|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.
Vector backbonepBluescript KS+
- Backbone size w/o insert (bp) 2969
Vector typeCre/Lox ; BAC recombineering
Selectable markersNeomycin (select with G418) ; ganciclovir
Growth in Bacteria
Growth instructions1. EL350 – for recombineering transformation steps 2. DH5alfa, XL1-blue, or other competent bacterial cell line – for non-recombineering transformation steps
Copy numberHigh Copy
Insert Size (bp)3697
MutationThis 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 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)
Please note that the plasmid names and lox sites have been corrected since publication.
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