pCMV-dR8.2 dvpr vector (V012794)

Price Information

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V012794 pCMV-dR8.2 dvpr In stock, instant shipping

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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

pCMV-dR8.2 dvpr13380 bp600120018002400300036004200480054006000660072007800840090009600102001080011400120001260013200CMV enhancerCMV promoterHIV-1 polProtein TatProtein VpuRREgp41 peptideNESProtein NefSP6 promoterpBRrevBamAmpR promoterAmpRoriSV40 promotersmall t intronSV40 NLS

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"