Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V010488 | pCMV-VSV-G | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pCMV-VSV-G
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6507 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Yee JK, Miyanohara A, LaPorte P, Bouic K, Burns JC, Friedmann T.
- Copy Number:
- High copy number
pCMV-VSV-G 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
- Yan YF, Wang ZY, Pu SS, Wen HL, Huang T, Song YY, Xu HZ, Zhao L. HIV-1B gp120 genes from one patient with AIDS dementia complex can affect the secretion of tumor necrosis factor and interleukin 1β in glial cells. Chin Med J (Engl). 2011 Dec;124(24):4217-22. PMID: 22340390.
- Zhao L, Pu SS, Gao WH, Chi YY, Wen HL, Wang ZY, Song YY, Yu XJ. Effects of HIV-1 tat on secretion of TNF-α and IL-1β by U87 cells in AIDS patients with or without AIDS dementia complex. Biomed Environ Sci. 2014 Feb;27(2):111-7. doi: 10.3967/bes2014.024. PMID: 24625401.
- Icsel C, Yilmaz VT. DNA binding and cleavage studies of two palladium(II) saccharinate complexes with terpyridine. DNA Cell Biol. 2013 Apr;32(4):165-72. doi: 10.1089/dna.2012.1959. Epub 2013 Mar 8. PMID: 23473538.
pCMV-VSV-G vector Sequence
LOCUS 40924_12005 6507 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector for constitutive expression of VSV-G glycoprotein. Also known
as pHCMV-G or pHCMV-VSV-G.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6507)
AUTHORS Yee JK, Miyanohara A, LaPorte P, Bouic K, Burns JC, Friedmann T.
TITLE A general method for the generation of high-titer, pantropic
retroviral vectors: highly efficient infection of primary
hepatocytes.
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 1994;91:9564-8.
PUBMED 7937806
REFERENCE 2 (bases 1 to 6507)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6507)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
Acad. Sci. U.S.A."; date: "1994"; volume: "91"; pages: "9564-8"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT The sequence from GenBank accession number AJ318514 was edited to
match data associated with Addgene plasmid number 8454.
FEATURES Location/Qualifiers
source 1..6507
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 83..462
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 463..666
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 778..1350
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
CDS 1436..2968
/codon_start=1
/label=VSV-G
/note="vesicular stomatitis virus G glycoprotein"
/translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
polyA_signal 3263..3318
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3676..3692)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3700..3716)
/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)."
promoter complement(3724..3754)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3769..3790)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4078..4666)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4840..5697)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5698..5802)
/label=AmpR promoter
rep_origin complement(5828..6283)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6425..6441
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 6451..6469
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"