LOCUS Exported 6640 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes MqsR toxin and amilCP chromoprotein. CAM-R. ACCESSION . VERSION . KEYWORDS pTOX5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6640) 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 6640) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6640 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..389 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" CDS complement(1007..1378) /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" oriT 1411..1520 /note="incP origin of transfer" primer_bind complement(2201..2220) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(2359..2381) /label=pGEX 3' /note="pGEX vectors, reverse primer" RBS 2545..2553 /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 3431..3459 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" promoter 4539..4641 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 4642..5301 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(4708..4727) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 5402..5488 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5580..5607 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" regulatory 5661..5670 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter 5810..5928 /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 6308..6554 /label=-lambda tL3 terminator /note="transcription terminator tL3 from phage lambda" protein_bind 6590..6606 /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)." ORIGIN 1 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc 61 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag 121 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg 181 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag 241 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga 301 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat 361 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcca 421 gccgaccagg ctttccacgc ccgcgtgccg ctccatgtcg ttcgcgcggt tctcggaaac 481 gcgctgccgc gtttcgtgat tgtcacgctc aagcccgtag tcccgttcga gcgtcgcgca 541 gaggtcagcg agggcgcggt aggcccgata cggctcatgg atggtgtttc gggtcgggtg 601 aatcttgttg atggcgatat ggatgtgcag gttgtcggtg tcgtgatgca cggcactgac 661 gcgctgatgc tcggcgaagc caagcccagc gcagatgcgg tcctcaatcg cgcgcaacgt 721 ctccgcgtcg ggcttctctc ccgcgcggaa gctaaccagc aggtgatagg tcttgtcggc 781 ctcggaacgg gtgttgccgt gctgggtcgc catcacctcg gccatgacag cgggcagggt 841 gtttgcctcg cagttcgtga cgcgcacgtg acccaggcgc tcggtcttgc cttgctcgtc 901 ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca tggggacgtg 961 cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat ggctctgccc 1021 tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct tcgatcttcg 1081 ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc 1141 agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc agctcgcgga 1201 cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc agcaggtagg 1261 ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa 1321 ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc 1381 gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt 1441 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc 1501 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa 1561 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta 1621 taatgacccc gaagcagggt tatgcagcgg aaaagcgctg cttccctgct gttttgtgga 1681 atatctaccg actggaaaca ggcaaatgca ggaaattact gaactgaggg gacaggcgag 1741 agacgatgcc aaagagctac accgacgagc tggccgagtg ggttgaatcc cgcgcggcca 1801 agaagcgccg gcgtgatgag gctgcggttg cgttcctggc ggtgagggcg gatgtcgagg 1861 cggcgttagc gtccggctat gcgctcgtca ccatttggga gcacatgcgg gaaacgggga 1921 aggtcaagtt ctcctacgag acgttccgct cgcacgccag gcggcacatc aaggccaagc 1981 ccgccgatgt gcccgcaccg caggccaagg ctgcggaacc cgcgccggca cccaagacgc 2041 cggagccacg gcggccgaag caggggggca aggctgaaaa gccggccccc gctgcggccc 2101 cgaccggctt caccttcaac ccaacaccgg acaaaaagga tcctctacgc cggacgcatc 2161 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc 2221 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg 2281 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgctgcc tcgcgcgttt 2341 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct 2401 gtaagcggat gccgtaagta aggtaaaggc ctcgcggaac ccctatttgt ttatttttct 2461 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 2521 attgaaaaag gagcttatca ccgataagga ggttttttaa tgacggtgct gcactcggtt 2581 gacttcttcc ctagcggcaa tgccagcgtt gccattgagc cgcgcctgcc tcaagccgac 2641 ttcccggagc accaccacga cttccacgag atcgttatcg tggagcacgg