcctat ggaaaaacgc caccggcgcg 661 gccctgctgt tttgcctcac atgttagtcc cctgcttatc cacggaatct gtgggtaact 721 ttgtatgtgt ccgcagcgcc cgccgcagtc tcacgcccgg agcgtagcga ccgagtgagc 781 tagctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 841 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagggaagcg gtgatcgccg 901 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt 961 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata 1021 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca 1081 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag 1141 tcaccattgt tgtgcacgac gacatcattc cttggcgtta tccagctaag cgcgaactgc 1201 aatttggaga atggcagcgc aatgacattc ttgcgggtat cttcgagcca gccacgatcg 1261 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc 1321 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg 1381 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc 1441 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg 1501 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta 1561 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag 1621 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa tgtctaacaa 1681 ttcgttcaag ccgaggggcc gcaagatccg gccacgatga cccggtcgtc ggttcagggc 1741 agggtaccag gcacgcctaa ccgtcagtga gattggatga gtgaacgata ttgatcgaga 1801 agagccctgc gcagccgctg ccgtgcctgc aggaagcaac ggcccggagg gtggcgggca 1861 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc 1921 ctttttgcgt ttctacaaac tctgctagct tctagagcac agctaacacc acgtcgtccc 1981 tatctgctgc cctaggtcta tgagtggttg ctggataact ttacgggcat gcataaggct 2041 cgtatgatat attcagggag accacaacgg tttccctcta caaataattt tgtttaactt 2101 ttactagagg aggaggcaaa aatgaagaag gagatcaagg actacttcct gggcctggac 2161 gtgggcaccg gcagcgtggg ctgggccgtg accgacaccg actacaagct gctgaaggcc 2221 aaccgcaagg acctgtgggg catgcgctgc ttcgagactg ctgagaccgc cgaggtgcgc 2281 cggctgcacc gcggtgctcg ccggcgcatc gagcgccgca agaagcgcat caagctgctg 2341 caggagctgt tcagccagga gatcgccaag accgacgagg gcttcttcca gcgcatgaag 2401 gagagcccct tctacgccga ggacaagacc atcctgcagg agaacaccct gttcaacgac 2461 aaggacttcg ccgacaagac ctaccacaag gcctacccca ccatcaacca cctgatcaag 2521 gcctggatcg agaacaaggt gaagcccgac cctcgcctgc tgtacctggc ctgccacaac 2581 atcatcaaga agcgcggcca cttcctgttc gagggcgact tcgacagcga gaaccagttc 2641 gacaccagca tccaggccct gttcgagtac ctgcgcgagg acatggaggt ggacatcgac 2701 gccgacagcc agaaggtgaa ggagatcctg aaggacagca gcctgaagaa cagcgagaag 2761 cagagccgcc tgaacaagat cctgggcctg aagcccagcg acaagcagaa gaaggccatc 2821 accaacctga tcagcggcaa caagatcaac ttcgccgacc tgtacgacaa ccccgacctg 2881 aaggacgccg agaagaacag catcagcttc agcaaggacg acttcgacgc cctgagcgac 2941 gacctggcca gcatcctggg cgacagcttc gagctgctgc tgaaggccaa ggccgtgtac 3001 aactgcagcg tgctgagcaa ggtgatcggc gacgagcagt acctgagctt cgccaaggtg 3061 aagatctacg agaagcacaa gaccgacctt accaagctga agaacgtgat caagaagcac 3121 ttccccaagg actacaagaa ggtgttcggc tacaacaaga acgagaagaa caacaacaac 3181 tacagcggct acgtgggcgt gtgcaagacc aagagcaaga agctgatcat caacaacagc 3241 gtgaaccagg aggacttcta caagttcctg aagaccatcc tgagcgccaa gagcgagatc 3301 aaggaggtga acgacatcct gaccgagatc gagaccggca ccttcctgcc caagcagatc 3361 agcaagagca acgccgagat cccctaccag ctgcgcaaga tggagctgga gaagatcctg 3421 agcaacgccg agaagcactt cagcttcctg aagcagaagg acgagaaggg cctgagccac 3481 agcgagaaga tcatcatgct gctgaccttc aagatcccct actacatcgg ccccatcaac 3541 gacaaccaca agaagttctt ccccgaccgc tgctgggtgg tgaagaagga gaagagcccc 3601 agcggcaaga ccaccccctg gaacttcttc gaccacatcg acaaggagaa gaccgccgag 3661 gccttcatca ccagccgcac caacttctgc acatacctgg tgggcgagag cgtgctgccc 3721 aagagcagcc tgctgtacag cgagtacacc gtgctgaacg agatcaacaa cctgcagatc 3781 atcatcgacg gcaagaacat