Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V012790 | pRSV-Rev | 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
3rd generation lentiviral packaging plasmid; Contains Rev; also requires pMDLg/pRRE and envelope expressing plasmid.
- Vector Name:
- pRSV-Rev
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4180 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Didier Trono
- Copy Number:
- High copy number
- Promoter:
- RSV
- Growth Strain(s):
- JM108
- Growth Temperature:
- 37℃
pRSV-Rev 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
- Farzaneh M, Sayyah M, Eshraghi HR, Panahi N, Mirzapourdelavar H, Gholami Pourbadie H. The Lentiviral Vector Pseudotyped by Modified Rabies Glycoprotein Does Not Cause Reactive Gliosis and Neurodegeneration in Rat Hippocampus. Iran Biomed J. 2019 Sep;23(5):324-9. doi: 10.29252/.23.5.324. Epub 2019 May 19. PMID: 31103020; PMCID: PMC6661131.
pRSV-Rev vector Sequence
LOCUS Exported 4180 bp DNA circular SYN 29-AUG-2024 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4180) TITLE Direct Submission REFERENCE 2 (bases 1 to 4180) TITLE Direct Submission REFERENCE 3 (bases 1 to 4180) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..4180 /mol_type="other DNA" /organism="synthetic DNA construct" source 1208..1213 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 107..128 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 366..627 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" CDS 756..1103 /codon_start=1 /label=Rev /note="human immunodeficiency virus 1 (HIV-1) Rev protein" /translation="MAGRSGDSDEDLLKAVRLIKFLYQSNPPPNPEGTRQARRNRRRRW RERQRQIHSISERILSTYLGRSAEPVPLQLPPLERLTLDCNEDCGTSGTQGVGSPQILV ESPTILESGAKE" primer_bind complement(1190..1206) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 1538..1993 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2275..2379 /label=AmpR promoter CDS 2380..3237 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3411..3999 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"