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Invitrogen™ pLenti6/V5 Directional TOPO™ Cloning Kit 
Directional TOPO Cloning site for rapid and efficient directional cloning of blunt-end PCR products
Supplier: Invitrogen™ K495510
Description
- 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
| Directional TOPO™ | |
| 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) |
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| Viral Expression | |
| TOPO, ViraPower | |
| CMV | |
| 20 Reactions | |
| pLenti, Directional TOPO Vectors |
| Constitutive | |
| Lentiviral | |
| 20 reactions | |
| TOPO Cloning Kit | |
| V5 Epitope Tag | |
| Blasticidin |
For Research Use Only. Not for use in diagnostic procedures.