ctgcgacatc aagctgaagc agaagatcta cgaggacctg 3841 ttcaagaagt acaagaagat cacccagaag cagatcagca ccttcatcaa gcacgatggg 3901 catctgcaac aagaccgacg aggtgatcat cctgggcatc gacaaggagt gcaccagcag 3961 cctgaagagc tacatcgagc tgaagaacat cttcggcaag caggtggacg agatcagcac 4021 caagaacatg ctggaggaga tcatccgctg ggccaccatc tacgacgagg gcgagggcaa 4081 gaccatcctg aagaccaaga tcaaggccga gtacggcaag tactgcagcg acgagcagat 4141 caagaagatc ctgaacctga agttcagcgg ctggggccgc ctgagccgca agttcctgga 4201 gaccgtgacc agcgagatgc ccggcttcag cgagcccgtg aacatcatca ccgccatgcg 4261 cgagacccag aacaacctga tggagctgct gagcagcgag ttcaccttca ccgagaacat 4321 caagaagatc aacagcggct tcgaggacgc cgagaagcag ttcagctacg acggcctggt 4381 gaagcccctg ttcctgagcc ccagcgtgaa gaagatgctg tggcagaccc tgaagctggt 4441 gaaggagatc agccacatca cccaggctcc tcctaagaag atcttcatcg agatggccaa 4501 gggcgccgag ctggagcctg ctcgcaccaa gacccgcctg aagatcctgc aggacctgta 4561 caacaactgc aagaacgacg ccgacgcatt cagcagcgag atcaaggacc tgagcggcaa 4621 gatcgagaac gaggacaacc tgcgcctgcg cagcgacaag ctgtacctgt actacaccca 4681 gctgggcaag tgcatgtact gcggcaagcc catcgagatc ggccacgtgt tcgacaccag 4741 caactacgac atcgaccaca tctaccccca gagcaagatc aaggacgaca gcatcagcaa 4801 ccgcgtgctg gtgtgcagca gctgcaacaa gaacaaggag gacaagtacc ctctgaagag 4861 cgagatccag agcaagcagc gcggcttctg gaacttcctg cagcgcaaca acttcatcag 4921 cctggagaag ctgaaccgcc tgacccgcgc cacccccatc agcgacgacg agaccgccaa 4981 gttcatcgcc cgccagctgg tggagactcg ccaagctacc aaggtggccg ccaaggtgct 5041 ggagaagatg ttccccgaga ccaagatcgt gtacagcaag gccgagaccg tgagcatgtt 5101 ccgcaacaag ttcgacatcg tgaagtgccg cgagatcaac gacttccacc acgcccacga 5161 cgcctacctg aacatcgtgg tgggcaacgt gtacaacacc aagttcacca acaacccctg 5221 gaatttcatt aaggagaagc gcgacaaccc caagatcgcc gacacctaca actactacaa 5281 ggtgttcgac tacgacgtga agcgcaacaa catcaccgcc tgggagaagg gcaagaccat 5341 catcaccgtg aaggacatgc tgaagcgcaa cacccccatc tacacccgcc aggccgcctg 5401 caagaagggc gagctgttca accagaccat catgaagaag ggcctgggcc agcaccccct 5461 gaagaaggag ggccccttca gcaacatcag caagtacggc ggctacaaca aggtgagcgc 5521 cgcctactac accctgatcg agtacgagga gaagggcaac aagatccgca gcctggagac 5581 catccccctg tacctggtga aggacatcca gaaggaccag gacgtgctga agagctacct 5641 gaccgacctg ctgggcaaga aggagttcaa gatcctggtg cccaagatca agatcaacag 5701 cctgctgaag atcaacggct tcccctgcca catcaccggc aagaccaacg acagcttcct 5761 gctgcgcccc gccgtgcagt tctgctgcag caacaacgag gtgctgtact tcaagaagat 5821 catccgcttc agcgagatcc gcagccagcg cgagaagatc ggcaagacca tcagccccta 5881 cgaggacctg agcttccgca gctacatcaa ggagaacctg tggaagaaga ccaagaacga 5941 cgagatcggc gagaaggagt tctacgacct gctgcagaag aagaacctgg agatctacga 6001 catgctgctg accaagcaca aggacaccat ctacaagaag cgccccaaca gcgccaccat 6061 cgacatcctg gtgaagggca aggagaagtt caagagcctg atcatcgaga accagttcga 6121 ggtgatcctg gagatcctga agctgttcag cgccacccgc aacgtgagcg acctgcagca 6181 catcggcggc agcaagtaca gcggcgtggc caagatcggc aacaagatca gcagcctgga 6241 caactgcatc ctgatctacc agagcatcac cggcatcttc gagaagcgca tcgacctgct 6301 gaaggtgtaa gctaagcttg gtgagggata acccgaccgt ccgctgaata aaaaacgccc 6361 ggcggcaacc gagcgttctg ggaccttgac cattccttta aaaagtatta taataacagc 6421 atacacaatt taagatccat cttaaattac acaacgagtt caaataagaa ttcatcaaaa 6481 tcgtcccttt tgggaccgct cattgtggag catcaaggct taacatggtt aagccttttt 6541 tttcatccgc ggcaccgcat cttgggacaa tgaaaacgtt agtcatggcg cgccttgacg 6601 gctagctcag tcctaggtac agtgctagct taattagtct acgaggtttt agagctatgc 6661 tgttttgaat ggtcccaaaa cctgcagaca agcccggccg gcctaaggcg atgccccctc 6721 gacctcgatc agggaggcgt tcaggacgac tcacaaagaa agccgggcaa tgcccggctt 6781 tttccacgcc tcctgggctg acttcaggtg ctacatttga agagataaat tgcactgaaa 6841 tctagagcgg ttcagtagaa aa //