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pCMV-VSV-G vector (V010488)

Price Information

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V010488 pCMV-VSV-G In stock, instant shipping

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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

Copy Sequence View Map
pCMV-VSV-G6507 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300CMV enhancerCMV promoterbeta-globin intronVSV-Gbeta-globin poly(A) signalM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterf1 oriM13 fwdT7 promoter

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"