Plasmid 12258: pWPXLd

• Gene/insert name: None
• Fusion protein or tag: EGFP
• Vector backbone: pWPXLd
  (Search Vector Database)
• Vector type: Mammalian Expression, Lentiviral
• Backbone size w/o insert: 10455
• Cloning site 5': MCS - see map
• Site destroyed during cloning: No
• Cloning site 3': MCS - see map
• Site destroyed during cloning: No
• 5' sequencing primer: See map
• Bacterial resistance(s) Ampicillin
• Growth strain(s) Stbl3
• Growth temperature (℃): 37
• Growth instructions: Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
• High or low copy: High Copy
• Sequence: View sequences (2)
• Map: View map
• Supplemental document: Notes from Addgene (1)
• Principal Investigator: Didier Trono
• Terms and Licenses: MTA

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. 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.