Invitrogen™ ViraPower™ HiPerform™ Promoterless Gateway™ Vector Kit

Invitrogen™ ViraPower™ HiPerform™ Promoterless Gateway™ Vector Kit

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
  
  

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
  
  

Contents

• pLenti6.4⁄R4R2⁄V5-DEST™ Kit which includes:
• 6 µg of pLenti6.4⁄R4R2⁄V5-DEST™ vector (40 µL of vector at 150 ng⁄µL in TE Buffer, pH 8.0)
• 10 µg of pLenti6.4⁄CMV⁄V5-MSGW⁄lacZ plasmid (20 µL of vector at 0.5 µg⁄µL in TE Buffer, pH 8.0)
• 10 µg of pENTR 5’⁄CMVp plasmid (20 µL of plasmid at 0.5 µg⁄µL in TE Buffer, pH 8.0)
• 10 µg of pENTR 5’⁄EF1αp plasmid (20 µL of plasmid at 0.5 µg⁄µL in TE Buffer, pH 8.0)
• pENTR 5’ TOPO™ TA Cloning Kit which includes: K95120
• pENTR 5’ TOPO™ Reagents
• One Shot™ TOP10 Chemically Competent E.coli: C404003
• One Shot™ Stbl3™ Chemically Competent E. coli: 21 X 50 µL of Stbl3™ Cells, 6 mL of S.O.C. Medium and 50 µL of pUC19 control DNA (10 pg⁄µL): C737303
• pENTR™ Gus positive control plasmid