Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000941 | pCMV delta R8.2 | 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:
- pCMV delta R8.2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13454 bp
- Type:
- Mammalian Expression, Lentiviral ; Packaging
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV forward
pCMV delta R8.2 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 delta R8.2 vector Sequence
LOCUS V000941 13454 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000941
VERSION V000941
KEYWORDS pCMV delta R8.2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13454)
TITLE Trono Lab Packaging and Envelope Plasmids
REFERENCE 2 (bases 1 to 13454)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13454)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13454
/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"
primer_bind 581..605
/label="LNCX"
/note="Human CMV promoter, forward primer"
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 5895..6068
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate BH5). Accession#: P04612"
/label="Protein Tat"
CDS 6126..6368
/gene="vpu"
/label="Protein Vpu"
/note="Protein Vpu from Human immunodeficiency virus type 1
group M subtype B (isolate BH10). Accession#: P69699"
misc_feature 6532..6765
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6950..6994
/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 7143..7184
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS 7288..7314
/label="NES"
/note="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
CDS 7560..8177
/gene="nef"
/label="Protein Nef"
/note="Protein Nef from Human immunodeficiency virus type 1
group M subtype B (isolate LW123). Accession#: Q70627"
promoter complement(8551..8569)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(8859..8878)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(9092..9111)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9211..9233
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
promoter 9357..9461
/label="AmpR promoter"
CDS 9462..10319
/label="AmpR"
/note="beta-lactamase"
rep_origin 10493..11081
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11235..11252
/label="L4440"
/note="L4440 vector, forward primer"
promoter 11327..11656
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 12790..12855
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 12985..13005
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"