pLP1 vector (V010444)

Price Information

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.

pLP1 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.
• Zhang H, Zhu WQ, Zhao SJ, Li YL, Hou J, Bian CR, Bao CM, Wei Z, Sun J, Li B, Li BA. [Construction and application of lentivirus overexpression vector with two labeling genes fused with CopGFP and PuroR]. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2022 Mar;38(2):108-112. Chinese. doi: 10.12047/j.cjap.6215.2022.028. PMID: 36031566.

pLP1 vector Sequence

LOCUS       Exported                8890 bp DNA     circular SYN 02-SEP-2024
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 8890)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8890)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8890)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8890
                     /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