LOCUS       Exported                6601 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pGPI-SceI-XCm, complete sequence.
ACCESSION   KC409641
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  AUTHORS   Hamad MA, Skeldon AM, Valvano MA.
  TITLE     Construction of aminoglycoside-sensitive Burkholderia cenocepacia 
            strains for use in studies of intracellular bacteria with the 
            gentamicin protection assay
  JOURNAL   Appl. Environ. Microbiol. 76 (10), 3170-3176 (2010)
  PUBMED    20348312
REFERENCE   2  (bases 1 to 6601)
  AUTHORS   Hamad MA, Skeldon AM, Valvano MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-DEC-2012) Microbiology and Immunology, Western 
            University, 1151 Richmond Street North, London, Ontario N6A 5C1, 
            Canada
REFERENCE   3  (bases 1 to 6601)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6601
                     /organism="Suicide vector pGPI-SceI-XCm"
                     /mol_type="other DNA"
                     /db_xref="taxon:1276082"
     oriT            626..735
                     /note="incP origin of transfer"
     CDS             768..1139
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             873..1139
                     /codon_start=1
                     /product="TraJ"
                     /label=TraJ
                     /protein_id="AGC79934.1"
                     /translation="MSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLW
                     LTDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP"
     CDS             complement(1102..1527)
                     /codon_start=1
                     /product="TraI"
                     /label=TraI
                     /protein_id="AGC79933.1"
                     /translation="MNVQVVGVVMHGTDALMLGEAKPSADAVLNRAQRLRVGLLSRAEA
                     NQQVIGLVGLGTGVAVLGRHHLGHDSGQGVCLAVRDAHVTQALGLALLVGDVLHQLREV
                     ALLDGAHGDVLGNHAHPPAVLAAKKVMALPSGGPRPS"
     rep_origin      1757..2148
     rep_origin      1757..2145
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    2150..2178
                     /label=multiple cloning site 1
                     /note="multiple cloning site 1"
     promoter        2194..2222
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             2254..3177
                     /codon_start=1
                     /product="XylE reporter protein"
                     /label=XylE reporter protein
                     /protein_id="AGC79932.1"
                     /translation="MNKGVMRPGHVQLRVLDMSKALEHYVELLGLIEMDRDDQGRVYLK
                     AWTEVDKFSLVLREADEPGMDFMGFKVVDEDALRQLERDLMAYGCAVEQLPAGELNSCG
                     RRVRFQAPSGHHFELYADKEYTGKWGLNDVNPEAWPRDLKGMAAVRFDHALMYGDELPA
                     TYDLFTKVLGFYLAEQVLDENGTRVAQFLSLSTKAHDVAFIHHPEKGRLHHVSFHLETW
                     EDLLRAADLISMTDTSIDIGPTRHGLTHGKTIYFFDPSGNRNEVFCGGDYNYPDHKPVT
                     WTTDQLGKAIFYHDRILNERFMTVLT"
     misc_feature    3183..3216
                     /label=multiple cloning site 2
                     /note="multiple cloning site 2"
     promoter        3945..3973
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4299..4535
                     /codon_start=1
                     /product="DHFR trimethoprim resistance protein"
                     /label=DHFR trimethoprim resistance protein
                     /protein_id="AGC79931.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     misc_feature    4538..4555
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"
     promoter        4927..5029
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5030..5668
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label=chloramphenicol acetyltransferase
                     /protein_id="AGC79930.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"