LOCUS       sequence-220100-        7517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An expression vector containing a rhamnose-inducible promoter to 
            regulate gene expression in Burkholderia cenocepacia.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7517)
  AUTHORS   Cardona ST, Valvano MA
  TITLE     An expression vector containing a rhamnose-inducible promoter 
            provides tightly regulated gene expression in Burkholderia 
            cenocepacia.
  JOURNAL   Plasmid. 2005 Nov;54(3):219-28. doi: 10.1016/j.plasmid.2005.03.004. 
            Epub 2005 May 31.
  PUBMED    15925406
REFERENCE   2  (bases 1 to 7517)
  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: 
            "10.1016/j.plasmid.2005.03.004"; journalName: "Plasmid"; date: 
            "2005-11"; volume: "54"; issue: "3"; pages: "219-28"
COMMENT     Sequence Label: pSCrhaB2
FEATURES             Location/Qualifiers
     source          1..7517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)"
     primer_bind     complement(226..243)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      396..482
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      574..601
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1067..1729)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      1730..2501
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     primer_bind     3759..3777
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3894..3996
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4063..4082)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(4952..4971)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4995..5231)
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     promoter        complement(join(5557..5562,5563..5579,5580..5585))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(5614..6462)
                     /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(6536..7372)
                     /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"
ORIGIN
        1 caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc
       61 ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga ctggtcgtaa
      121 tgaaattcag caggatcaca tatgccatgg tacccgggga tcctctagag tcgacctgca
      181 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
      241 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
      301 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
      361 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
      421 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      481 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
      541 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
      601 ttgcgtttct acaaactctt gttattggtc gacttaaacg cctggtgcta cgcctgaata
      661 agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt
      721 cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt ttattgacta
      781 ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct caggctctcc
      841 ccgtggaggt aataattgac gatatgatca tttattctgc ctcccagagc ctgataaaaa
      901 cggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
      961 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgaggc ggctacagcc
     1021 gatagtctgg aacagcgcac ttacgggttg ctgcgcaacc caagtgctac cggcgcggca
     1081 gcgtgacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac ggctcatcgg
     1141 gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc aaggctggca
     1201 ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg gggccggtcg
     1261 gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca tgccccaaca
     1321 gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc gacaggcgca
     1381 actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc gacacggtgc
     1441 cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc ttgactgcgt
     1501 attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgtcttctgg ctgaccacca
     1561 cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc gccgtgggtt
     1621 tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc acccagtgac
     1681 cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg cttatctcca
     1741 tgcggtaggg gtgccgcacg gttgcggcac catgcgcaat cagctgcaac ttttcggcag
     1801 cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt ctttaaccta
     1861 cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc ttttttaagt
     1921 tgttgatttt taagtctttc gcatttcgcc ctatatctag ttctttggtg cccaaagaag
     1981 ggcacccctg cggggttccc ccacgccttc ggcgcggctc cccctccggc aaaaagtggc
     2041 ccctccgggg cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt ggcgctgccc
     2101 ccttggaacc cccgcactcg ccgccgtgag gctcgggggg caggcgggcg ggcttcgccc
     2161 ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg ccaagccgct
     2221 gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg gcaagcgaag
     2281 cgcgcaggcc gcaggccgga ggcttttccc cagagaaaat taaaaaaatt gatggggcaa
     2341 ggccgcaggc cgcgcagttg gagccggtgg gtatgtggtc gaaggctggg tagccggtgg
     2401 gcaatccctg tggtcaagct cgtgggcagg cgcagcctgt ccatcagctt gtccagcagg
     2461 gttgtccacg ggccgagcga agcgagccag ccggtggccg ctcgcggcca tcgtccacat
     2521 atccacgggc tggcaaggga gcgcagcgac cgcgcagggc gaagcccgga gagcaagccc
     2581 gtagggcgcc gcagccgccg taggcggtca cgactttgcg aagcaaagtc tagtgagtat
     2641 actcaagcat tgagtggccc gccggaggca ccgccttgcg ctgcccccgt cgagccggtt
     2701 ggacaccaaa agggaggggc aggcatggcg gcatacgcga tcatgcgatg caagaagctg
     2761 gcgaaaatgg gcaacgtggc ggccagtctc aagcacgcct accgcgagcg cgagacgccc
     2821 aacgctgacg ccagcaggac gccagagaac gagcactggg cggccagcag caccgatgaa
