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pWPXL
(Plasmid #12257)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 12257 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

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

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • 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
  • (Common Sequencing Primers)

Resource Information

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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pWPXL was a gift from Didier Trono (Addgene plasmid # 12257)