Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012390 | pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt) | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7229 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- SFFV
- Cloning Method:
- Restriction Enzyme
pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt) vector 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.
pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt) vector Sequence
LOCUS V012390 7229 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012390
VERSION V012390
KEYWORDS pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7229)
AUTHORS Certo MT, Ryu BY, Annis JE, Garibov M, Jarjour J, Rawlings DJ,
Scharenberg AM
TITLE Tracking genome engineering outcome at individual DNA breakpoints.
JOURNAL Nat Methods. 2011 Jul 10;8(8):671-6. doi: 10.1038/nmeth.1648.
PUBMED 21743461
REFERENCE 2 (bases 1 to 7229)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7229)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth";
journalName: "Nat Methods"; date: "2011-07-10- 10"; volume: "8";
issue: "8"; pages: "671-6"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7229
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 9..235
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 236..416
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 463..588
/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 1955..2361
/label="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 2376..2385
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2388..3056
/label="(3-F)Tyr-EGFP"
/note="(3-F)Tyr-EGFP is a basic (constitutively
fluorescent) green fluorescent protein published in 2004,
derived from Aequorea victoria."
promoter 3086..3297
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 3306..3315
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3321..3347
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 3357..3377
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 3375..4079
/gene="SCEI"
/label="Intron-encoded endonuclease I-SceI"
/note="Intron-encoded endonuclease I-SceI from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P03882"
misc_feature 4104..4692
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
protein_bind complement(4768..4801)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
polyA_signal 5012..5236
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5309..5326)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(5480..6068)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6242..7099)
/label="AmpR"
/note="beta-lactamase"
promoter complement(7100..7204)
/label="AmpR promoter"