LOCUS       sequence-220094-        6180 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 6180)
  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 6180)
  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..6180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(282..299)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      452..538
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      630..657
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1425..1444
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1511..1613)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1730..1748)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3006..3777
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3778..4440
                     /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(4892..5128)
                     /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(5454..5459,5460..5476,5477..5482))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 agctggcaac caattagccg gtcagtagtt tcaggtcgcc tgcaaaggcg gtaatagcgg
       61 tggtgccgga atcgccggtg ctgtcgagta agtggtcacc cgaccaggct ggtaagtcgt
      121 tagggatgaa tcgggttagt tggtggccgc ggggctaaaa gttagctcca actgaaaagt
      181 taaaggaaga aacgaattca ccatggtacc cggggatcct ctagagtcga cctgcaggca
      241 tgcaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
      301 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
      361 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
      421 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      481 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
      541 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
      601 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
      661 gtttctacaa actctccact cactacagca gagccattta aacaacatcc cctccccctt
      721 tccaccgcgt cagacgcccg tagcagcccg ctacgggctt tttcatgccc tgccctagcg
      781 tccaagcctc acggccgcgc tcggcctctc tggcggcctt ctggcgctga ggtctgcctc
      841 gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc agccagaaag
      901 tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg attttgaact
      961 tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact
     1021 cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg taatgctctg
     1081 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     1141 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1201 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1261 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1321 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagctaaaa
     1381 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     1441 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     1501 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     1561 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc
     1621 attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt
     1681 attctgcgaa gtgatcttcc gtcacaggta tttattcgaa gacgaaaggg cctcgtgata
     1741 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     1801 tttcggggaa atgtgcgcgc ccgcgttcct gctggcgctg ggcctgtttc tggcgctgga
     1861 cttcccgctg ttccgtcagc agcttttcgc ccacggcctt gatgatcgcg gcggccttgg
     1921 cctgcatatc ccgattcaac ggccccaggg cgtccagaac gggcttcagg cgctcccgaa
     1981 ggtctcgggc cgtctcttgg gcttgatcgg ccttcttgcg catctcacgc gctcctgcgg
     2041 cggcctgtag ggcaggctca tacccctgcc gaaccgcttt tgtcagccgg tcggccacgg
     2101 cttccggcgt ctcaacgcgc tttgagattc ccagcttttc ggccaatccc tgcggtgcat
     2161 aggcgcgtgg ctcgaccgct tgcgggctga tggtgacgtg gcccactggt ggccgctcca
     2221 gggcctcgta gaacgcctga atgcgcgtgt gacgtgcctt gctgccctcg atgccccgtt
     2281 gcagccctag atcggccaca gcggccgcaa acgtggtctg gtcgcgggtc atctgcgctt
     2341 tgttgccgat gaactccttg gccgacagcc tgccgtcctg cgtcagcggc accacgaacg
     2401 cggtcatgtg cgggctggtt tcgtcacggt ggatgctggc cgtcacgatg cgatccgccc
     2461 cgtacttgtc cgccagccac ttgtgcgcct tctcgaagaa cgccgcctgc tgttcttggc
     2521 tggccgactt ccaccattcc gggctggccg tcatgacgta ctcgaccgcc aacacagcgt
     2581 ccttgcgccg cttctctggc agcaactcgc gcagtcggcc catcgcttca tcggtgctgc
     2641 tggccgccca gtgctcgttc tctggcgtcc tgctggcgtc agcgttgggc gtctcgcgct
     2701 cgcggtaggc gtgcttgaga ctggccgcca cgttgcccat tttcgccagc ttcttgcatc
     2761 gcatgatcgc gtatgccgcc atgcctgccc ctcccttttg gtgtccaacc ggctcgacgg
