Invitrogen™ pLenti6/V5 Directional TOPO™ Cloning Kit

Description

Directional TOPO Cloning site for rapid and efficient directional cloning of blunt-end PCR products

• Contains TOPO-adapted ViraPower lentiviral expression vector, pLenti6/V5-D-TOPO for quick PCR-based cloning and expression of a target gene in dividing and nondividing mammalian cells
• Vector has CMV promoter for driving high-level, constitutive expression of the target gene and blasticidin selection marker for stable selection in mammalian cells
• High efficiency and rapid cloning
• Produces replication-incompetent virus for enhanced biosafety of system
• Rouse Sarcoma Virus (RSV) enhancer/promoter for Tat-independent production of viral mRNA in the producer cell line
• Modified HIV-1 5' and 3' Long Terminal Repeats (LTR) for viral packaging and reverse transcription
• HIV-1 psi (ψ) packaging sequence for viral packaging
• HIV Rev response element (RRE) for Rev-dependent nuclear export of unspliced viral mRNA
• (CMV) immediate early promoter for high-level constitutive expression of gene of interest in mammalian cells
• C-terminal V5 epitope for detection of recombinant protein
• Blasticidin (bsd) resistance gene for selection in E. coli and mammalian cells
• Ampicillin resistance gene for selection in E. coli
• pUC origin for high-copy replication and maintenance of plasmid in E. coli

Constitutive Expression, Mammalian Expression, Protein Expression, Proteins, Expression, Isolation and Analysis, Viral Gene Delivery

Specifications

• Cloning Method: Directional TOPO™
• Constitutive or Inducible System: Constitutive
• Content And Storage: pLenti⁄V5-D-TOPO™ reagents (Box 1) Store at -20°C
  • pLenti6⁄V5-D-TOPO™ Reagents (15-20 ng⁄μl linearized plasmid DNA in: 50% glycerol, 50 mM Tris-HCl, pH 7.4 (at 25°C),1 mM EDTA, 2 mM DTT, 0.1% Triton X-100, 100 μg⁄ml BSA, 30 μM bromophenol blue)
  • dNTP Mix (12.5 mM each dATP, dCTP, dGTP, dTTP in water, pH8 at a total volume of 10 μl)
  • Salt Solution (1.2 M NaCl and 0.06 M MgCl2, volume of 50 μl)
  • Water (1 ml)
  • CMV Forward Sequencing Primer (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 20 μl)
  • Control PCR Primers (0.1 μg⁄μl each in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
  • Control PCR Template (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
  • pLenti6⁄V5-GW⁄lacZ (lyophilized in TE Buffer, pH 8, 10 μg)
  
  One Shot™ Stbl3™ Chemically Competent E. coli (Box 2) Store at -80°C
  • S.O.C. Medium (2% Trypton, 0.5% Yeast Extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl₂, 10 mM MgSO₄, 20 mM glucose. 6ml) May be stored at +4°C or room temperature
  • Stbl3™ Cells (21 x 50 μl)
  • pUC19 Control DNA (10 pg⁄μl in 5 mM Tris-HCl, 0.5 mM EDTA, pH 8. 50 μl)
• Delivery Type: Lentiviral
• For Use With (Application): Viral Expression
• No. of Reactions: 20 reactions
• Product Line: TOPO™, ViraPower™
• Product Type: TOPO Cloning Kit
• Promoter: CMV
• Protein Tag: V5 Epitope Tag
• Quantity: 20 Reactions
• Selection Agent (Eukaryotic): Blasticidin
• Vector: pLenti, Directional TOPO Vectors

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