Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012794 | pCMV-dR8.2 dvpr | 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
2nd generation lentiviral packaging plasmid. Can be used with 2nd and 3rd generation transfer vectors. Use in conjunction with an envelope plasmid such as pCMV-VSV-G.
- Vector Name:
- pCMV-dR8.2 dvpr
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13380 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Bob Weinberg
- Copy Number:
- High copy number
- Promoter:
- CMV
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pCMV-dR8.2 dvpr 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.
pCMV-dR8.2 dvpr vector Sequence
LOCUS V012794 13380 bp DNA circular SYN 12-MAY-2024
DEFINITION Exported.
ACCESSION V012794
VERSION V012794
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13380)
AUTHORS Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS,
Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD
TITLE Lentivirus-delivered stable gene silencing by RNAi in primary cells.
JOURNAL RNA 2003 Apr;9(4):493-501.
PUBMED 12649500
REFERENCE 2 (bases 1 to 13380)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13380)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13380)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "RNA"; date:
"2003-04"; volume: "9(4)"; pages: "493-501"
SGRef: number: 2; type: "Journal Article"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13380
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 855..2354
/label="HIV-1 gag"
/note="gag protein from human immunodeficiency virus 1"
CDS 2150..5158
/label="HIV-1 pol"
/note="pol protein from human immunodeficiency virus 1"
CDS 5106..5681
/gene="vif"
/label="Virion infectivity factor"
/note="Virion infectivity factor from Human
immunodeficiency virus type 1 group M subtype B (isolate
NY5). Accession#: P12504"
CDS 5821..5994
/label="Protein Tat"
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate BH5). Accession#: P04612"
CDS 6052..6294
/gene="vpu"
/label="Protein Vpu"
/note="Protein Vpu from Human immunodeficiency virus type 1
group M subtype B (isolate BH10). Accession#: P69699"
misc_feature 6458..6691
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6876..6920
/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 7214..7240
/label="NES"
/note="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
CDS 7486..8103
/gene="nef"
/label="Protein Nef"
/note="Protein Nef from Human immunodeficiency virus type 1
group M subtype B (isolate LW123). Accession#: Q70627"
promoter complement(8477..8495)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(8785..8804)
/label="pBRrevBam"
/note="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 9283..9387
/label="AmpR promoter"
CDS 9388..10245
/label="AmpR"
/note="beta-lactamase"
rep_origin 10419..11007
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 11253..11582
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 12716..12781
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 12911..12931
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"