ViraPower™ HiPerform™ T-REx™ Gateway® Vector Kit

Catalog number: A11144

Invitrogen™  Related applications: Protein Expression

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Description

The ViraPower™ HiPerform™ T-REx™ Gateway® Vector Kit contains the Gateway®-adapted ViraPower™ HiPerform™ tetracycline-regulated lentiviral expression vector, pLenti6.3⁄TO⁄V5-DEST™ for easy recombination-based cloning and lentiviral-based, regulated, high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti6.3⁄ TO⁄ 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 recombination-based cloning using Gateway® technology
• Stable, long-term, tetracycline-regulated 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.
• Hybrid promoter consisting of the human cytomegalovirus (CMV) promoter and two tetracycline operator 2 (TetO2) sites for high-level, regulated expression of the target gene
• Blasticidin selection marker for stable selection under control of SV40 promoter

Kit includes
• pLenti6.3⁄TO⁄V5-DEST™ vector
• pLenti3.3⁄TR vector
• pLenti6.3⁄TO⁄V5-GW⁄lacZ plasmid
• Tetracycline
• One Shot® Stbl3™ Chemically Competent E. coli (Cat # C737303)
• pENTRTM Gus positive control plasmid.

For research use only. Not intended for any therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: CMV⁄TO
Key Function: Viral Expression
Protein Tag or Fusion: V5 Epitope Tag
Delivery Method: Lentiviral
Cloning Method: Gateway®
Selection Agent (Eukaryotic): Blasticidin
Vector Type: pLenti
Cleavage: No Cleavage Site
PCR Enzyme: Proof-Reading Polymerase
Product Size: 20 reactions
Gateway Enzyme: LR Clonase® II, LR Clonase® II Plus
Inducing Agent: Tetracycline, Doxycycline
Special Elements: WPRE, cPPT
Replication Origin: pUC Origin
Selection Marker Promoter: PGK Promoter
Selection Marker (Eukaryotic): BsdR
Constitutive or Inducible System: Inducible
Antibiotic Resistance (Bacterial): Ampicillin (AmpR), Blasticidin (BsdR)
Protein Tag Position (to your gene): C-terminal

Contents & storage

Contents
The ViraPower™ HiPerform™ T-REx™ Gateway® Vector Kit includes the following components:
• 6 ug of pLenti6.3⁄TO⁄V5-DEST™ vector (40 µL of vector at 150 ng⁄µL in TE Buffer, pH 8.0)
• 20 ug of pLenti3.3⁄TR vector (40 µL of vector at 0.5 µg⁄µL in TE Buffer, pH 8.0)
• 10 ug of pLenti6.3⁄TO⁄V5-GW⁄lacZ plasmid (20 µL of plasmid at 0.5 µg⁄µL in TE Buffer, pH 8.0)
• 1 ml of Tetracycline (10 mg⁄mL in water)
• One Shot™ Stbl3™ Chemically Competent E. coli (Cat # C737303): 21 X 50 µL of Stbl3™ Cells, 6 mL of S.O.C. Medium and 50 µL of pUC19 control DNA (10 pg⁄µL).
• pENTR™ Gus positive control plasmid.

Storage
• Vectors: -20°C
• Tetracycline: -20°C (protected from light)
• One Shot™ Stbl3™ Chemically Competent E. coli: -80°C
• pENTR™ Gus positive Control: -20°C

Documents

Manuals & protocols