LOCUS Exported 7257 bp ds-DNA circular SYN 10-MAY-2017 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS 89240_sw SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7257) AUTHORS . TITLE Direct Submission JOURNAL Exported Wednesday, May 10, 2017 from SnapGene 3.3.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7257 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 16..35 /label=Sequencing Primer 118Cas1 promoter 27..45 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 65..1170 /label=Strep II cas1 /label=his6 cas1 RBS 77..99 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 119..142 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 143..169 /label=TEV site CDS 143..163 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" primer_bind 1190..1206 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 1255..4588 /label=untagged cas3 RBS 1283..1305 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" terminator 4746..4793 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(4928..5719) /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" rep_origin 5841..6429 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 6615..6754 /label=bom /note="basis of mobility region from pBR322" CDS complement(6856..7047) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 ggcaccatgc atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt 61 tccctctaga aataattttg tttaacttta agaaggagat atacatatga aatcttcttg 121 gagccacccg cagttcgaaa aggaaaacct gtacttccaa tccaatgcag acgacatttc 181 tcccagcgaa ctgaagacga tccttcactc caagcgtgcc aacctgtact acctgcaaca 241 ctgccgggta ctggtcaacg gcgggcgggt cgagtacgtc accgacgaag gccggcattc 301 gcactactgg aacatcccca tcgccaacac caccagcctg ttgctgggca ccggtacctc 361 cattacccag gcagccatgc gcgaactggc cagggccggg gtattggtag gcttctgcgg 421 cggcggcgga acgccgctgt tcagcgccaa cgaagtggac gtcgaggtgt cctggctgac 481 gccgcagagc gaataccgcc ccaccgaata cctgcagcgc tgggtcggct tctggttcga 541 tgaggaaaaa cgcctggtgg ccgcccggca cttccagcga gccaggctgg agcggattcg 601 ccacagttgg ctggaagacc gcgtattgcg cgacgccggc ttcgccgtcg atgccaccgc 661 actggcagtt gccgtggaag attccgcccg agcgttggag caggctccca atcatgagca 721 tctactgacc gaagaagcgc gcctgagcaa gcgcctgttc aagctcgcag cccaggccac 781 ccgctacggc gagttcgtcc gcgccaagcg cggcagcggc ggcgacccgg ccaaccgctt 841 cctcgaccat ggcaactacc tggcctacgg cctcgcggcg accgccacct gggtgctggg 901 cattccccat ggactggcgg tgctacacgg caagacccgg cgcggaggcc tggtgttcga 961 tgtggcggac ctgatcaagg actcgctgat cctgccgcag gcatttcttt ccgccatgcg 1021 cggcgacgag gaacaggact tccgccaggc ctgcctggac aacctgagcc gcgcacaggc 1081 cctggacttc atgatcgaca cgctgaaaga cgtggcgcag cggagcacgg tgtccgcatg 1141 ataataacat tggaagtgga taacggatcc gggccctgta caagatcctg taaaacgacg 1201 gccagtgaat tcactcgagt cccggggcga tcgcagcgga taacaatttc acatcctgca 1261 ggactcgagt tctagaaata attttgttta actttaagaa ggagatatag atcatgaaca 1321 tcctgctggt gtcgcaatgc gaaaagcgcg ccctgagcga aacccgccgc attctcgacc 1381 agttcgccga gcgccgcggc gaacggacct ggcaaacgcc catcactcaa gccggactgg 1441 ataccctgcg acgcctgctg aagaaaagcg cacggcgcaa caccgccgta gcctgtcact 1501 ggatccgcgg ccgcgaccac agcgaactgc tgtggatcgt cggtgatgcc agccgcttca 1561 acgcccaggg tgcggtgccg accaacagga cctgccgcga catcctgcgc aaggaagacg 1621 agaacgactg gcacagcgcc gaggacatcc gcctgctgac ggtgatggca gcgctgttcc 1681 acgatatcgg caaggccagc caggccttcc aggccaaact gcggaaccgc ggcaaaccga 1741 tggccgatgc ctatcgtcac gaatgggtat cactgcgcct gttcgaagcc ttcgttggcc 1801 caggcagcag cgacgaggac tggctgaggc gcctggcgga caagcgagag acgggcgatg 1861 cctggctgtc gcaactggcc agggacgacc ggcaatccgc gccacccggc ccgttccaga 1921 aaagccggct accgccgctc gcccaggcgg tcggctggtt gatcgtcagc catcatcgcc 1981 tgcccaacgg ggaccatcgc ggcagcgcct cgctggcacg cttgccggcc cccatccaga 2041 gccaatggtg cggcgcacgc gacgcagacg caaaagaaaa ggccgcctgc tggcagttcc 2101 cccacggcct gcccttcgcc agcgcccatt ggcgcgccag gacagcgcta tgcgcgcaga 2161 gcatgctcga gcgtcccggc ctgctggctc ggggaccggc cttgttgcat gattcctacg 2221 tcatgcatgt gtcccgactg atcctgatgc tcgcggacca ccactattcc agtctccctg 2281 ccgactcccg gctgggcgac ccgaacttcc ccttgcacgc caacaccgac cgggacagcg 2341 gcaaactaaa gcagcgcctg gacgaacacc tgctcggcgt cgccctgcac agtcgcaagc 2401 tcgccggcac cctgccacgc ctggagcgac aactaccgcg ccttgcccgg cacaagggct 2461 tcacccgccg ggtcgagcag ccgcgcttcc gctggcagga caaggcctac gactgcgcga 2521 tggcctgccg cgagcaggct atggagcatg gattcttcgg cctcaacctg gcgtcgaccg 2581 gttgcggtaa gaccctcgcc aacggccgta tcctgtatgc gctggccgat ccgcaacgcg 2641 gcgcgcgttt cagcatcgct ctcggcctgc gcagcttgac cctgcaaacc gggcaggcct 2701 accgcgagcg gctcggcctg ggcgacgacg acctcgctat cctggtcggc ggcagcgccg 2761 cccgcgaact gttcgaaaag cagcaggagc gcctggagcg cagcggtagc gagtcagccc 2821 aggagctgct ggcggaaaac agccatgtac acttcgccgg cacgctcgag gacggccctc 2881 tacgcgagtg