This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

pLCA.66/2272
(Plasmid #22733)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 22733 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBluescript KS+
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 2969
  • Vector type
    Cre/Lox ; BAC recombineering
  • Selectable markers
    Neomycin (select with G418) ; ganciclovir

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    1. EL350 – for recombineering transformation steps 2. DH5alfa, XL1-blue, or other competent bacterial cell line – for non-recombineering transformation steps
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Lox66-pU(delta)TK-EM7Neo-Lox2272
  • Insert Size (bp)
    3697
  • Mutation
    This vector was designed for assembling gene targeting constructs using the BAC recombineering approach. The vector contains a puromycin-(delta)thymidine kinase fusion gene driven by the mouse phosphoglycerol kinase promoter (pUDTK) and a neomycin resistant gene driven by the bacterial EM7 promoter (EM7neo). These selectable markers are flanked by lox66 and lox2272 sites (Cre-recombinase recognition sequences) in a pBluescript KS+ backbone with a modified multi-cloning region

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Kpn1 (not destroyed)
  • 3′ cloning site Kpn1 (not destroyed)
  • 5′ sequencing primer T3
  • 3′ sequencing primer T7
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Please note that the plasmid names and lox sites have been corrected since publication.

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:

    pLCA.66/2272 was a gift from Mark Magnuson (Addgene plasmid # 22733 ; http://n2t.net/addgene:22733 ; RRID:Addgene_22733)
  • For your References section:

    Quantification of factors influencing fluorescent protein expression using RMCE to generate an allelic series in the ROSA26 locus in mice. Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R, Yuan W, Piston DW, Magnuson MA. Dis Model Mech. 2011 Feb 14. ():. 10.1242/dmm.006569 PubMed 21324933