LOCUS Exported 7715 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Integration vector for Pseudomonas aeruginosa with rhaSR-PrhaBAD inducible promoter. ACCESSION . VERSION . KEYWORDS pJM253 (miniCTX1-rhaSR-PrhaBAD) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7715) AUTHORS Meisner J, Goldberg JB TITLE The Escherichia coli rhaSR-PrhaBAD Inducible Promoter System Allows Tightly Controlled Gene Expression over a Wide Range in Pseudomonas aeruginosa. JOURNAL Appl Environ Microbiol. 2016 Oct 27;82(22):6715-6727. doi: 10.1128/AEM.02041-16. Print 2016 Nov 15. PUBMED 27613678 REFERENCE 2 (bases 1 to 7715) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7715 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(238..285) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." primer_bind 822..842 /label=T3 /note="T3 promoter, forward primer" promoter 824..842 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS complement(962..1810) /codon_start=1 /gene="E. coli rhaR" /product="transcriptional activator of rhaR and rhaS" /label=rhaR /note="needed for rhamnose-dependent activation of the rhaB promoter" /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK D" CDS complement(1884..2720) /codon_start=1 /gene="E. coli rhaS" /product="positive regulator of the rhaB promoter " /label=rhaS /note="activated by L-rhamnose" /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ" promoter 2866..2984 /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)" primer_bind complement(3030..3046) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3031..3047) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(3071..3090) /label=T7 /note="T7 promoter, forward primer" promoter complement(3072..3090) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind complement(3264..3311) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." oriT 3518..3626 /note="incP origin of transfer" rep_origin complement(5058..5646) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5138..5157) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6468..7658) /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" primer_bind 7287..7306 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter complement(7706..19) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" ORIGIN 1 atcgatgata agctgtcaaa catgagaatt aattcatgtt ctttcctgcg ttatcccctg 61 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 181 ctctccccgc gcgttggccg attcattaat gcagcgaatt agcttcaaaa gcgctctgaa 241 gttcctatac tttctagaga ataggaactt cggaatagga acttcaagat cccctgattc 301 cctttgtcaa cagcaatgga tttcaatggg tcaacatgaa atccggctgt tgccctggga 361 cggcaggttt gacggcaggc ttccaggctc tctcgcacac gctgggaaca ctctgggaac 421 aaacgcgagt gaaaatggac agtagggccc aggaataatg cggcctccag cgaggttggg 481 gcgtttcata tgagttcgaa tctcaccgcc tccgccatat aacctgctgt tttcattgat 541 gtttccattc gacccactct caggagtgaa cgatttggga acaaactggg aacgaggcgc 601 agaaaaaagg ggccccgcag ggcccctttt tcgttgcttc aggaaagttt cagcgcctga 661 ttcagcagac ctacgatgtc ggggccgtcc ttgctgaatt agctttatgc ttgtaaaccg 721 ttttgtgaaa aaatttttaa aataaaaaag gggacctcta gggtccccaa ttaattagta 781 atataatcta ttaaaggtca ttcaaaaggt catccacccg cgcaattaac cctcactaaa 841 gggaacaaaa gctggagctc aaagagtgga acaatgcagg acgccgtccg gtaacggtgt 901 tgagcctcgg ttgtgtggtg attattgtcg ccgctaacat cgtcggcatc ggcatggcga 961 attaatcttt ctgcgaattg agatgacgcc actggctggg cgtcatcccg gtttcccggg 1021 taaacaccac cgaaaaatag ttactatctt caaagccaca ttcggtcgaa atatcactga 1081 ttaacaggcg gctatgctgg agaagatatt gcgcatgaca cactctgacc tgtcgcagat 1141 attgattgat ggtcattcca gtctgctggc gaaattgctg acgcaaaacg cgctcactgc 1201 acgatgcctc atcacaaaat ttatccagcg caaagggact tttcaggcta gccgccagcc 1261 gggtaatcag cttatccagc aacgtttcgc tggatgttgg cggcaacgaa tcactggtgt 1321 aacgatggcg attcagcaac atcaccaact gcccgaacag caactcagcc atttcgttag 1381 caaacggcac atgctgacta ctttcatgct caagctgacc gataacctgc cgcgcctgcg 1441 ccatccccat gctacctaag cgccagtgtg gttgccctgc gctggcgtta aatcccggaa 1501 tcgccccctg ccagtcaaga ttcagcttca gacgctccgg gcaataaata atattctgca 1561 aaaccagatc gttaacggaa gcgtaggagt gtttatcgtc agcatgaatg taaaagagat 1621 cgccacgggt aatgcgataa gggcgatcgt tgagtacatg caggccatta ccgcgccaga 1681 caatcaccag ctcacaaaaa tcatgtgtat gttcagcaaa gacatcttgc ggataacggt 1741 cagccacagc gactgcctgc tggtcgctgg caaaaaaatc atctttgaga agttttaact 1801 gatgcgccac cgtggctacc tcggccagag aacgaagttg attattcgca atatggcgta 1861 caaatacgtt gagaagattc gcgttattgc agaaagccat cccgtccctg gcgaatatca 1921 cgcggtgacc agttaaactc tcggcgaaaa agcgtcgaaa agtggttact gtcgctgaat 1981 ccacagcgat aggcgatgtc agtaacgctg gcctcgctgt ggcgtagcag atgtcgggct 2041 ttcatcagtc gcaggcggtt caggtatcgc tgaggcgtca gtcccgtttg ctgcttaagc 2101 tgccgatgta gcgtacgcag tgaaagagaa aattgatccg ccacggcatc ccaattcacc 2161 tcatcggcaa aatggtcctc cagccaggcc agaagcaagt tgagacgtga tgcgctgttt 2221 tccaggttct cctgcaaact gcttttacgc agcaagagca gtaattgcat aaacaagatc 2281 tcgcgactgg cggtcgaggg taaatcattt tccccttcct gctgttccat ctgtgcaacc 2341 agctgtcgca cctgctgcaa tacgctgtgg ttaacgcgcc agtgagacgg atactgccca 2401 tccagctctt gtggcagcaa ctgattcagc ccggcgagaa actgaaatcg atccggcgag 2461 cgatacagca cattggtcag acacagatta tcggtatgtt catacagatg ccgatcatga 2521 tcgcgtacga aacagaccgt gccaccggtg atggtatagg gctgcccatt aaacacatga 2581 atacccgtgc catgttcgac aatcacaatt tcatgaaaat catgatgatg ttcaggaaaa 2641 tccgcctgcg ggagccgggg ttctatcgcc acggacgcgt taccagacgg aaaaaaatcc 2701 acactatgta atacggtcat actggcctcc