Invitrogen™ ViraPower™ HiPerform™ Promoterless Gateway™ Vector Kit

Description

The ViraPower HiPerform Promoterless Gateway Vector Kit contains MultiSite Gateway-adapted ViraPower HiPerform promoterless lentiviral expression vector, pLenti6.4/R4R2/V5-DEST for easy recombination-based cloning and expression of a target gene in dividing and non-dividing mammalian cells.

The ViraPower™ HiPerform™ Promoterless Gateway™ Vector Kit contains the MultiSite Gateway™-adapted ViraPower™ HiPerform™ promoterless lentiviral expression vector, pLenti6.4/R4R2/V5-DEST™ for easy recombination-based cloning and lentiviral-based high-level expression of a target gene from any promoter of choice in dividing and non-dividing mammalian cells. The pLenti6.4/R4R2/V5-DEST™ vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements.

Advantages

• Generates replication-incompetent lentivirus for transducing dividing and non-dividing mammalian cells
• Easy, simultaneous, recombination-based cloning of multiple DNA fragments in a defined order and orientation using MultiSite Gateway™ technology
• Expression of the target gene under the control of a promoter of choice
• Stable, long-term expression
• Enhanced protein expression, up to 4-fold or greater, compared to traditional lentiviral expression systems

Key Features

• WPRE from the woodchuck hepatitis virus, increases transgene expression and cPPT from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome, thus increasing viral titer. WPRE and cPPT together produce at least a four-fold increase in protein expression in most cell types, compared to other vectors that do not contain these elements.
• Promoterless vector to express the target gene under the control of a promoter of choice
• Blasticidin selection marker for stable selection under control of PGK promoter for long-term, persistent expression

Kit includes

• pLenti6.4/R4R2/V5-DEST™ Kit
• pENTR™ 5' TOPO™ TA Cloning Kit (Cat. No. K59120)
• One Shot™ Stbl3™ Chemically Competent E. coli (Cat. No. C737303)
• pENTR™ Gus positive control plasmid

Specifications

• Constitutive or Inducible System: Constitutive
• Delivery Type: Lentiviral
• Promoter: None (Promoterless), EF-1α, CMV
• Product Type: Vector Kit
• Selection Agent (Eukaryotic): Blasticidin
• Content And Storage:

  pLenti6.4/R4R2/V5-DEST™ Kit
  • pLenti6.4/R4R2/V5-DEST™ vector (40 μL of vector at 150 ng/μL in TE Buffer, pH 8.0), 6 μg
  • pLenti6.4/CMV/V5-MSGW/lacZ plasmid (20 μL of vector at 0.5 μg/μL in TE Buffer, pH 8.0), 10 μg
  • pENTR 5'/CMVp plasmid (20 μL of plasmid at 0.5 μg/μL in TE Buffer, pH 8.0), 10 μg
  • pENTR 5'/EF1αp plasmid (20 μL of plasmid at 0.5 μg/μL in TE Buffer, pH 8.0), 10 μg
  Store at -20°C
  
  pENTR 5' TOPO™ TA Cloning Kit (Cat. No. K95120)
  • pENTR 5' TOPO™ Reagents, store at -20°C.
  • One Shot™ TOP10 Chemically Competent E.coli (Cat. No. C404003), store at -80°C.
  
  One Shot™ Stbl3™ Chemically Competent E. coli (Cat. No. C737303)
  • Stbl3™ Cells, 6 mL of S.O.C. Medium, 21 x 50 μL
  • pUC19 control DNA (10 pg/μL), 50 μL
  Store at -80°C
  
  pENTR™ Gus positive control plasmid, store at -20°C

• Antibiotic Resistance Bacterial: Ampicillin (AmpR), Blasticidin (BsdR)
• Cleavage: No Cleavage Site
• Gateway Enzyme: LR Clonase™ II Plus, LR Clonase™ II
• PCR Enzyme: Proof-Reading Polymerase
• Protein Tag Position (to your gene): C-terminal
• Replication Origin: pUC Origin
• Selection Marker Promoter: PGK Promoter
• Selection Marker Eukaryotic: BsdR
• Special Elements: WPRE, cPPT
• Protein Tag: V5 Epitope Tag
• Cloning Method: Gateway
• Quantity: 20 Reactions
• Vector: pLenti
• Product Line: Gateway™, HiPerform™, ViraPower™
• For Use With (Application): Clone your own Promoter, Viral Expression

For Research Use Only. Not for use in diagnostic procedures.