pLVX-Puro vector (V010426)

Price Information

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V010426 pLVX-Puro In stock, instant shipping

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

The pLVX-Puro is a lentiviral expression vector with the constitutive CMV promoter and a puromycin resistance marker.

Vector Name:
pLVX-Puro
Antibiotic Resistance:
Ampicillin
Length:
8055 bp
Type:
Viral Expression & Packaging Vectors
Replication origin:
ori
Source/Author:
Clontech
Copy Number:
High copy number
Promoter:
mPGK

pLVX-Puro vector Map

pLVX-Puro8055 bp4008001200160020002400280032003600400044004800520056006000640068007200760080003' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSCMV enhancerCMV promoterMCSPGK promoterPuroRWPRE5' LTRM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterSV40 poly(A) signal

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

  • Li X, Chen Z, Li X, Wang H. In vitro analysis of the role of tumor necrosis factor‑stimulated gene‑6 in keloid. Mol Med Rep. 2019 Feb;19(2):919-926. doi: 10.3892/mmr.2018.9767. Epub 2018 Dec 14. PMID: 30569148; PMCID: PMC6323212.
  • Abbasi Pashaki P, Rahim F, Habibi Roudkenar M, Razavi-Toosi S, Ebrahimi A. MicroRNA Tough Decoy Knockdowns miR-195 and Represses Hypertrophy in Chondrocytes. Appl Biochem Biotechnol. 2020 Jul;191(3):1056-1071. doi: 10.1007/s12010-020-03229-6. Epub 2020 Jan 20. PMID: 31956957.
  • Han X, Shi F, Guo S, Li Y, Wang H, Song C, Wu S. LINC02086 promotes cell viability and inhibits cell apoptosis in breast cancer by sponging miR-6757-5p and up-regulating EPHA2. World J Surg Oncol. 2023 Nov 27;21(1):371. doi: 10.1186/s12957-023-03245-w. PMID: 38008720; PMCID: PMC10680215.

pLVX-Puro vector Sequence

LOCUS       Exported                8055 bp DNA     circular SYN 02-SEP-2024
DEFINITION  Exported.
ACCESSION   V010426
VERSION     .
KEYWORDS    pLVX-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8055)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8055)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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             1914..1955
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2027..2144
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2201..2504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    2802..2867
                     /label=MCS
                     /note="multiple cloning site"
     promoter        2890..3389
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3410..4006
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    4023..4611
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4818..5451
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5580..5596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5604..5620)
                     /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(5628..5658)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5673..5694)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5982..6570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6744..7601)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7602..7706)
                     /label=AmpR promoter
     polyA_signal    7754..7888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"