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pwFRT-PCS12-Tp vector (V002138#)

Basic Vector Information

Basic Vector Information
Vector Name pwFRT-PCS12-Tp Antibiotic Resistance Ampicillin
Length 3728 bp Type FRT-Tp-FRT expression vector
Source Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.

pwFRT-PCS12-Tp vector Vector Map

pwFRT-PCS12-Tp3728 bp60012001800240030003600AmpR promoterblaoriCAP binding sitelac promoterlac operatorM13 revFRTBurkholderia cenocepacia S12 promoterRBSPc promoterdhfrFRTM13 fwd

Plasmid Resuspension protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pwFRT-PCS12-Tp vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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"

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