pWPXL
(Plasmid #12257)

• Purpose: (Empty Backbone) 2nd generation lentiviral transfer plasmid
• Depositing Lab: Didier Trono
• Publication: Trono Lab Constitutive Lentiviral Plasmids (unpublished)

Backbone

• Vector backbone: pWPXL
• Backbone size (bp): 10510
• Vector type: Mammalian Expression, Lentiviral, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Growth instructions: Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: None
• Tag / Fusion Protein:
  • EGFP

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: MCS - see map (not destroyed)
• 3′ cloning site: MCS - see map (not destroyed)
• 5′ sequencing primer: See map

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest
• Articles Citing this Plasmid:
  • 80 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pWPT or pWPXL can be used for constitutive transgene expression.
pWPXL contains the EF-1alpha promoter + intron that gives you high expression, as RNA loves to be spliced (it goes more efficiently out of the nucleus). pWPT contains only the EF-1alpha promoter

The loxP site in the 3'LTR is duplicated to the 5'LTR during reverse transcription in the target cells. This allows for removal (if necessary) of an integrated provirus by Cre.

Unique restriction sites at key positions will allow you to change promoter and transgene. PmeI is a popular site for cloning. You can also use PacI and SwaI.

Please note that ClaI in this vector is blocked by Dam methylation. This plasmid needs to be grown in a Dam- bacteria strain if you wish to use ClaI for cloning.

Packaging plasmids for Trono lab lentiviral vectors are also available at Addgene http://www.addgene.org/rnaitools

Please note that the full sequence for this plasmid is approximated and not fully verified. Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more. See LentiWeb http://www.lentiweb.com for discussions on cloning strategies and protocols.

Information for Cloning Grade DNA (Catalog # 12257-DNA.cg) ( Back to top)

Item is discontinued.

Addgene no longer distributes this item. Contact for more information.

2 µg of cloning grade DNA in Tris buffer

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here