LOCUS       sequence-228422-        6124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector optimized for constitutive gene expression in Burkholderia 
            genus and related bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6124)
  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 6124)
  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: pMLS7
FEATURES             Location/Qualifiers
     source          1..6124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(280..297)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      450..536
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      628..655
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1424..1443
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1510..1612)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1729..1747)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3005..3774
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3775..4437
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             complement(4889..5125)
                     /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(5452..5457,5458..5474,5475..5480))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ctggcaacca attagccggt cagtagtttc aggtcgcctg caaaggcggt aatagcggtg
       61 gtgccggaat cgccggtgct gtcgagtaag tggtcacccg accaggctgg taagtcgtta
      121 gggatgaatc gggttagttg gtggccgcgg ggctaaaagt tagctccaac tgaaaagtta
      181 aaggaagaaa cgaattcacc atggtacccg gggatcctct agagtcgacc tgcaggcatg
      241 caagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
      301 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
      361 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
      421 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
      481 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
      541 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
      601 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
      661 ttctacaaac tcttccactc actacagcag agccatgtag acaacatccc ctcccccttt
      721 ccaccgcgtc agacgcccgt agcagcccgc tacgggcttt ttcatgccct gccctagcgt
      781 ccaagcctca cggccgcgct cggcctctct ggcggccttc tggcgctgag gtctgcctcg
      841 tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt
      901 gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt
      961 ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc
     1021 agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc
     1081 cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     1141 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1201 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1261 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1321 tcaagcgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagctaaaaa
     1381 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     1441 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     1501 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     1561 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     1621 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     1681 ttctgcgaag tgatcttccg tcacaggtat ttattcgaag acgaaagggc ctcgtgatac
     1741 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1801 ttcggggaaa tgtgcgcgcc cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac
     1861 ttcccgctgt tccgtcagca gcttttcgcc cacggccttg atgatcgcgg cggccttggc
     1921 ctgcatatcc cgattcaacg gccccagggc gtccagaacg ggcttcaggc gctcccgaag
     1981 gtctcgggcc gtctcttggg cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc
     2041 ggcctgtagg gcaggctcat acccctgccg aaccgctttt gtcagccggt cggccacggc
     2101 ttccggcgtc tcaacgcgct ttgagattcc cagcttttcg gccaatccct gcggtgcata
     2161 ggcgcgtggc tcgaccgctt gcgggctgat ggtgacgtgg cccactggtg gccgctccag
     2221 ggcctcgtag aacgcctgaa tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg
     2281 cagccctaga tcggccacag cggccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt
     2341 gttgccgatg aactccttgg ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc
     2401 ggtcatgtgc gggctggttt cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc
     2461 gtacttgtcc gccagccact tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct
     2521 ggccgacttc caccattccg ggctggccgt catgacgtac tcgaccgcca acacagcgtc
