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(Search Vector Database)
Mammalian Expression, Lentiviral, Cre/Lox
MCS - see map
List of Sequencing Primers
Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
Notes from Addgene (1)
Ancillary Agreement for Plasmids Containing FP Materials
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.
Addgene has sequenced a portion of this plasmid for verification.
Full plasmid sequence is available only if provided by the depositing laboratory.
Please acknowledge the principal investigator and cite this article if you use
this plasmid in a publication. Also, please include the text "Addgene plasmid
12257" in your Materials and Methods section.
Price: US $65