     2881 gcgatgggcc gactgcgcga gttgctgcca gagaagcggc gcaaggacgc tgtgttggcg
     2941 gtcgagtacg tcatgacggc cagcccggaa tggtggaagt cggccagcca agaacagcag
     3001 gcggcgttct tcgagaaggc gcacaagtgg ctggcggaca agtacggggc ggatcgcatc
     3061 gtgacggcca gcatccaccg tgacgaaacc agcccgcaca tgaccgcgtt cgtggtgccg
     3121 ctgacgcagg acggcaggct gtcggccaag gagttcatcg gcaacaaagc gcagatgacc
     3181 cgcgaccaga ccacgtttgc ggccgctgtg gccgatctag ggctgcaacg gggcatcgag
     3241 ggcagcaagg cacgtcacac gcgcattcag gcgttctacg aggccctgga gcggccacca
     3301 gtgggccacg tcaccatcag cccgcaagcg gtcgagccac gcgcctatgc accgcaggga
     3361 ttggccgaaa agctgggaat ctcaaagcgc gttgagacgc cggaagccgt ggccgaccgg
     3421 ctgacaaaag cggttcggca ggggtatgag cctgccctac aggccgccgc aggagcgcgt
     3481 gagatgcgca agaaggccga tcaagcccaa gagacggccc gagaccttcg ggagcgcctg
     3541 aagcccgttc tggacgccct ggggccgttg aatcgggata tgcaggccaa ggccgccgcg
     3601 atcatcaagg ccgtgggcga aaagctgctg acggaacagc gggaagtcca gcgccagaaa
     3661 caggcccagc gccagcagga acgcgggcgc gcacatttcc ccgaaaagtg ccacctgacg
     3721 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3781 ttcgtcttcg aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt
     3841 gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg
     3901 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     3961 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     4021 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     4081 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt agcttttgcc
     4141 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     4201 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     4261 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     4321 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     4381 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     4441 gacttgacgg gacggcggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     4501 atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc
     4561 aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt ctggctggat
     4621 gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg cagacctcag
     4681 cgccagaagg ccgccagaga ggccgagcgc ggccgtgagg cttggacgct agggcagggc
     4741 atgaaaaagc ccgtagcggg ctgctacggg cgtctgacgc ggtggaaagg gggaggggat
     4801 gttgtttaaa tggctctgct gtagtgagtg gtctacgggg tctgacgctc agtggaacga
     4861 aatcgatgat aagctgtcaa acatgagcag atcctctacg ccggacgcat cgtggccggc
     4921 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatgggaga
     4981 tcctaagata tcgcttaggc cacacgttca agtgcagcca caggataaat ttgcactgag
     5041 cctgggtggg attcggactc gaccgcatag ccttcaggag tgagttttgt gcaataccaa
     5101 ccgacgactt gaccctgcca agcggcacca gatttcttgc gtacgcgatc ccctaagcca
     5161 aaggtggcac tcaggggaag cgcaaactgc cctgcaacgg gagcgttggc ttcatcgcta
     5221 ctttgaccca tgtcgaatcc ttcttgtgaa tctattatgg cgacaagcaa atcgagctct
     5281 gactgcctac cccacaacaa ctatcagaaa gcaccagcac aacggctgcc taactttgtt
     5341 ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     5401 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     5461 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     5521 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     5581 gtcaactggg ttcgtgcgag ctcatcgatg catttaatct ttctgcgaat tgagatgacg
     5641 ccactggctg ggcgtcatcc cggtttcccg ggtaaacacc accgaaaaat agttactatc
     5701 ttcaaagcca cattcggtcg aaatatcact gattaacagg cggctatgct ggagaagata
     5761 ttgcgcatga cacactctga cctgtcgcag atattgattg atggtcattc cagtctgctg
     5821 gcgaaattgc tgacgcaaaa cgcgctcact gcacgatgcc tcatcacaaa atttatccag
     5881 cgcaaaggga cttttcaggc tagccgccag ccgggtaatc agcttatcca gcaacgtttc
     5941 gctggatgtt ggcggcaacg aatcactggt gtaacgatgg cgattcagca acatcaccaa
     6001 ctgcccgaac agcaactcag ccatttcgtt agcaaacggc acatgctgac tactttcatg
     6061 ctcaagctga ccgataacct gccgcgcctg cgccatcccc atgctaccta agcgccagtg
     6121 tggttgccct gcgctggcgt taaatcccgg aatcgccccc tgccagtcaa gattcagctt
     6181 cagacgctcc gggcaataaa taatattctg caaaaccaga tcgttaacgg aagcgtagga
     6241 gtgtttatcg tcagcatgaa tgtaaaagag atcgccacgg gtaatgcgat aagggcgatc
     6301 gttgagtaca tgcaggccat taccgcgcca gacaatcacc agctcacaaa aatcatgtgt
     6361 atgttcagca aagacatctt gcggataacg gtcagccaca gcgactgcct gctggtcgct
     6421 ggcaaaaaaa tcatctttga gaagttttaa ctgatgcgcc accgtggcta cctcggccag
     6481 agaacgaagt tgattattcg caatatggcg tacaaatacg ttgagaagat tcgcgttatt
     6541 gcagaaagcc atcccgtccc tggcgaatat cacgcggtga ccagttaaac tctcggcgaa
     6601 aaagcgtcga aaagtggtta ctgtcgctga atccacagcg ataggcgatg tcagtaacgc
     6661 tggcctcgct gtggcgtagc agatgtcggg ctttcatcag tcgcaggcgg ttcaggtatc
     6721 gctgaggcgt cagtcccgtt tgctgcttaa gctgccgatg tagcgtacgc agtgaaagag
     6781 aaaattgatc cgccacggca tcccaattca cctcatcggc aaaatggtcc tccagccagg
     6841 ccagaagcaa gttgagacgt gatgcgctgt tttccaggtt ctcctgcaaa ctgcttttac
     6901 gcagcaagag cagtaattgc ataaacaaga tctcgcgact ggcggtcgag ggtaaatcat
     6961 tttccccttc ctgctgttcc atctgtgcaa ccagctgtcg cacctgctgc aatacgctgt
     7021 ggttaacgcg ccagtgagac ggatactgcc catccagctc ttgtggcagc aactgattca
     7081 gcccggcgag aaactgaaat cgatccggcg agcgatacag cacattggtc agacacagat
     7141 tatcggtatg ttcatacaga tgccgatcat gatcgcgtac gaaacagacc gtgccaccgg
     7201 tgatggtata gggctgccca ttaaacacat gaatacccgt gccatgttcg acaatcacaa
     7261 tttcatgaaa atcatgatga tgttcaggaa aatccgcctg cgggagccgg ggttctatcg
     7321 ccacggacgc gttaccagac ggaaaaaaat ccacactatg taatacggtc atactggcct
     7381 cctgatgtcg tcaacacggc gaaatagtaa tcacgaggtc aggttcttac cttaaatttt
     7441 cgacggaaaa ccacgtaaaa aacgtcgatt tttcaagata cagcgtgaat tttcaggaaa
     7501 tgcggtgagc atcacat
//
