Eukaryotic FLPo Recombinase Expression Plasmid: pCAGGS-FLPo (Academic price, requires limited use license)

Eukaryotic FLP recombinase expression plasmid; the codon-usage of FLP recombinase was altered to improve expression in mammalian cells (Academia).The use of this product is governed by the terms and conditions of the limited use license (FLPo Material). Industrial clients require a license from Fred Hutchinson Cancer Research Center, Seattle, Washington.FLP recombinase was originally isolated from yeast and therefore shows a significantly reduced activity at 37C due to thermal instability of the protein (Buchholz F. et al., 1996). A screen for thermo-stable mutants resulted in the identification of an enhanced FLP version (FLPe), which exhibits a 4-10 fold higher activity at 37C (Buchholz F., Angrand P.O. and Stewart A.F. 1998). FLPo is a codon-optimized FLP version first described by Christopher Raymond and Philippe Soriano (2007). Its amino acid sequence is identical to that of FLPe but the codon usage was altered to improve expression in mammalian cells. It appears to be at least 10 fold more efficient than FLPe (Kranz A. et al., 2010).Our pCAGGS expression vector carries FLPo under the control of the chicken-beta-actin promoter and an hCMV immediate early enhancer. The use of the chimeric CMV enhancer/beta-actin promoter leads to a ubiquitous expression profile in eukaryotes. The addition of a Sv40 Large T nuclear localization sequence (nls) further improves the performance in mammalian cells (Schaft J. et al., 2001). The recombinases are linked to a puromycin resistance gene by an internal ribosomal entry site (IRES).The pCAGGS-FLPo plasmid allows efficient excision of DNA stretches flanked by FRT sites. The plasmid carries a puromycin resistance gene for selection in eukaryotic cells and an ampicillin resistance cassette for selection in E. coli.Contents:Recombinase expression plasmid pCAGGS-FLPo (0.2 ug/ul, 20 ul)Manual