LOCUS       Exported                3728 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  FRT-Tp-FRT expression vector pwFRT-PCS12-Tp, complete sequence.
ACCESSION   EU334849
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3728)
  AUTHORS   Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.
  TITLE     Genetic tools for allelic replacement in Burkholderia species
  JOURNAL   Appl. Environ. Microbiol. 74 (14), 4498-4508 (2008)
  PUBMED    18502918
REFERENCE   2  (bases 1 to 3728)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2007) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 3728)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3728
                     /organism="FRT-Tp-FRT expression vector pwFRT-PCS12-Tp"
                     /mol_type="other DNA"
                     /db_xref="taxon:491856"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="ABY61790.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1268..1884
                     /label=ColE1 origin
                     /note="ColE1 origin"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     2289..2362
                     /note="FRT; Flp-recombination-target"
     protein_bind    complement(2306..2353)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2393..2434
                     /regulatory_class="enhancer"
                     /note="Burkholderia cenocepacia S12 promoter"
     RBS             2445..2467
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        2565..2593
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2920..3156
                     /codon_start=1
                     /gene="dhfr"
                     /product="DhfrII"
                     /label=dhfr
                     /note="dihydrofolate reductase protein"
                     /protein_id="ABY61791.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     gene            2920..3156
                     /gene="dhfr"
                     /label=dhfr
     misc_recomb     3203..3276
                     /note="FRT; Flp-recombination-target"
     protein_bind    complement(3220..3267)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(3334..3350)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"