     2581 cttgcgccgc ttctctggca gcaactcgcg cagtcggccc atcgcttcat cggtgctgct
     2641 ggccgcccag tgctcgttct ctggcgtcct gctggcgtca gcgttgggcg tctcgcgctc
     2701 gcggtaggcg tgcttgagac tggccgccac gttgcccatt ttcgccagct tcttgcatcg
     2761 catgatcgcg tatgccgcca tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg
     2821 ggcagcgcaa ggcggtgcct ccggcgggcc actcaatgct tgagtatact cactagactt
     2881 tgcttcgcaa agtcgtgacc gcctacggcg gctgcggcgc cctacgggct tgctctccgg
     2941 gcttcgccct gcgcggtcgc tgcgctccct tgccagcccg tggatatgtg gacgatggcc
     3001 gcgagcggcc accggctggc tcgcttcgct cggcccgtgg acaaccctgc tggacaagct
     3061 gatggacagg ctgcgcctgc ccacgagctt gaccacaggg attgcccacc ggctacccag
     3121 ccttcgacca catacccacc ggctccaact gcgcggcctg cggccttgcc ccatcaattt
     3181 ttttaatttt ctctggggaa aagcctccgg cctgcggcct gcgcgcttcg cttgccggtt
     3241 ggacaccaag tggaaggcgg gtcaaggctc gcgcagcgac cgcgcagcgg cttggccttg
     3301 acgcgcctgg aacgacccaa gcctatgcga gtgggggcag tcgaaggcga agcccgcccg
     3361 cctgcccccc gagcctcacg gcggcgagtg cgggggttcc aagggggcag cgccaccttg
     3421 ggcaaggccg aaggccgcgc agtcgatcaa caagccccgg aggggccact ttttgccgga
     3481 gggggagccg cgccgaaggc gtgggggaac cccgcagggg tgcccttctt tgggcaccaa
     3541 agaactagat atagggcgaa atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg
     3601 tacgcaacag ctcattgcgg caccccccgc aatagctcat tgcgtaggtt aaagaaaatc
     3661 tgtaattgac tgccactttt acgcaacgca taattgttgt cgcgctgccg aaaagttgca
     3721 gctgattgcg catggtgccg caaccgtgcg gcaccctacc gcatggagat aagcatggcc
     3781 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
     3841 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
     3901 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
     3961 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
     4021 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
     4081 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
     4141 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
     4201 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
     4261 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
     4321 acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
     4381 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac
     4441 ttgggttgcg cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc
     4501 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     4561 gacctgaatg gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg
     4621 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     4681 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     4741 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     4801 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
     4861 aggcgtttaa gggcaccaat aacatcgctt aggccacacg ttcaagtgca gccacaggat
     4921 aaatttgcac tgagcctggg tgggattcgg actcgaccgc atagccttca ggagtgagtt
     4981 ttgtgcaata ccaaccgacg acttgaccct gccaagcggc accagatttc ttgcgtacgc
     5041 gatcccctaa gccaaaggtg gcactcaggg gaagcgcaaa ctgccctgca acgggagcgt
     5101 tggcttcatc gctactttga cccatgtcga atccttcttg tgaatctatt atggcgacaa
     5161 gcaaatcgag ctctgactgc ctaccccaca acaactacta gaaagcacca gcacaacggc
     5221 tgcctaactt tgttttaggg cgactaccct gctgcgtaac atcgttgctg ttccataaca
     5281 tcaaacatcg acccacggcg taacgcgctt agctagcttg gatgcccgag gctagactgt
     5341 acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     5401 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga
     5461 accgaacagg cttatgtcta tacagctagc cgaattgctg gaagtcgtcg ccgccgacta
     5521 agcgctcagc gagcgcctgg ccaggtcggc aggggtacga aacatcgcaa cctgttgact
     5581 cccttagtaa tttcggaata ttatgctagg ttccggttct tggaatgcct gcgcgcgaag
     5641 tctattcgtg cgccgactac cggtccggaa gttcgatgcg caatcgtttc tgctttgccc
     5701 ggaaacagat gcgtcgattt tgttcaattt caggaataaa caatgccaac catcaatcaa
     5761 ctggttcgca aaggccgcgc gtcggaaacg acgaagagca agagcccggc tttgcaggac
     5821 tgcccccagc gtcgcggcgt gtgcacccgt gtgtacacca cgacgccgaa gaagcctaac
     5881 tcggcactgc gtaaggttgc caaggttcgt ctgacgaacg gcttcgaagt gatttcttac
     5941 atcggtggtg aaggccacaa tctgcaggaa cactcggtcg tgctgattcg cggcggtcgt
     6001 gtcaaggact tgccgggtgt gcgttaccac atggttcgtg gctcgctgga tacccagggc
     6061 gtcaaggatc gtaagcaggc tcgctcgaag tacggtgcga agcgtgcaaa agctggtaaa
     6121 taag
//
