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pCMV-dR8.91 vector (V007548)

Price Information

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V007548 pCMV-dR8.91 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-dR8.91
Antibiotic Resistance:
Ampicillin
Length:
12051 bp
Type:
Lentiviral vectors
Replication origin:
ori
Promoter:
SV40
Cloning Method:
Enzyme digestion and ligation

pCMV-dR8.91 vector Map

Copy Sequence View Map
pCMV-dR8.9112051 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000CMV enhancerCMV promoterHIV-1 polProtein TatRREgp41 peptideProtein TatNESSP6 promoterAmpR promoterAmpRoriCMV enhancersmall 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-dR8.91 vector Sequence

LOCUS       Exported               12051 bp DNA     circular SYN 02-SEP-2024
DEFINITION  Exported.
ACCESSION   V007548
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12051)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 12051)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 12051)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /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             5255..5428
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate BH5). Accession#: P04612"
     misc_feature    5735..5968
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6153..6197
                     /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             6346..6387
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     CDS             6491..6517
                     /label=NES
                     /note="nuclear export signal from the HIV Rev protein
                     (Fischer et al., 1995)"
     promoter        complement(7140..7158)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7946..8050
                     /label=AmpR promoter
     CDS             8051..8908
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      9082..9670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        9998..10376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          11379..11444
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             11574..11594
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"