LOCUS Exported 5866 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes MqsR toxin. CAM-R. ACCESSION . VERSION . KEYWORDS pTOX2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5866) 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 5866) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5866 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..119 /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)" terminator 499..745 /label=-lambda tL3 terminator /note="transcription terminator tL3 from phage lambda" protein_bind 781..797 /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)." rep_origin 832..1220 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" CDS complement(1838..2209) /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" oriT 2242..2351 /note="incP origin of transfer" primer_bind complement(3032..3051) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(3190..3212) /label=pGEX 3' /note="pGEX vectors, reverse primer" RBS 3376..3384 /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)" promoter 4596..4698 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 4699..5358 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(4765..4784) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 5459..5545 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5637..5664 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" regulatory 5718..5727 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc 61 ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga ctggtcgtaa 121 tgaaattcag caggatcacc atatggaaaa acgcacacca catacacgtt tgagtcaggt 181 taaaaaactt gtcaatgccg ggcaagttcg tacaacacgt agtgccctgt taaatgcaga 241 tgagttaggt ttggattttg atggtatgtg taatgttatc attggattat cagagagcga 301 cttttataaa agcatgacca cctactctga tcatactatc tggcaggatg tttacagacc 361 caggcttgtt acaggccagg tttatcttaa aattacggta attcatgacg tactgatcgt 421 ctcgtttaag gagaagtaag gatccgcgaa ttctcatgtt tgacagctta tcactgatca 481 gtgaattaat ggcgatgacg catcctcacg ataatatccg ggtaggcgca atcactttcg 541 tctctactcc gttacaaagc gaggctgggt atttcccggc ctttctgtta tccgaaatcc 601 actgaaagca cagcggctgg ctgaggagat aaataataaa cgaggggctg tatgcacaaa 661 gcatcttctg ttgagttaag aacgagtatc gagatggcac atagccttgc tcaaattgga 721 atcaggtttg tgccaatacc agtagaaaca gacgaagaag tcgtggggga attccacaaa 781 ttgttatccg ctcacaattc cacatgtgga attccacatg tggaattccc atgtcagccg 841 ttaagtgttc ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt 901 tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa gccttatata 961 ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt 1021 aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc 1081 catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac 1141 gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt 1201 gaactgctga tcttcagatc ctctacgccg gacgcatcgt ggccggatcc agccgaccag 1261 gctttccacg cccgcgtgcc gctccatgtc gttcgcgcgg ttctcggaaa cgcgctgccg 1321 cgtttcgtga ttgtcacgct caagcccgta gtcccgttcg agcgtcgcgc agaggtcagc 1381 gagggcgcgg taggcccgat acggctcatg gatggtgttt cgggtcgggt gaatcttgtt 1441 gatggcgata tggatgtgca ggttgtcggt gtcgtgatgc acggcactga cgcgctgatg 1501 ctcggcgaag ccaagcccag cgcagatgcg gtcctcaatc gcgcgcaacg tctccgcgtc 1561 gggcttctct cccgcgcgga agctaaccag caggtgatag gtcttgtcgg cctcggaacg 1621 ggtgttgccg tgctgggtcg ccatcacctc ggccatgaca gcgggcaggg tgtttgcctc 1681 gcagttcgtg acgcgcacgt gacccaggcg ctcggtcttg ccttgctcgt cggtgatgta 1741 cttcaccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt gcttggcaat 1801 cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc ctcgggcgga 1861 ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg 1921 cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca 1981 gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat 2041 agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag gccgacaggc 2101 tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca 2161 ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct 2221 catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg 2281 ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac 2341 ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc 2401 aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc 2461 cgaagcaggg ttatgcagcg gaaaagcgct gcttccctgc tgttttgtgg aatatctacc 2521 gactggaaac aggcaaatgc aggaaattac tgaactgagg ggacaggcga gagacgatgc 2581 caaagagcta caccgacgag ctggccgagt gggttgaatc ccgcgcggcc aagaagcgcc 2641 ggcgtgatga ggctgcggtt gcgttcctgg cggtgagggc ggatgtcgag gcggcgttag 2701 cgtccggcta tgcgctcgtc accatttggg agcacatgcg ggaaacgggg aaggtcaagt 2761 tctcctacga gacgttccgc tcgcacgcca ggcggcacat caaggccaag cccgccgatg 2821 tgcccgcacc gcaggccaag gctgcggaac ccgcgccggc acccaagacg ccggagccac 2881 ggcggccgaa gcaggggggc aaggctgaaa agccggcccc cgctgcggcc ccgaccggct 2941 tcaccttcaa cccaacaccg gacaaaaagg atcctctacg ccggacgcat cgtggccggc 3001 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa 3061 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc 3121 cccgtggccg ggggactgtt gggcgccatc tccttgctgc ctcgcgcgtt tcggtgatga 3181 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 3241 tgccgtaagt aaggtaaagg cctcgcggaa cccctatttg tttatttttc taaatacatt 3301 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 3361 ggagcttatc accgataagg aggtttttta atgacggtgc tgcactcggt tgacttcttc 3421 cctagcggca atgccagcgt tgccattgag ccgcgcctgc ctcaagccga cttcccggag 3481 caccaccacg acttccacga gatcgttatc gtggagcacg gtaccggcat ccacgttttc 3541 aacggccaac cgtacacgat tacgggcggt accgtgtgct tcgttcgtga tcacgaccgc 3601 cacttatacg agcacacgga caacttatgc ttaaccaacg ttttataccg tagccctgac 3661 cgcttccaat tcctggcggg cttaaaccaa ctgttaccgc aggaattaga cggccaatac 3721 cctagccatt ggcgtgtgaa tcattcggtg ctgcaacaag ttcgccaatt agtggcgcaa 3781 atggagcaac aagagggcga gaacgacctg ccgagcacgg cgagccgtga aattctgttc 3841 atgcagctgt tactgctgtt acgcaagtcg agcctgcaag aaaatttaga gaattcggcg 3901 agccgcctga atctgctgtt agcgtggtta gaagatcact tcgcggacga agttaactgg 3961 gatgcggttg ccgaccagtt cagcctgagc ttacgcaccc tgcaccgcca actgaaacaa 4021 cagaccggct taaccccgca acgctattta aaccgtttac gcttaatgaa ggcgcgccac 4081 ttactgcgcc attcggaagc gtcggtgacc gatattgcgt accactgtgg cttttcggat 4141 agcaatcatt tcagcaccct gttccgtcgc gaattcaatt ggagccctcg cgacatccgc 4201 caaggccgcg acggtttctt acagtgaagg ccttctagag tcgacctgca ggcatgcaag 4261 cttggctgtt ttggcggatg agagggtacc gcatgagctt cgagctaatc gcgcaaaaaa 4321 ccccgcttcg gcggggtttt ttcgctgatc acgtacgata tcttcgaacc ggtgcacatg 4381 gtgtacaggg ccctaggata ggacgtctta aggtttaaac caggttaatt aatttaaatg 4441 catcccggga cgtctcgagc tcgatcggac cgcggccgct agcacgtata ccaagtgtcc 4501 tgttttcttg ccgccaagga tctgatggcg caggggatca agatctctgt tgataccggg 4561 aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac 4621 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca 4681 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc 4741 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac 4801 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag 4861 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt 4921 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt 4981 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg 5041 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc 5101 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc 5161 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc 5221 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc 5281 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag 5341 tggcagggcg gggcgtaatt tttttaaggc agttattgtc ttcaaattcc cgttgcatcg 5401 atggcccccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca 5461 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt 5521 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg 5581 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa 5641 ggccatcctg acggatggcc tttttgcgtg gccagtgcca agcttgcatg cctgcagctg 5701 tccggaacta gtcgaggcca tggtggcctc ctgatgtcgt caacacggcg aaatagtaat 5761 cacgaggtca ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa acgtcgattt 5821 ttcaagatac agcgtgaatt ttcaggaaat gcggtgagca tcacat //