Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012169 | pRRLSIN.cPPT.PGK-GFP.WPRE | 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
The pRRLSIN.cPPT.PGK-GFP.WPRE is a 3rd generation lentiviral backbone from Didier Trono Lab.
- Vector Name:
- pRRLSIN.cPPT.PGK-GFP.WPRE
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7388 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- hPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- See map
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pRRLSIN.cPPT.PGK-GFP.WPRE 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.
References
- Trono Lab Misc Plasmids
pRRLSIN.cPPT.PGK-GFP.WPRE vector Sequence
LOCUS V012169 7388 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012169
VERSION V012169
KEYWORDS pRRLSIN.cPPT.PGK-GFP.WPRE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7388)
TITLE Trono Lab Misc Plasmids
REFERENCE 2 (bases 1 to 7388)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7388)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7388
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 8..234
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 235..415
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 462..587
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1080..1313
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1498..1542
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1691..1732
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 1809..1926
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1972..2482
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
CDS 2504..3220
/label="EGFP"
/note="enhanced GFP"
misc_feature 3239..3827
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3914..4147
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4219..4353
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4380..4515
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(4536..4554)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4564..4580)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4722..5177
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5203..5307
/label="AmpR promoter"
CDS 5308..6165
/label="AmpR"
/note="beta-lactamase"
rep_origin 6339..6927
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7081..7098
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 7215..7236
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7251..7281
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 7289..7305
/label="lac operator"
/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 7313..7329
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 7350..7368
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"