Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010444 | pLP1 | 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
pLP1 is a lentiviral packaging plasmid containing the HIV-1 gag and pol genes.
- Vector Name:
- pLP1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8889 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- Promoter:
- CMV+intron
- Growth Strain(s):
- Stbl3
pLP1 vector 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 HK, Yuan ZG, Song HQ, Petersen E, Zhou Y, Ren D, Zhou DH, Li HX, Lin RQ, Yang GL, Zhu XQ. Vaccination with a DNA vaccine coding for perforin-like protein 1 and MIC6 induces significant protective immunity against Toxoplasma gondii. Clin Vaccine Immunol. 2012 May;19(5):684-9.
pLP1 vector Sequence
LOCUS pLP1. 8889 bp DNA circular SYN 01-JAN-1980
DEFINITION Lentiviral packaging plasmid containing the HIV-1 gag and pol genes.
ACCESSION .
VERSION .
KEYWORDS pLP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8889)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8889)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8889
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 79..458
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 459..662
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 796..1271
/label=beta-globin intron
/note="internally truncated intron from human beta-globin"
CDS 1355..2854
/label=HIV-1 gag
/note="gag protein from human immunodeficiency virus 1"
CDS 2650..5658
/label=HIV-1 pol
/note="pol protein from human immunodeficiency virus 1"
misc_feature 5686..5919
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
polyA_signal 6150..6544
/label=beta-globin poly(A)
/note="human beta-globin polyadenylation signal"
rep_origin complement(7054..7642)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7816..8673)
/label=AmpR
/note="beta-lactamase"
promoter complement(8674..8778)
/label=AmpR promoter