tgatgtcgtc aacacggcga aatagtaatc 2761 acgaggtcag gttcttacct taaattttcg acggaaaacc acgtaaaaaa cgtcgatttt 2821 tcaagataca gcgtgaattt tcaggaaatg cggtgagcat cacatcacca caattcagca 2881 aattgtgaac atcatcacgt tcatctttcc ctggttgcca atggcccatt ttcctgtcag 2941 taacgagaag gtcgcgaatt caggcgcttt ttagactggt cgtaatgaaa ttcaactagt 3001 gctctgcagg aattcctcga gaagcttatc gataccgtcg acctcgaggg ggggcccggt 3061 acccaattcg ccctatagtg agtcgtatta cgcgcgccgg tggatgacct tttgaatgac 3121 ctttaataga ttatattact aattaattgg ggaccctaga ggtccccttt tttattttaa 3181 aaattttttc acaaaacggt ttacaagcat aaagctaatt ccattgagta agtttttaag 3241 cacatcagct tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga 3301 ataggtactt caagatcccc aattcgctgc attaatgaat cggccaacgc gcggggagag 3361 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 3421 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 3481 caggggataa cgcaggaaag aacatgctag agcttatcgg ccagcctcgc agagcaggat 3541 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg 3601 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta 3661 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa 3721 tcgctataat gaccccgaag cagggttatg cagcggaaag tataccttaa cctttttcgt 3781 tgcttcagga aagtttcagc gcctgattca gcagacctac gatgtcgggg ccgtccttgc 3841 tgatccactt ggcgtaatgc ttgaagatca tcgccgtcga ggtgtgcccc atctggtccg 3901 cgatccattc gggagtggcg atgccgctgc tgagcatctg gctggcgaag gtgtggcggc 3961 actgattggg accacgcgag cggacgcccg cattcctgag gtggccatgc caccagccgt 4021 tgcgcagcgt atccgacgtg gagtacgccg ccccgctagc actgttgtgg aacacgaagc 4081 gtacccgttg aggctttctg gtccggttgt cgcggtctat cacctcgatc tgtacgggcg 4141 gaaggcgtct ggttagcttg gcctgctgct ggagtgcgcg cagcgccggg gccagtagct 4201 ggaccttcct cgtcgatcgg cgagtcttgg tgaccttgta ctggctgcgt acacgagcgc 4261 ggcggaaaac cacggtcccc gtgtccaaat cgacatcttc ccaggccaga gctatggcct 4321 cgctgacgcg ggggccgctc cagatcatga actccgccag gttcagctca ccgatgcgcg 4381 cggtttcagt accgaggatg aggtcgattt cctcacgagt gaatgggtct gggtcatccg 4441 catccggaag agtgatcacg atgccgtctg ttgggtcgtg cgcgctccgg ttccgagttc 4501 gatacagccg gaagatctgc cgcaggttgc tgacgatctc tcgcaccgtc ttgttgtgca 4561 gcttgggcat gagcgtgttc tgcacccagt cttgaatatc caggtggtcg atgctgtcgg 4621 cctggctgtc accccagcga ggtcggatgt gggtttccac gcggctggtg tagccgcgga 4681 agccgctggc ggcaatctgg ttgcgtttga tgtcgagcca gaggtcgatg taatggccca 4741 aggtgttgct tttcacccgc ggtgagtccg ggaagtgtcg gctgtagctg aagactcctt 4801 gcttgatctc gtagttgatg atgccggcca gctgttcggc attggctata ttctcgggcg 4861 ttgcgtcccc tggtatggac tccctacaca gctcgccctg gtagcggaag tagatcctga 4921 tccgttttcc gcgtacctcg actccgtctg ccatctggtt cccctgatcc acaccaagcg 4981 ttcaggctac caagctggat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 5041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 5101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 5161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 5221 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 5281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 5341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 5401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 5461 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 5521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 5581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 5641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 5701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 5761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 5821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 5881 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 5941 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 6001 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 6061 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 6121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 6181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 6241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 6301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 6361 actggtgagt caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc 6421 aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg 6481 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct 6541 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc 6601 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc 6661 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg 6721 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc 6781 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa 6841 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc 6901 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc 6961 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg 7021 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag 7081 ggcatgggtc ggcgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg 7141 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gtaaggagat ggcgcccaac 7201 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg 7261 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca 7321 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agagaatcca caggacgggt 7381 gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg 7441 cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac 7501 gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacaatatcc 7561 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg 7621 gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag 7681 caatttaact gtgataaact accgcattaa agcta //