     2821 gggcagcgca aggcggtgcc tccggcgggc cactcaatgc ttgagtatac tcactagact
     2881 ttgcttcgca aagtcgtgac cgcctacggc ggctgcggcg ccctacgggc ttgctctccg
     2941 ggcttcgccc tgcgcggtcg ctgcgctccc ttgccagccc gtggatatgt ggacgatggc
     3001 cgcgagcggc caccggctgg ctcgcttcgc tcggcccgtg gacaaccctg ctggacaagc
     3061 tgatggacag gctgcgcctg cccacgagct tgaccacagg gattgcccac cggctaccca
     3121 gccttcgacc acatacccac cggctccaac tgcgcggcct gcggccttgc cccatcaatt
     3181 tttttaattt tctctgggga aaagcctccg gcctgcggcc tgcgcgcttc gcttgccggt
     3241 tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga ccgcgcagcg gcttggcctt
     3301 gacgcgcctg gaacgaccca agcctatgcg agtgggggca gtcgaagggc gaagcccgcc
     3361 cgcctgcccc ccgagcctca cggcggcgag tgcgggggtt ccaagggggc agcgccacct
     3421 tgggcaaggc cgaaggccgc gcagtcgatc aacaagcccc ggaggggcca ctttttgccg
     3481 gagggggagc cgcgccgaag gcgtggggga accccgcagg ggtgcccttc tttgggcacc
     3541 aaagaactag atatagggcg aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg
     3601 ggtacgcaac agctcattgc ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa
     3661 tctgtaattg actgccactt ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg
     3721 cagctgattg cgcatggtgc cgcaaccgtg cggcacccct accgcatgga gataagcatg
     3781 gccacgcagt ccagagaaat cggcattcaa gccaagaaca agcccggtca ctgggtgcaa
     3841 acggaacgca aagcgcatga ggcgtgggcc gggcttattg cgaggaaacc cacggcggca
     3901 atgctgctgc atcacctcgt ggcgcagatg ggccaccaga acgccgtggt ggtcagccag
     3961 aagacacttt ccaagctcat cggacgttct ttgcggacgg tccaatacgc agtcaaggac
     4021 ttggtggccg agcgctggat ctccgtcgtg aagctcaacg gccccggcac cgtgtcggcc
     4081 tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg accagttgcg cctgtcggtg
     4141 ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg aatcgctgtt ggggcatggc
     4201 gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc aactaccgac cggccccggc
     4261 gaggagccgc ccagccagcc cggcattccg ggcatggaac cagacctgcc agccttgacc
     4321 gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc cgatgcccga tgagccgtgt
     4381 tttctggacg atggcgagcc gttggagccg ccgacacggg tcacgctgcc gcgccggtag
     4441 cacttgggtt gcgcagcaac ccgtaagtgc gctgttccag actatcggct gtagccgcct
     4501 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     4561 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccgtttt tatcaggctc
     4621 tgggaggcag aataaatgat catatcgtca attattacct ccacggggag agcctgagca
     4681 aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     4741 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     4801 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     4861 accaggcgtt taagtcgacc aataacatcg cttaggccac acgttcaagt gcagccacag
     4921 gataaatttg cactgagcct gggtgggatt cggactcgac cgcatagcct tcaggagtga
     4981 gttttgtgca ataccaaccg acgacttgac cctgccaagc ggcaccagat ttcttgcgta
     5041 cgcgatcccc taagccaaag gtggcactca ggggaagcgc aaactgccct gcaacgggag
     5101 cgttggcttc atcgctactt tgacccatgt cgaatccttc ttgtgaatct attatggcga
     5161 caagcaaatc gagctctgac tgcctacccc acaacaacta tcagaaagca ccagcacaac
     5221 ggctgcctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata
     5281 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     5341 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     5401 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     5461 gaaccgaaca ggcttatgtc aactgggttc gtgcgagctc atcgatttcg ttccactgag
     5521 cgtcagaccc cgtagatata cagctagccg aattgctgga agtcgtcgcc gccgactaag
     5581 cgctcagcga gcgcctggcc aggtcggcag gggtacgaaa catcgcaacc tgttgactcc
     5641 cttagtaatt tcggaatatt atgctaggtt ccggttcttg gaatgcctgc gcgcgaagtc
     5701 tattcgtgcg ccgactaccg gtccggaagt tcgatgcgca atcgtttctg ctttgcccgg
     5761 aaacagatgc gtcgattttg ttcaatttca ggaataaaca atgccaatca tcaatcaact
     5821 ggttcgcaaa ggccgcgcgt cggaaacgac gaagagcaag agcccggctt tgcaggactg
     5881 cccccagcgt cgcggcgtgt gcacccgtgt gtacaccacg acgccgaaga agcctaactc
     5941 ggcactgcgt aaggttgcca aggttcgtct gacgaacggc ttcgaagtga tttcttacat
     6001 cggtggtgaa ggccacaatc tgcaggaaca ctcggtcgtg ctgattcgcg gcggtcgtgt
     6061 caaggacttg ccgggtgtgc gttaccacat ggttcgtggc tcgctggata cccagggcgt
     6121 caaggatcgt aagcaggctc gctcgaagta cggtgcgaag cgtgcaaaag ctggtaaata
//
