Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006935 | pWPI | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pWPI
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11103 bp
- Type:
- Mammalian Expression, Lentiviral, Cre/Lox
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- EF-1α
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- See map
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pWPI vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pWPI vector Sequence
LOCUS V006935 11103 bp DNA circular SYN 20-DEC-2021
DEFINITION Exported.
ACCESSION V006935
VERSION V006935
KEYWORDS pWPI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11103)
TITLE Trono Lab Constitutive Lentiviral Plasmids
REFERENCE 2 (bases 1 to 11103)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11103)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11103
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 182..202
/label="pBABE 3'"
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind complement(272..289)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(443..1031)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1205..2062)
/label="AmpR"
/note="beta-lactamase"
promoter complement(2063..2167)
/label="AmpR promoter"
primer_bind 2235..2253
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2291..2313)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 2413..2432
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2646..2665
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2955..2973
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
LTR complement(2998..3178)
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
protein_bind 3198..3231
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature complement(3375..3963)
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind 3976..3992
/label="pBluescriptKS"
/note="For pBluescript vector"
primer_bind 3977..3993
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(4072..4788)
/label="EGFP"
/note="enhanced GFP"
misc_feature complement(4793..5355)
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature complement(5409..5526)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter complement(5574..6752)
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS complement(6945..6986)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(7135..7179)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(7364..7597)
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature complement(8090..8215)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(8262..8895)
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
polyA_signal complement(9063..9197)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(9622..9642)
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
intron complement(9772..9837)
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
primer_bind complement(11039..11058)
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"