LOCUS       sequence-211000-        8252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector for Pseudomonas aeruginosa with rhaSR-PrhaBAD 
            inducible promoter and StRBS-aacC1 insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8252)
  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 8252)
  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: pJM260 (miniCTX1-rhaSR-PrhaBAD-stRBS-aacC1)
FEATURES             Location/Qualifiers
     source          1..8252
                     /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)"
     CDS             3025..3558
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(3567..3583)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3568..3584)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3608..3627)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3609..3627)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(3801..3848)
                     /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            4055..4163
                     /note="incP origin of transfer"
     rep_origin      complement(5595..6183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5675..5694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7005..8195)
                     /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     7824..7843
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(8243..8248,8249..8252,1..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 gctctgcagt aaggaggagc aacgatgtta cgcagcagca acgatgttac gcagcagggc
     3061 agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc
     3121 ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga
     3181 gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt
     3241 agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg
     3301 gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca
     3361 gtctccggcg agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat
     3421 gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc
     3481 gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac
     3541 ccaagtaccg ccacctaaaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc
     3601 caattcgccc tatagtgagt cgtattacgc gcgccggtgg atgacctttt gaatgacctt
     3661 taatagatta tattactaat taattgggga ccctagaggt cccctttttt attttaaaaa
     3721 ttttttcaca aaacggttta caagcataaa gctaattcca ttgagtaagt ttttaagcac
     3781 atcagcttca aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata
     3841 ggtacttcaa gatccccaat tcgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4021 gggataacgc aggaaagaac atgctagagc ttatcggcca gcctcgcaga gcaggattcc
     4081 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     4141 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     4201 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     4261 ctataatgac cccgaagcag ggttatgcag cggaaagtat accttaacct ttttcgttgc
     4321 ttcaggaaag tttcagcgcc tgattcagca gacctacgat gtcggggccg tccttgctga
     4381 tccacttggc gtaatgcttg aagatcatcg ccgtcgaggt gtgccccatc tggtccgcga
     4441 tccattcggg agtggcgatg ccgctgctga gcatctggct ggcgaaggtg tggcggcact
     4501 gattgggacc acgcgagcgg acgcccgcat tcctgaggtg gccatgccac cagccgttgc
     4561 gcagcgtatc cgacgtggag tacgccgccc cgctagcact gttgtggaac acgaagcgta
     4621 cccgttgagg ctttctggtc cggttgtcgc ggtctatcac ctcgatctgt acgggcggaa
     4681 ggcgtctggt tagcttggcc tgctgctgga gtgcgcgcag cgccggggcc agtagctgga
     4741 ccttcctcgt cgatcggcga gtcttggtga ccttgtactg gctgcgtaca cgagcgcggc
     4801 ggaaaaccac ggtccccgtg tccaaatcga catcttccca ggccagagct atggcctcgc
     4861 tgacgcgggg gccgctccag atcatgaact ccgccaggtt cagctcaccg atgcgcgcgg
     4921 tttcagtacc gaggatgagg tcgatttcct cacgagtgaa tgggtctggg tcatccgcat
     4981 ccggaagagt gatcacgatg ccgtctgttg ggtcgtgcgc gctccggttc cgagttcgat
     5041 acagccggaa gatctgccgc aggttgctga cgatctctcg caccgtcttg ttgtgcagct
     5101 tgggcatgag cgtgttctgc acccagtctt gaatatccag gtggtcgatg ctgtcggcct
     5161 ggctgtcacc ccagcgaggt cggatgtggg tttccacgcg gctggtgtag ccgcggaagc
     5221 cgctggcggc aatctggttg cgtttgatgt cgagccagag gtcgatgtaa tggcccaagg
     5281 tgttgctttt cacccgcggt gagtccggga agtgtcggct gtagctgaag actccttgct
     5341 tgatctcgta gttgatgatg ccggccagct gttcggcatt ggctatattc tcgggcgttg
     5401 cgtcccctgg tatggactcc ctacacagct cgccctggta gcggaagtag atcctgatcc
     5461 gttttccgcg tacctcgact ccgtctgcca tctggttccc ctgatccaca ccaagcgttc
     5521 aggctaccaa gctggatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5581 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5641 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5701 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5761 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5821 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5881 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5941 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6001 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6061 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6121 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6181 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6241 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6301 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6361 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6421 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6481 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6541 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6601 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6661 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6721 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6781 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6841 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6901 ggtgagtcaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     6961 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     7021 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     7081 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     7141 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     7201 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     7261 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     7321 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     7381 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     7441 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     7501 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     7561 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     7621 atgggtcggc gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     7681 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgta aggagatggc gcccaacagt
     7741 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     7801 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     7861 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga gaatccacag gacgggtgtg
     7921 gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg
     7981 cggccaaagc ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca
     8041 tatagcgcta gcagcacgcc atagtgactg gcgatgctgt cggaatggac aatatcccgc
     8101 aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg
     8161 ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa
     8221 tttaactgtg ataaactacc gcattaaagc ta
//
