LOCUS       sequence-210998-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector for Pseudomonas aeruginosa with rhaSR-PrhaBAD 
            inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/AEM.02041-16"; journalName: "Appl Environ Microbiol"; date:
            "2016-10-27- 27"; volume: "82"; issue: "22"; pages: "6715-6727"
COMMENT     Sequence Label: pJM253 (miniCTX1-rhaSR-PrhaBAD)
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(7706..7711,7712..7715,1..13,14..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
//