taccggcatc 2701 cacgttttca acggccaacc gtacacgatt acgggcggta ccgtgtgctt cgttcgtgat 2761 cacgaccgcc acttatacga gcacacggac aacttatgct taaccaacgt tttataccgt 2821 agccctgacc gcttccaatt cctggcgggc ttaaaccaac tgttaccgca ggaattagac 2881 ggccaatacc ctagccattg gcgtgtgaat cattcggtgc tgcaacaagt tcgccaatta 2941 gtggcgcaaa tggagcaaca agagggcgag aacgacctgc cgagcacggc gagccgtgaa 3001 attctgttca tgcagctgtt actgctgtta cgcaagtcga gcctgcaaga aaatttagag 3061 aattcggcga gccgcctgaa tctgctgtta gcgtggttag aagatcactt cgcggacgaa 3121 gttaactggg atgcggttgc cgaccagttc agcctgagct tacgcaccct gcaccgccaa 3181 ctgaaacaac agaccggctt aaccccgcaa cgctatttaa accgtttacg cttaatgaag 3241 gcgcgccact tactgcgcca ttcggaagcg tcggtgaccg atattgcgta ccactgtggc 3301 ttttcggata gcaatcattt cagcaccctg ttccgtcgcg aattcaattg gagccctcgc 3361 gacatccgcc aaggccgcga cggtttctta cagtgaaggc cttctagagt cgacctgcat 3421 cgacctgcag ttgacaatta atcatcggct cgtataatgt gtggaaaggc ggttcaccgc 3481 cgttttcaca caggaaacag aattctttaa gaaggagata tacatatgtc agtgatagca 3541 aagcagatga catacaaagt atatatgagc ggtactgtaa atggtcacta cttcgaagta 3601 gaaggtgatg gcaaagggaa gccgtatgag ggtgaacaaa cagtgaagct tacagttacg 3661 aagggtggcc ctttgccgtt cgcgtgggac attttgtcgc cacagtgcca gtacgggagc 3721 ataccattca ccaagtatcc agaggacata ccagactacg tgaaacagtc ctttcccgaa 3781 ggctatacct gggagcgcat aatgaacttt gaagacggtg cggtttgtac ggtatcgaac 3841 gattcatcaa tccaaggaaa ttgctttatt tatcatgtta aattttcggg gcttaacttt 3901 ccgccaaatg gccccgtgat gcagaagaaa actcagggct gggaaccaaa taccgagcgc 3961 cttttcgctc gggacggaat gctgttggga aacaatttta tggcgttgaa gctggaaggg 4021 ggtggccact atctctgtga attcaagaca acatacaaag ccaagaagcc tgtcaagatg 4081 cccggttatc actatgtgga ccggaagctc gatgtcacta atcacaacaa ggattatact 4141 tcagttgaac agtgcgaaat ttcgatcgct cgcaaacctg tcgtagcata aggggcatgc 4201 aagcttggct gttttggcgg atgagagggt accgcatgag cttcgagcta atcgcgcaaa 4261 aaaccccgct tcggcggggt tttttcgttg atcacgtacg atatcttcga accggtgcac 4321 atggtgtaca gggccctagg ataggacgtc ttaaggttta aaccaggtta attaatttaa 4381 atgcatcccg ggacgtctcg agctcgatcg gaccgcggcc gctagcacgt ataccaagtg 4441 tcctgttttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctc tgttgatacc 4501 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc 4561 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt 4621 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata 4681 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat 4741 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac 4801 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaatta 4861 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc 4921 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc 4981 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat 5041 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc 5101 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg 5161 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat 5221 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat 5281 gagtggcagg gcggggcgta atttttttaa ggcagttatt gtcttcaaat tcccgttgca 5341 tcgatggccc ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca 5401 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc 5461 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca 5521 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca 5581 gaaggccatc ctgacggatg gcctttttgc gtggccagtg ccaagcttgc atgcctgcag 5641 ctgtccggaa ctagtcgagg ccatggtggc ctcctgatgt cgtcaacacg gcgaaatagt 5701 aatcacgagg tcaggttctt accttaaatt ttcgacggaa aaccacgtaa aaaacgtcga 5761 tttttcaaga tacagcgtga attttcagga aatgcggtga gcatcacatc accacaattc 5821 agcaaattgt gaacatcatc acgttcatct ttccctggtt gccaatggcc cattttcctg 5881 tcagtaacga gaaggtcgcg aattcaggcg ctttttagac tggtcgtaat gaaattcagc 5941 aggatcacca tatggaaaaa cgcacaccac atacacgttt gagtcaggtt aaaaaacttg 6001 tcaatgccgg gcaagttcgt acaacacgta gtgccctgtt aaatgcagat gagttaggtt 6061 tggattttga tggtatgtgt aatgttatca ttggattatc agagagcgac ttttataaaa 6121 gcatgaccac ctactctgat catactatct ggcaggatgt ttacagaccc aggcttgtta 6181 caggccaggt ttatcttaaa attacggtaa ttcatgacgt actgatcgtc tcgtttaagg 6241 agaagtaagg atccgcgaat tctcatgttt gacagcttat cactgatcag tgaattaatg 6301 gcgatgacgc atcctcacga taatatccgg gtaggcgcaa tcactttcgt ctctactccg 6361 ttacaaagcg aggctgggta tttcccggcc tttctgttat ccgaaatcca ctgaaagcac 6421 agcggctggc tgaggagata aataataaac gaggggctgt atgcacaaag catcttctgt 6481 tgagttaaga acgagtatcg agatggcaca tagccttgct caaattggaa tcaggtttgt 6541 gccaatacca gtagaaacag acgaagaagt cgtgggggaa ttccacaaat tgttatccgc 6601 tcacaattcc acatgtggaa ttccacatgt ggaattccca //