Invitrogen™ pLenti6.3/V5-DEST™ Gateway™ Vector Kit

Catalog No.	Quantity
V53306	1 kit

Catalog No. V53306 Supplier Invitrogen™ Supplier No. V53306

Description

pLenti6.3/V5-DEST™ Gateway™ Vector Kit

Invitrogen™ 's pLenti6.3⁄V5-DEST™ Gateway™ Vector Kit is part of our ViraPower™ HiPerform™ Lentiviral Gateway™ Expression Kit (catalog K5330-00).

The pLenti6.3⁄V5-DEST™ Gateway™ Vector Kit contains the Gateway™-adapted ViraPower™ HiPerform™ lentiviral expression vector, pLenti6.3⁄V5-DEST™ for easy recombination-based cloning and high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti6.3⁄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.

Advantages

Stable expression

Long-term experiments

Accurate titer of functional virus

Flexible and versatile Gateway™ recombination cloning technology

Key Features

HiPerform™ WPRE and cPPT elements

CMV promoter

V5 epitope tag at C terminus

Blasticidin selection

Kit includes

A lacZ vector as a positive control, pLenti6.3⁄V5-GW⁄lacZ

Stbl3™ competent cells

For research use only. Not intended for any therapeutic or diagnostic use.

Specifications

• Constitutive or Inducible System: Constitutive
• Delivery Type: Lentiviral
• Promoter: CMV
• Product Type: Kit, Vector
• Selection Agent (Eukaryotic): Blasticidin
• Content And Storage: Vector Kit (Box 1)
  pLenti6.3⁄V5-DEST™ (150 ng⁄μL): 6 μg, store at -20°C
  pLenti6.3⁄V5-GW⁄lacZ Control vector (500 ng⁄μL): 10 μg, store at -20°C
  
  One Shot™ Stbl3™ Chemically Competent E. coli (Box 2)
  Stbl3™ Cells: 21 x 50 μL, store at -80°C
  pUC19 control DNA (10 pg⁄μL): 50 μL, store at -80°C
  S.O.C. Medium: 6 mL, store at -80°C
• Cloning Method: Gateway
• Quantity: 1 kit
• Vector: pDEST, pLenti
• Product Line: Gateway, HiPerform, ViraPower
• For Use With (Application): Viral Expression

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.

I sequenced one of your vectors after PCR amplification and observed a difference from what is provided online (or in the manual). Should I be concerned?

Our vectors have not been completely sequenced. Your sequence data may differ when compared to what is provided. Known mutations that do not affect the function of the vector are annotated in public databases.

Are your vectors routinely sequenced?

No, our vectors are not routinely sequenced. Quality control and release criteria utilize other methods.

How was the reference sequence for your vectors created?

Sequences provided for our vectors have been compiled from information in sequence databases, published sequences, and other sources.

Do you offer pLenti6.3/V5-DEST Gateway vector as a stand-alone product?

Unfortunately, we do not offer the pLenti6.3/V5-DEST Gateway vector as a stand-alone product. The pLenti6.3/V5-DEST Gateway vector is offered as a component of the pLenti6.3/V5-DEST Gateway Vector Kit (Cat. No. V53306) or as part of the ViraPower HiPerform Lentiviral Gateway Expression Kit (Cat. No. K5330-00).

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