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
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
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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