LOCUS Exported 5681 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes MqsR toxin. Gent-R. ACCESSION . VERSION . KEYWORDS pTOX11 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5681) AUTHORS Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK TITLE A New Suite of Allelic Exchange Vectors for the Scarless Modification of Proteobacterial Genomes. JOURNAL Appl Environ Microbiol. 2019 Jun 14. pii: AEM.00990-19. doi: 10.1128/AEM.00990-19. PUBMED 31201277 REFERENCE 2 (bases 1 to 5681) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5681 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 1..28 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 120..206 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS 423..956 /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" RBS 2096..2104 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" primer_bind 2268..2290 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2429..2448 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" oriT 3129..3238 /note="incP origin of transfer" CDS 3271..3642 /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" rep_origin 4260..4648 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" protein_bind 4683..4699 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." terminator 4735..4981 /label=-lambda tL3 terminator /note="transcription terminator tL3 from phage lambda" promoter complement(5361..5479) /label=rhaB promoter /note="promoter of the E. coli rhaBAD operon, conferring tight induction with L-rhamnose and repression with D-glucose in the presence of RhaR and RhaS (Giacalone et al., 2006)" regulatory 5619..5628 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt ttatggcggg 61 cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga 121 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct 181 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggaga 241 ccccacacta ccatcggggg ccatcgatgc aacgggaatt tgaagacaat aactgcctta 301 aaaaaattac gccccgccct gccactcatc gcagtactgt cgaatccatg tgggagttta 361 ttcttgacac agatatttat gatataataa ctgagtaagc ttaacataag gaggaaaaac 421 atatgttacg cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag 481 gtggctcaag tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca 541 tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac 601 atcagccgga ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg 661 ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg 721 agcagccgcg tagtgagatc tatatctatg atctcgcagt ctccggcgag caccggaggc 781 agggcattgc caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt 841 atgtgatcta cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt 901 tgggcatacg ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaaagat 961 ccttggcggc aagaaaacag gacacttggt atacgtgcta gcggccgcgg tccgatcgag 1021 ctcgagacgt cccgggatgc atttaaatta attaacctgg tttaaacctt aagacgtcct 1081 atcctagggc cctgtacacc atgtgcaccg gttcgaagat atcgtacgtg atcagcgaaa 1141 aaaccccgcc gaagcggggt tttttgcgcg attagctcga agctcatgcg gtaccctctc 1201 atccgccaaa acagccaagc ttgcatgcct gcaggtcgac tctagaaggc cttcactgta 1261 agaaaccgtc gcggccttgg cggatgtcgc gagggctcca attgaattcg cgacggaaca 1321 gggtgctgaa atgattgcta tccgaaaagc cacagtggta cgcaatatcg gtcaccgacg 1381 cttccgaatg gcgcagtaag tggcgcgcct tcattaagcg taaacggttt aaatagcgtt 1441 gcggggttaa gccggtctgt tgtttcagtt ggcggtgcag ggtgcgtaag ctcaggctga 1501 actggtcggc aaccgcatcc cagttaactt cgtccgcgaa gtgatcttct aaccacgcta 1561 acagcagatt caggcggctc gccgaattct ctaaattttc ttgcaggctc gacttgcgta 1621 acagcagtaa cagctgcatg aacagaattt cacggctcgc cgtgctcggc aggtcgttct 1681 cgccctcttg ttgctccatt tgcgccacta attggcgaac ttgttgcagc accgaatgat 1741 tcacacgcca atggctaggg tattggccgt ctaattcctg cggtaacagt tggtttaagc 1801 ccgccaggaa ttggaagcgg tcagggctac ggtataaaac gttggttaag cataagttgt 1861 ccgtgtgctc gtataagtgg cggtcgtgat cacgaacgaa gcacacggta ccgcccgtaa 1921 tcgtgtacgg ttggccgttg aaaacgtgga tgccggtacc gtgctccacg ataacgatct 1981 cgtggaagtc gtggtggtgc tccgggaagt cggcttgagg caggcgcggc tcaatggcaa 2041 cgctggcatt gccgctaggg aagaagtcaa ccgagtgcag caccgtcatt aaaaaacctc 2101 cttatcggtg ataagctcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 2161 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgaggc 2221 ctttacctta cttacggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 2281 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcaaggag atggcgccca 2341 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc 2401 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg 2461 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc ctttttgtcc 2521 ggtgttgggt tgaaggtgaa gccggtcggg gccgcagcgg gggccggctt ttcagccttg 2581 cccccctgct tcggccgccg tggctccggc gtcttgggtg ccggcgcggg ttccgcagcc 2641 ttggcctgcg gtgcgggcac atcggcgggc ttggccttga tgtgccgcct ggcgtgcgag 2701 cggaacgtct cgtaggagaa cttgaccttc cccgtttccc gcatgtgctc ccaaatggtg 2761 acgagcgcat agccggacgc taacgccgcc tcgacatccg ccctcaccgc caggaacgca 2821 accgcagcct catcacgccg gcgcttcttg gccgcgcggg attcaaccca ctcggccagc 2881 tcgtcggtgt agctctttgg catcgtctct cgcctgtccc ctcagttcag taatttcctg 2941 catttgcctg tttccagtcg gtagatattc cacaaaacag cagggaagca gcgcttttcc 3001 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc 3061 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg 3121 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact 3181 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct 3241 accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag 3301 ggcagcccac ctatcaaggt gtactgcctt ccagacgaac gaagagcgat tgaggaaaag 3361 gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa 3421 atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg 3481 ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc 3541 ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag agaagcagga cgagcttggc 3601 aaggtcatga tgggcgtggt ccgcccgagg gcagagccat gactttttta gccgctaaaa 3661 cggccggggg gtgcgcgtga ttgccaagca cgtccccatg cgctccatca agaagagcga 3721 cttcgcggag ctggtgaagt acatcaccga cgagcaaggc aagaccgagc gcctgggtca 3781 cgtgcgcgtc acgaactgcg aggcaaacac cctgcccgct gtcatggccg aggtgatggc 3841 gacccagcac ggcaacaccc gttccgaggc cgacaagacc tatcacctgc tggttagctt 3901 ccgcgcggga gagaagcccg acgcggagac gttgcgcgcg attgaggacc gcatctgcgc 3961 tgggcttggc ttcgccgagc atcagcgcgt cagtgccgtg catcacgaca ccgacaacct 4021 gcacatccat atcgccatca acaagattca cccgacccga aacaccatcc atgagccgta 4081 tcgggcctac cgcgccctcg ctgacctctg cgcgacgctc gaacgggact acgggcttga 4141 gcgtgacaat cacgaaacgc ggcagcgcgt ttccgagaac cgcgcgaacg acatggagcg 4201 gcacgcgggc gtggaaagcc tggtcggctg gatccggcca cgatgcgtcc ggcgtagagg 4261 atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta 4321 ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct 4381 aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta ctaagctctc 4441 atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt 4501 tttataagaa aaaaaagaat atataaggct tttaaagctt ttaaggttta acggttgtgg 4561 acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa gcaattttga 4621 gtgacacagg aacacttaac ggctgacatg ggaattccac atgtggaatt ccacatgtgg 4681 aattgtgagc ggataacaat ttgtggaatt cccccacgac ttcttcgtct gtttctactg 4741 gtattggcac aaacctgatt ccaatttgag caaggctatg tgccatctcg atactcgttc 4801 ttaactcaac agaagatgct ttgtgcatac agcccctcgt ttattattta tctcctcagc 4861 cagccgctgt gctttcagtg gatttcggat aacagaaagg ccgggaaata cccagcctcg 4921 ctttgtaacg gagtagagac gaaagtgatt gcgcctaccc ggatattatc gtgaggatgc 4981 gtcatcgcca ttaattcact gatcagtgat aagctgtcaa acatgagaat tcgcggatcc 5041 ttacttctcc ttaaacgaga cgatcagtac gtcatgaatt accgtaattt taagataaac 5101 ctggcctgta acaagcctgg gtctgtaaac atcctgccag atagtatgat cagagtaggt 5161 ggtcatgctt ttataaaagt cgctctctga taatccaatg ataacattac acataccatc 5221 aaaatccaaa cctaactcat ctgcatttaa cagggcacta cgtgttgtac gaacttgccc 5281 ggcattgaca agttttttaa cctgactcaa acgtgtatgt ggtgtgcgtt tttccatatg 5341 gtgatcctgc tgaatttcat tacgaccagt ctaaaaagcg cctgaattcg cgaccttctc 5401 gttactgaca ggaaaatggg ccattggcaa ccagggaaag atgaacgtga tgatgttcac 5461 aatttgctga attgtggtga tgtgatgctc accgcatttc ctgaaaattc acgctgtatc 5521 ttgaaaaatc gacgtttttt acgtggtttt ccgtcgaaaa tttaaggtaa gaacctgacc 5581 tcgtgattac tatttcgccg tgttgacgac atcaggaggc caccatggcc tcgactagtt 5641 ccggacagct gcaggcatgc aagcttggca ctggccacgc a //