LOCUS       sequence-214518-        7518 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 7518)
  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 7518)
  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..7518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(121..140)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(164..400)
                     /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(727..732,733..749,750..755))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(786..1634)
                     /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(1708..2544)
                     /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        2690..2808
                     /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(2915..2932)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3085..3171
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3263..3290
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4059..4078
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4145..4247)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4364..4382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5640..6409
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6410..7072
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 tctacggggt ctgacgctca gtggaacgaa atcgatgata agctgtcaaa catgagcaga
       61 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
      121 cctatatcgc cgacatcacc gatgggagat cctaagatat cgcttaggcc acacgttcaa
      181 gtgcagccac aggataaatt tgcactgagc ctgggtggga ttcggactcg accgcatagc
      241 cttcaggagt gagttttgtg caataccaac cgacgacttg accctgccaa gcggcaccag
      301 atttcttgcg tacgcgatcc cctaagccaa aggtggcact caggggaagc gcaaactgcc
      361 ctgcaacggg agcgttggct tcatcgctac tttgacccat gtcgaatcct tcttgtgaat
      421 ctattatggc gacaagcaaa tcgagctctg actgcctacc ccacaacaac tactagaaag
      481 caccagcaca acggctgcct aactttgttt tagggcgact accctgctgc gtaacatcgt
      541 tgctgttcca taacatcaaa catcgaccca cggcgtaacg cgcttagcta gcttggatgc
      601 ccgaggctag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg
      661 ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg
      721 cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcgag ctcatcgata
      781 tgcatttaat ctttctgcga attgagatga cgccactggc tgggcgtcat cccggtttcc
      841 cgggtaaaca ccaccgaaaa atagttacta tcttcaaagc cacattcggt cgaaatatca
      901 ctgattaaca ggcggctatg ctggagaaga tattgcgcat gacacactct gacctgtcgc
      961 agatattgat tgatggtcat tccagtctgc tggcgaaatt gctgacgcaa aacgcgctca
     1021 ctgcacgatg cctcatcaca aaatttatcc agcgcaaagg gacttttcag gctagccgcc
     1081 agccgggtaa tcagcttatc cagcaacgtt tcgctggatg ttggcggcaa cgaatcactg
     1141 gtgtaacgat ggcgattcag caacatcacc aactgcccga acagcaactc agccatttcg
     1201 ttagcaaacg gcacatgctg actactttca tgctcaagct gaccgataac ctgccgcgcc
     1261 tgcgccatcc ccatgctacc taagcgccag tgtggttgcc ctgcgctggc gttaaatccc
     1321 ggaatcgccc cctgccagtc aagattcagc ttcagacgct ccgggcaata aataatattc
     1381 tgcaaaacca gatcgttaac ggaagcgtag gagtgtttat cgtcagcatg aatgtaaaag
     1441 agatcgccac gggtaatgcg ataagggcga tcgttgagta catgcaggcc attaccgcgc
     1501 cagacaatca ccagctcaca aaaatcatgt gtatgttcag caaagacatc ttgcggataa
     1561 cggtcagcca cagcgactgc ctgctggtcg ctggcaaaaa aatcatcttt gagaagtttt
     1621 aactgatgcg ccaccgtggc tacctcggcc agagaacgaa gttgattatt cgcaatatgg
     1681 cgtacaaata cgttgagaag attcgcgtta ttgcagaaag ccatcccgtc cctggcgaat
     1741 atcacgcggt gaccagttaa actctcggcg aaaaagcgtc gaaaagtggt tactgtcgct
     1801 gaatccacag cgataggcga tgtcagtaac gctggcctcg ctgtggcgta gcagatgtcg
     1861 ggctttcatc agtcgcaggc ggttcaggta tcgctgaggc gtcagtcccg tttgctgctt
     1921 aagctgccga tgtagcgtac gcagtgaaag agaaaattga tccgccacgg catcccaatt
     1981 cacctcatcg gcaaaatggt cctccagcca ggccagaagc aagttgagac gtgatgcgct
     2041 gttttccagg ttctcctgca aactgctttt acgcagcaag agcagtaatt gcataaacaa
     2101 gatctcgcga ctggcggtcg agggtaaatc attttcccct tcctgctgtt ccatctgtgc
     2161 aaccagctgt cgcacctgct gcaatacgct gtggttaacg cgccagtgag acggatactg
     2221 cccatccagc tcttgtggca gcaactgatt cagcccggcg agaaactgaa atcgatccgg
     2281 cgagcgatac agcacattgg tcagacacag attatcggta tgttcataca gatgccgatc
     2341 atgatcgcgt acgaaacaga ccgtgccacc ggtgatggta tagggctgcc cattaaacac
     2401 atgaataccc gtgccatgtt cgacaatcac aatttcatga aaatcatgat gatgttcagg
     2461 aaaatccgcc tgcgggagcc ggggttctat cgccacggac gcgttaccag acggaaaaaa
     2521 atccacacta tgtaatacgg tcatactggc ctcctgatgt cgtcaacacg gcgaaatagt
     2581 aatcacgagg tcaggttctt accttaaatt ttcgacggaa aaccacgtaa aaaacgtcga
     2641 tttttcaaga tacagcgtga attttcagga aatgcggtga gcatcacatc accacaattc
     2701 agcaaattgt gaacatcatc acgttcatct ttccctggtt gccaatggcc cattttcctg
     2761 tcagtaacga gaaggtcgcg aattcaggcg ctttttagac tggtcgtaat gaaattcagc
     2821 aggatcacat atgccatggt acccggggat cctctagagt cgacctgcag gcatgcaagc
     2881 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     2941 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     3001 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     3061 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     3121 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     3181 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     3241 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     3301 caaactcttc cactcactac agcagagcca tgtagacaac atcccctccc cctttccacc
     3361 gcgtcagacg cccgtagcag cccgctacgg gctttttcat gccctgccct agcgtccaag
     3421 cctcacggcc gcgctcggcc tctctggcgg ccttctggcg ctgaggtctg cctcgtgaag
     3481 aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg
     3541 agccacggtt gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct
     3601 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
     3661 aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     3721 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     3781 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3841 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3901 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3961 cgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct aaaaaggccg
     4021 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     4081 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     4141 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     4201 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     4261 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     4321 cgaagtgatc ttccgtcaca ggtatttatt cgaagacgaa agggcctcgt gatacgccta
     4381 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4441 ggaaatgtgc gcgcccgcgt tcctgctggc gctgggcctg tttctggcgc tggacttccc
     4501 gctgttccgt cagcagcttt tcgcccacgg ccttgatgat cgcggcggcc ttggcctgca
     4561 tatcccgatt caacggcccc agggcgtcca gaacgggctt caggcgctcc cgaaggtctc
     4621 gggccgtctc ttgggcttga tcggccttct tgcgcatctc acgcgctcct gcggcggcct
     4681 gtagggcagg ctcatacccc tgccgaaccg cttttgtcag ccggtcggcc acggcttccg
     4741 gcgtctcaac gcgctttgag attcccagct tttcggccaa tccctgcggt gcataggcgc
     4801 gtggctcgac cgcttgcggg ctgatggtga cgtggcccac tggtggccgc tccagggcct
     4861 cgtagaacgc ctgaatgcgc gtgtgacgtg ccttgctgcc ctcgatgccc cgttgcagcc
     4921 ctagatcggc cacagcggcc gcaaacgtgg tctggtcgcg ggtcatctgc gctttgttgc
     4981 cgatgaactc cttggccgac agcctgccgt cctgcgtcag cggcaccacg aacgcggtca
     5041 tgtgcgggct ggtttcgtca cggtggatgc tggccgtcac gatgcgatcc gccccgtact
     5101 tgtccgccag ccacttgtgc gccttctcga agaacgccgc ctgctgttct tggctggccg
     5161 acttccacca ttccgggctg gccgtcatga cgtactcgac cgccaacaca gcgtccttgc
     5221 gccgcttctc tggcagcaac tcgcgcagtc ggcccatcgc ttcatcggtg ctgctggccg
     5281 cccagtgctc gttctctggc gtcctgctgg cgtcagcgtt gggcgtctcg cgctcgcggt
     5341 aggcgtgctt gagactggcc gccacgttgc ccattttcgc cagcttcttg catcgcatga
     5401 tcgcgtatgc cgccatgcct gcccctccct tttggtgtcc aaccggctcg acgggggcag
     5461 cgcaaggcgg tgcctccggc gggccactca atgcttgagt atactcacta gactttgctt
     5521 cgcaaagtcg tgaccgccta cggcggctgc ggcgccctac gggcttgctc tccgggcttc
     5581 gccctgcgcg gtcgctgcgc tcccttgcca gcccgtggat atgtggacga tggccgcgag
     5641 cggccaccgg ctggctcgct tcgctcggcc cgtggacaac cctgctggac aagctgatgg
     5701 acaggctgcg cctgcccacg agcttgacca cagggattgc ccaccggcta cccagccttc
     5761 gaccacatac ccaccggctc caactgcgcg gcctgcggcc ttgccccatc aattttttta
     5821 attttctctg gggaaaagcc tccggcctgc ggcctgcgcg cttcgcttgc cggttggaca
     5881 ccaagtggaa ggcgggtcaa ggctcgcgca gcgaccgcgc agcggcttgg ccttgacgcg
     5941 cctggaacga cccaagccta tgcgagtggg ggcagtcgaa ggcgaagccc gcccgcctgc
     6001 cccccgagcc tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa
     6061 ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg ccactttttg ccggaggggg
     6121 agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc accaaagaac
     6181 tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc
     6241 aacagctcat tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa
     6301 ttgactgcca cttttacgca acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga
     6361 ttgcgcatgg tgccgcaacc gtgcggcacc ctaccgcatg gagataagca tggccacgca
     6421 gtccagagaa atcggcattc aagccaagaa caagcccggt cactgggtgc aaacggaacg
     6481 caaagcgcat gaggcgtggg ccgggcttat tgcgaggaaa cccacggcgg caatgctgct
     6541 gcatcacctc gtggcgcaga tgggccacca gaacgccgtg gtggtcagcc agaagacact
     6601 ttccaagctc atcggacgtt ctttgcggac ggtccaatac gcagtcaagg acttggtggc
     6661 cgagcgctgg atctccgtcg tgaagctcaa cggccccggc accgtgtcgg cctacgtggt
     6721 caatgaccgc gtggcgtggg gccagccccg cgaccagttg cgcctgtcgg tgttcagtgc
     6781 cgccgtggtg gttgatcacg acgaccagga cgaatcgctg ttggggcatg gcgacctgcg
     6841 ccgcatcccg accctgtatc cgggcgagca gcaactaccg accggccccg gcgaggagcc
     6901 gcccagccag cccggcattc cgggcatgga accagacctg ccagccttga ccgaaacgga
     6961 ggaatgggaa cggcgcgggc agcagcgcct gccgatgccc gatgagccgt gttttctgga
     7021 cgatggcgag ccgttggagc cgccgacacg ggtcacgctg ccgcgccggt agcacttggg
     7081 ttgcgcagca acccgtaagt gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc
     7141 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     7201 gaatggaagc cggcggcacc tcgctaacgg attcaccgtt tttatcaggc tctgggaggc
     7261 agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
     7321 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     7381 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     7441 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaccaggcg
     7501 tttaagggca ccaataac
//
