LOCUS       Exported                6775 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector pMLBAD, complete sequence.
ACCESSION   AY112733
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. 68 (12), 5956-5964 (2002)
  PUBMED    12450816
REFERENCE   2  (bases 1 to 6775)
  AUTHORS   Lefebre MD, Valvano MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-MAY-2002) Microbiology and Immunology, University of 
            Western Ontario, Dental Sciences Building Rm 3000, London, ONT N6A 
            5C1, Canada
REFERENCE   3  (bases 1 to 6775)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6775
                     /organism="Broad host range vector pMLBAD"
                     /host="Burkholderia cepacia"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:198728"
     promoter        55..83
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             410..646
                     /codon_start=1
                     /gene="dhfrII"
                     /product="dihydrofolate reductase"
                     /label=dhfrII
                     /note="trimethoprim resistance determinant"
                     /protein_id="AAM63381.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     gene            410..646
                     /gene="dhfrII"
                     /label=dhfrII
     CDS             complement(869..1798)
                     /codon_start=1
                     /gene="araC"
                     /product="transcriptional regulator AraC"
                     /label=araC
                     /protein_id="AAM63382.1"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     gene            complement(869..1798)
                     /gene="araC"
                     /label=araC
     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)"
     regulatory      2023..2050
                     /regulatory_class="promoter"
                     /note="P-BAD"
     misc_feature    2084..2139
                     /note="multiple cloning site; EcoRI to HindIII"
     regulatory      2140..2565
                     /regulatory_class="terminator"
                     /note="rrnB"
     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"
     promoter        complement(3402..3504)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(3654..4673)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /function="required for plasmid mobilization"
                     /label=mob
                     /protein_id="AAM63383.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS"
     gene            complement(3654..4673)
                     /gene="mob"
                     /label=mob
     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"
     rep_origin      5469..5480
     CDS             5667..6329
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /function="required for plasmid replication"
                     /label=rep
                     /protein_id="AAM63384.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            5667..6329
                     /gene="rep"
                     /label=rep