pWPXLd
(Plasmid #12258)

• Purpose: (Empty Backbone)
• Depositing Lab: Didier Trono
• Publication: Trono Lab Constitutive Lentiviral Plasmids (unpublished)

Backbone

• Vector backbone: pWPXLd
• Backbone size (bp): 10455
• Vector type: Mammalian Expression, Lentiviral

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:
  • Plasmid Sequence
• Articles Citing this Plasmid:
  • 60 References

Terms and Licenses

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

Depositor Comments

pWPXLd is essentially pWPXL without the LoxP site in the 3' LTR.

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

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 these vectors is blocked by Dam methylation.
The plasmids need 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. More accurate sequence can be found in the supplement documents ("Notes from Addgene") above. 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.