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pWPI
(Plasmid #12254)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 12254 Standard format: Plasmid sent in bacteria as agar stab 1 $75
Cloning Grade DNA 12254-DNA.cg 2 µg of cloning grade DNA in Tris buffer 1 $95

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pWPXL
  • Backbone size (bp) 11101
  • 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
    • EMCV IRES-EGFP cassette (C terminal on backbone)

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

This bicistronic vector allows for simultaneous expression of a transgene and EGFP marker to facilitate tracking of transduced cells. The EGFP marker cDNA has been inserted downstream of EMCV IRES.

A popular cloning strategy is to clone into the PmeI site upstream of the EMCV IRES. You can also clone into the PacI or SwaI site.

If you want to put another fluorescent protein after the IRES, note that you need to make sure that the open reading frame of the second gene is precisely fused to 11th ATG of IRES (i.e., 11th ATG of IRES is a start of your cDNA).

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.

Updated sequence provided by Chang-Xin Shi, Mayo Clinic

Information for Cloning Grade DNA (Catalog # 12254-DNA.cg) ( Back to top )

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 $95 USD
  • Storage DNA can be stored at 4℃ (short term) or -20℃ (long term).

Resource Information

Quality Control

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

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:

    pWPI was a gift from Didier Trono (Addgene plasmid # 12254 ; http://n2t.net/addgene:12254 ; RRID:Addgene_12254)