LOCUS       sequence-36186-e        6775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector optimized for inducible gene expression in Burkholderia.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6775)
  AUTHORS   Lefebre MD, Valvano MA
  TITLE     Construction and evaluation of plasmid vectors optimized for 
            constitutive and regulated gene expression in Burkholderia cepacia 
            complex isolates.
  JOURNAL   Appl Environ Microbiol. 2002 Dec;68(12):5956-64.
  PUBMED    12450816
REFERENCE   2  (bases 1 to 6775)
  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"; journalName: "Appl 
            Environ Microbiol."; date: "2002-12"; volume: "68(12)"; pages: 
            "5956-64"
COMMENT     Sequence Label: pMLBAD
FEATURES             Location/Qualifiers
     source          1..6775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(55..60,61..77,78..83)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             410..646
                     /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"
     primer_bind     670..689
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(869..1747)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1774..2058
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1981..2000
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2172..2189)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2342..2428
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2520..2547
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3316..3335
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3402..3504)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3621..3639)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      4897..5666
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5667..6329
                     /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 atcgatgagc tcgcacgaac ccagttgaca
       61 taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc
      121 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
      181 gactgttttt ttgtacagtc tagcctcggg catccaagct agctaagcgc gttacgccgt
      241 gggtcgatgt ttgatgttat ggaacagcaa cgatgttacg cagcagggta gtcgccctaa
      301 aacaaagtta ggcagccgtt gtgctggtgc tttctagtag ttgttgtggg gtaggcagtc
      361 agagctcgat ttgcttgtcg ccataataga ttcacaagaa ggattcgaca tgggtcaaag
      421 tagcgatgaa gccaacgctc ccgttgcagg gcagtttgcg cttcccctga gtgccacctt
      481 tggcttaggg gatcgcgtac gcaagaaatc tggtgccgct tggcagggtc aagtcgtcgg
      541 ttggtattgc acaaaactca ctcctgaagg ctatgcggtc gagtccgaat cccacccagg
      601 ctcagtgcaa atttatcctg tggctgcact tgaacgtgtg gcctaagcga tatcttagga
      661 tctcccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
      721 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
      781 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
      841 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
      901 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
      961 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     1021 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     1081 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     1141 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     1201 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     1261 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     1321 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     1381 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     1441 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     1501 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     1561 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     1621 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     1681 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     1741 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
     1801 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     1861 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     1921 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     1981 atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac
     2041 tctctactgt ttctccatac ccgttttttt gggctagcag gaggaattca ccatggtacc
     2101 cggggatcct ctagagtcga cctgcaggca tgcaagcttg gctgttttgg cggatgagag
     2161 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     2221 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     2281 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2341 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2401 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     2461 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     2521 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttccac tcactacagc
     2581 agagccattt aaacaacatc ccctccccct ttccaccgcg tcagacgccc gtagcagccc
     2641 gctacgggct ttttcatgcc ctgccctagc gtccaagcct cacggccgcg ctcggcctct
     2701 ctggcggcct tctggcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
     2761 gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat gagagctttg
     2821 ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg
     2881 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc
     2941 cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     3001 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     3061 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     3121 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     3181 ctattaattt cccctcgtca aaaataaggt tatcaagcga gaaatcacca tgagtgacga
     3241 ctgaatccgg tgagaatggc aaaagctaaa aaggccgtaa tatccagctg aacggtctgg
     3301 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     3361 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     3421 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     3481 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc
     3541 ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt
     3601 atttattcga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     3661 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg cccgcgttcc
     3721 tgctggcgct gggcctgttt ctggcgctgg acttcccgct gttccgtcag cagcttttcg
     3781 cccacggcct tgatgatcgc ggcggccttg gcctgcatat cccgattcaa cggccccagg
     3841 gcgtccagaa cgggcttcag gcgctcccga aggtctcggg ccgtctcttg ggcttgatcg
     3901 gccttcttgc gcatctcacg cgctcctgcg gcggcctgta gggcaggctc atacccctgc
     3961 cgaaccgctt ttgtcagccg gtcggccacg gcttccggcg tctcaacgcg ctttgagatt
     4021 cccagctttt cggccaatcc ctgcggtgca taggcgcgtg gctcgaccgc ttgcgggctg
     4081 atggtgacgt ggcccactgg tggccgctcc agggcctcgt agaacgcctg aatgcgcgtg
     4141 tgacgtgcct tgctgccctc gatgccccgt tgcagcccta gatcggccac agcggccgca
     4201 aacgtggtct ggtcgcgggt catctgcgct ttgttgccga tgaactcctt ggccgacagc
     4261 ctgccgtcct gcgtcagcgg caccacgaac gcggtcatgt gcgggctggt ttcgtcacgg
     4321 tggatgctgg ccgtcacgat gcgatccgcc ccgtacttgt ccgccagcca cttgtgcgcc
     4381 ttctcgaaga acgccgcctg ctgttcttgg ctggccgact tccaccattc cgggctggcc
     4441 gtcatgacgt actcgaccgc caacacagcg tccttgcgcc gcttctctgg cagcaactcg
     4501 cgcagtcggc ccatcgcttc atcggtgctg ctggccgccc agtgctcgtt ctctggcgtc
     4561 ctgctggcgt cagcgttggg cgtctcgcgc tcgcggtagg cgtgcttgag actggccgcc
     4621 acgttgccca ttttcgccag cttcttgcat cgcatgatcg cgtatgccgc catgcctgcc
     4681 cctccctttt ggtgtccaac cggctcgacg ggggcagcgc aaggcggtgc ctccggcggg
     4741 ccactcaatg cttgagtata ctcactagac tttgcttcgc aaagtcgtga ccgcctacgg
     4801 cggctgcggc gccctacggg cttgctctcc gggcttcgcc ctgcgcggtc gctgcgctcc
     4861 cttgccagcc cgtggatatg tggacgatgg ccgcgagcgg ccaccggctg gctcgcttcg
     4921 ctcggcccgt ggacaaccct gctggacaag ctgatggaca ggctgcgcct gcccacgagc
     4981 ttgaccacag ggattgccca ccggctaccc agccttcgac cacataccca ccggctccaa
     5041 ctgcgcggcc tgcggccttg ccccatcaat ttttttaatt ttctctgggg aaaagcctcc
     5101 ggcctgcggc ctgcgcgctt cgcttgccgg ttggacacca agtggaaggc gggtcaaggc
     5161 tcgcgcagcg accgcgcagc ggcttggcct tgacgcgcct ggaacgaccc aagcctatgc
     5221 gagtgggggc agtcgaaggc gaagcccgcc cgcctgcccc ccgagcctca cggcggcgag
     5281 tgcgggggtt ccaagggggc agcgccacct tgggcaaggc cgaaggccgc gcagtcgatc
     5341 aacaagcccc ggaggggcca ctttttgccg gagggggagc cgcgccgaag gcgtggggga
     5401 accccgcagg ggtgcccttc tttgggcacc aaagaactag atatagggcg aaatgcgaaa
     5461 gacttaaaaa tcaacaactt aaaaaagggg ggtacgcaac agctcattgc ggcacccccc
     5521 gcaatagctc attgcgtagg ttaaagaaaa tctgtaattg actgccactt ttacgcaacg
     5581 cataattgtt gtcgcgctgc cgaaaagttg cagctgattg cgcatggtgc cgcaaccgtg
     5641 cggcacccta ccgcatggag ataagcatgg ccacgcagtc cagagaaatc ggcattcaag
     5701 ccaagaacaa gcccggtcac tgggtgcaaa cggaacgcaa agcgcatgag gcgtgggccg
     5761 ggcttattgc gaggaaaccc acggcggcaa tgctgctgca tcacctcgtg gcgcagatgg
     5821 gccaccagaa cgccgtggtg gtcagccaga agacactttc caagctcatc ggacgttctt
     5881 tgcggacggt ccaatacgca gtcaaggact tggtggccga gcgctggatc tccgtcgtga
     5941 agctcaacgg ccccggcacc gtgtcggcct acgtggtcaa tgaccgcgtg gcgtggggcc
     6001 agccccgcga ccagttgcgc ctgtcggtgt tcagtgccgc cgtggtggtt gatcacgacg
     6061 accaggacga atcgctgttg gggcatggcg acctgcgccg catcccgacc ctgtatccgg
     6121 gcgagcagca actaccgacc ggccccggcg aggagccgcc cagccagccc ggcattccgg
     6181 gcatggaacc agacctgcca gccttgaccg aaacggagga atgggaacgg cgcgggcagc
     6241 agcgcctgcc gatgcccgat gagccgtgtt ttctggacga tggcgagccg ttggagccgc
     6301 cgacacgggt cacgctgccg cgccggtagc acttgggttg cgcagcaacc cgtaagtgcg
     6361 ctgttccaga ctatcggctg tagccgcctc gccgccctat accttgtctg cctccccgcg
     6421 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg
     6481 ctaacggatt caccgttttt atcaggctct gggaggcaga ataaatgatc atatcgtcaa
     6541 ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag aagcacacgg
     6601 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     6661 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
     6721 ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagtcgacca ataac
//
