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Invitrogen™

pLenti7.3/V5-DEST™ Gateway™ Vector Kit

The pLenti7.3/V5-DEST™ Gateway™ Vector Kit contains the Gateway™-adapted ViraPower™ HiPerform™ lentiviral expression vector, pLenti7.3/V5-DEST™ for easy recombination-based cloning and high-levelRead more
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Catalog NumberQuantity
V53406
also known as V534-06
1 Kit
Catalog number V53406
also known as V534-06
Price (EUR)
3 980,00
1 kit
Quantity:
1 Kit
Price (EUR)
3 980,00
1 kit

The pLenti7.3/V5-DEST™ Gateway™ Vector Kit contains the Gateway™-adapted ViraPower™ HiPerform™ lentiviral expression vector, pLenti7.3/V5-DEST™ for easy recombination-based cloning and high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti7.3/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. In addition, the vector allows for an accurate determination of titer of functional lentivirus in just two days using Emerald Green Fluorescent Protein (EmGFP).

Advantages

  • Transient expression
  • Short-term experiments
  • High-throughput screening
  • 2 day titer of functional virus using EmGFP
  • Flexible and versatile Gateway™ recombination cloning technology

Key Features

  • WPRE (Woodchuck Posttranscriptional Regulatory Element) from the woodchuck hepatitis virus, allows increased transgene expression and cPPT (Polypurine Tract) from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome
  • Human cytomegalovirus (CMV) immediate early promoter to control high-level expression of the gene of interest
  • SV40 promoter driving expression of EmGFP
  • Emerald Green Fluorescent Protein (EmGFP) allows easy determination of Lentiviral titers by flow cytometry and monitor transduced cells.

Kit Includes

  • pLenti7.3/V5-DEST™ Gateway™ Vector box
  • One Shot™ Stbl3™ chemically competent E. coli (Cat. No. C7373-03)

Related SKUs

  • 293FT Cell Line (Cat. No. R70007)
  • ViraPower™ Lentiviral Support Kit (K4970-00)
  • ViraPower™ Lentiviral Packaging Mix (K4975-00)
  • Lipofectamine™ 2000 (11668-019, 11668-027)
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Constitutive or Inducible SystemConstitutive
Delivery TypeLentiviral
For Use With (Application)Reporter Assays, Viral Expression
Product TypeLentiviral Expression Vector
Quantity1 Kit
Reporter GeneGFP (EmGFP)
Selection Agent (Eukaryotic)None
VectorpLenti
Cloning MethodGateway
Product LineViraPower, Gateway, HIPERFORM
PromoterCMV
Protein TagV5 Epitope Tag
Unit Size1 kit
Contents & Storage
pLenti7.3/V5-DEST™ Gateway™ Vector Kit
• pLenti7.3/V5-DEST™ Gateway™ Vector (150 ng/μL), 40 μL
• pLenti7.3/V5-GW/lacZ Control Vector (0.5 μg/μL), 20 μL
Store at -20°C.

One Shot™ Stbl3™ Chemically Competent E. coli box
Transformation reagents including:
• One Shot™ Competent Cells, 20 x 50 μL single-use aliquots
• S.O.C. medium
• pUC19 supercoiled plasmid control
Store at -80°C.

Frequently asked questions (FAQs)

Can I perform the single-step protocol for the BP/LR Clonase reaction using BP Clonase enzyme and LR Clonase enzyme instead of BP Clonase II enzyme and LR Clonase II enzyme?

In the single-step protocol for the BP/LR Clonase reaction, we would not recommend substituting the BP Clonase II/LR Clonase II enzymes with BP Clonase /LR Clonase enzymes as this would result in very low recombination efficiency.

Do you have a recommended single-step protocol for BP/LR recombination?

Yes, we have come up with a single-step protocol for BP/LR Clonase reaction (http://www.thermofisher.com/us/en/home/life-science/cloning/gateway-cloning.html#1), where DNA fragments can be cloned into Destination vectors in a single step reaction, allowing you to save time and money.

How can I move my gene of interest from a Gateway-adapted expression clone to a new Destination vector as I have lost the entry clone?

We would recommend performing a BP reaction with a Donor vector in order to obtain an entry clone. This entry clone can then be used in an LR reaction with the Destination vector to obtain the new expression clone.

Can I purchase the 5X LR Clonase buffer or 5X BP Clonase buffer separately?

We do not offer the 5X LR Clonase buffer and 5X BP Clonase buffer as standalone products. They are available as part of the enzyme kits.

Do you offer Gateway vectors for expression in plants?

We do not offer any Gateway vectors for expression in plants.