LOCUS       Exported                6722 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBADSce, complete sequence.
ACCESSION   FJ797515
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6722)
  AUTHORS   Lopez CM, Rholl DA, Trunck LA, Schweizer HP.
  TITLE     Versatile dual-technology system for markerless allele replacement 
            in Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 75 (20), 6496-6503 (2009)
  PUBMED    19700544
REFERENCE   2  (bases 1 to 6722)
  AUTHORS   Lopez CM, Schweizer HP.
  TITLE     Homing Endonuclease-based System for Select Agent-compliant 
            Markerless Gene Replacement in Burkholderia pseudomallei
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6722)
  AUTHORS   Lopez CM, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2009) Microbiology, Immunology and Patholofy, 
            Colorado State University, 3205 Rampart Rd., Fort Collins, CO 80523,
            USA
REFERENCE   4  (bases 1 to 6722)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6722
                     /organism="Expression vector pBADSce"
                     /mol_type="other DNA"
                     /label=pBADSce
                     /note="pBADSce"
                     /db_xref="taxon:629243"
     regulatory      14..439
                     /regulatory_class="terminator"
                     /note="Escherichia coli rrnB fragment containing 
                     terminators T1 and T2"
     terminator      216..302
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      394..421
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        440..531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(930..1304)
                     /codon_start=1
                     /gene="ble"
                     /product="Ble"
                     /label=ble
                     /note="zeocin resistance protein"
                     /protein_id="ACO48885.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            complement(930..1304)
                     /gene="ble"
                     /label=ble
     regulatory      complement(1305..1371)
                     /regulatory_class="promoter"
                     /note="pEM7"
     promoter        complement(1323..1370)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      1490..1841
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             1855..2688
                     /codon_start=1
                     /gene="rep-ts"
                     /product="Rep-TS"
                     /label=rep-ts
                     /note="TS replication protein"
                     /protein_id="ACO48886.1"
                     /translation="MASPPMVYKSNALVEAAYCLSVQEQRIVLASISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMGSTHAIRLYELLTQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKARKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            1855..2688
                     /gene="rep-ts"
                     /label=rep-ts
     rep_origin      2915..3323
                     /label=M13 replication origin
                     /note="M13 replication origin"
     rep_origin      2918..3373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3379..4076
                     /label=pBBR22 replication origin
                     /note="pBBR22 replication origin"
     rep_origin      3484..4072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4769..5647)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="arabinose operon subunit"
                     /protein_id="ACO48887.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(4769..5647)
                     /gene="araC"
                     /label=araC
     promoter        5674..5958
                     /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      5923..5950
                     /regulatory_class="promoter"
                     /note="pBAD"
     CDS             5993..6706
                     /codon_start=1
                     /gene="I-Sce I"
                     /product="I-Sce I"
                     /label=I-Sce I
                     /note="homing endonuclease"
                     /protein_id="ACO48888.1"
                     /translation="MHMKNIKKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILG
                     DAYIRSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKERVNHLGNLVITWGAQ
                     TFKHQAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIV
                     LNTQSFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQM
                     MYKLPNTISSETFLK"
     gene            5993..6706
                     /gene="I-Sce I"
                     /label=I-Sce I