gctcggcagg aacagcgcgg gaaaccgcct actccaggcg cccatcctgg 2941 cctgcaccat cgaccacctg atgcccgcca gcgaaagcct gcgcggcgga caccagatag 3001 cgccactgct ccgcctgatg acttccgacc tggtgctcga cgaggtcgac gacttcgata 3061 tcgacgacct gcccgccctg tcgcggctgg tgcactgggc cggcctgttc ggcagccgcg 3121 tgctgctctc ctccgcgacc ctgccgccgg ccttggtgca gggcctgttc gaggcctatc 3181 gcagcggccg ggaaatcttc cagcgccatc gtggcgctcc cggacgcgct acggaaatcc 3241 gctgtgcctg gttcgacgag ttctccagcc aatccagcgc ccacggcgcc gtaacctcct 3301 tcagcgaagc gcatgcgacc ttcgtcgccc agcgtctggc gaagctcgag caactgccgc 3361 cacgtcgcca ggcgcagcta tgcaccgtgc atgccgctgg cgaggcccgt cccgcgctgt 3421 gccgcgagtt ggccgggcag atgaatacct ggatggctga cctgcatcgc tgccatcaca 3481 ccgaacacca aggacgtcgc atcagtttcg gcctgctacg gctggccaac atcgaacccc 3541 tgatcgaact ggcccaggcc atcctcgccc agggtgcgcc cgaggggttg catgtccatc 3601 tgtgtgtcta ccattcgcgg catccccttc tggtccgctc ggccatcgag cgacaactcg 3661 atgaactgct gaagcgttcg gacgacgacg ccgccgcgct gttcgctcgt ccgacgctgg 3721 ccaaggcgct ccaggccagc acggagcggg atcatctgtt cgtcgtactc gcctcgccgg 3781 tggcggaggt cggtcgcgac cacgattacg actgggccat cgtcgaaccc tcctccatgc 3841 gctcgatcat ccagttggcc gggcgaatcc gccgccatcg ctccggcttc agcggcgagg 3901 ccaacctata cctgctatcg cgcaatatcc gctcgctgga agggcagaat ccggcgttcc 3961 agcggcccgg cttcgagacc cccgacttcc ctcttgacag ccacgacctg cacgacctgc 4021 tcgaccccgc cctactcgcc cgcatcgacg ccagcccacg aatcgtcgaa ccgttcccac 4081 tgttcccacg cagccggttg gtcgacctgg aacaccgacg cctgcgcgcg ctgatgcttg 4141 ccgacgaccc accgtcgtcc ctgctcggcg taccgctctg gtggcaaacc ccggcatcgc 4201 tcagcggcgc cctgcaaacc agccaaccat ttcgcgcagg cgccaaggag cgatgctacg 4261 ccctgctgcc ggacgaggac gacgaggagc gcttgcattt cagccgctac gaagaaggga 4321 cctggagcaa ccaggacaac ctgttgcgca acctcgacct cacctatggc ccgcgcatcc 4381 agacatgggg cacggtcaac tatcgggagg agctagtcgc aatggccggc cgcgaggacc 4441 tcgacctgcg tcaatgcgcc atgcgctacg gcgaggtgag attgcgagaa aacacccagg 4501 gatggagcta ccacccttat ttggggttca agaaatacaa ctgataataa gatcccaact 4561 ccataaggat ccgcgatcgc ggcgcgcccg ccagggtttt cccagtcgcg gccgctcgcg 4621 cacctggtgc catcgccctg atagacgtta attaaagtac tggccggcca cgcgtctcga 4681 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc 4741 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag 4801 gaggaactat atccgactag taacgtttac aatttcaggt ggcacttttc ggggaaatgt 4861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa 4921 ttaattctta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt 4981 ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag 5041 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga 5101 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca 5161 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat 5221 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac 5281 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga 5341 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca 5401 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt 5461 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga 5521 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat 5581 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg 5641 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg 5701 cgatcaccgc ttccctcata acaccccttg tattactgtt tatgtaagca gacagtttta 5761 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 5821 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 5881 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 5941 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 6001 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 6061 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 6121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 6181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 6241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 6301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 6361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 6421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 6481 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 6541 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 6601 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 6661 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac 6721 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 6781 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 6841 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 6901 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc 6961 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 7021 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 7081 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa 7141 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac 7201 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgtt //