LOCUS       sequence-224240-        6774 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 6774)
  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 6774)
  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..6774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(399..416)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      569..655
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      747..774
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1542..1561
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1628..1730)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1847..1865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3123..3894
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3895..4557
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     promoter        join(5057..5062,5063..5079,5080..5085)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5411..5647
                     /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     5671..5690
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5870..6748)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcaggag gaattcacca tggtacccgg ggatcctcta gagtcgacct gcaggcatgc
      361 aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
      421 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
      481 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
      541 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
      601 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
      661 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
      721 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
      781 tctacaaact cttccactca ctacacagag ccatttaaac aacatcccct ccccctttcc
      841 accgcgtcag acgcccgtag cagcccgcta cgggcttttt catgccctgc cctagcgtcc
      901 aagcctcacg gccgcgctcg gcctctctgg cggccttctg gcgctgaggt ctgcctcgtg
      961 aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga
     1021 gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt
     1081 gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag
     1141 caaaagttcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca
     1201 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     1261 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1321 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1381 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1441 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gctaaaaagg
     1501 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     1561 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     1621 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     1681 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
     1741 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
     1801 ctgcgaagtg atcttccgtc acaggtattt attcgaagac gaaagggcct cgtgatacgc
     1861 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1921 cggggaaatg tgcgcgcccg cgttcctgct ggcgctgggc ctgtttctgg cgctggactt
     1981 cccgctgttc cgtcagcagc ttttcgccca cggccttgat gatcgcggcg gccttggcct
     2041 gcatatcccg attcaacggc cccagggcgt ccagaacggg cttcaggcgc tcccgaaggt
     2101 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
     2161 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
     2221 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
     2281 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
     2341 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
     2401 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
     2461 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
     2521 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
     2581 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
     2641 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
     2701 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
     2761 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
     2821 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
     2881 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
     2941 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
     3001 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
     3061 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     3121 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     3181 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     3241 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     3301 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     3361 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     3421 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc
     3481 ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg
     3541 gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag
     3601 ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa
     3661 gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt
     3721 acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct
     3781 gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag
     3841 ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc
     3901 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
     3961 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
     4021 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
     4081 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
     4141 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
     4201 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
     4261 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
     4321 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
     4381 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
     4441 acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
     4501 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac
     4561 ttgggttgcg cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc
     4621 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     4681 gacctgaatg gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg
     4741 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     4801 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     4861 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     4921 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
     4981 aggcgtttaa gtcgaccaat aacctacggg gtctgacgct cagtggaacg aaatcgatga
     5041 gctcgcacga acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg
     5101 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     5161 ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc gggcatccaa
     5221 gtagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     5281 gcagcagggc agtcgcccta aaacaaagtt aggcagccgt tgtgctggtg ctttctgata
     5341 gttgttgtgg ggtaggcagt cagagctcga tttgcttgtc gccataatag attcacaaga
     5401 aggattcgac atgggtcaaa gtagcgatga agccaacgct cccgttgcag ggcagtttgc
     5461 gcttcccctg agtgccacct ttggcttagg ggatcgcgta cgcaagaaat ctggtgccgc
     5521 ttggcagggt caagtcgtcg gttggtattg cacaaaactc actcctgaag gctatgcggt
     5581 cgagtccgaa tcccacccag gctcagtgca aatttatcct gtggctgcac ttgaacgtgt
     5641 ggcctaagcg atatcttagg atctcccatc ggtgatgtcg gcgatatagg cgccagcaac
     5701 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg
     5761 tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     5821 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     5881 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     5941 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     6001 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     6061 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     6121 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     6181 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     6241 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     6301 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     6361 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     6421 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     6481 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     6541 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     6601 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     6661 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     6721 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agag
//
