DescriptionpCAGGS-FLPe-IRESpuro is an improved vector for expression of an enhanced FLP recombinase in mouse ES cells and oocytes. FLPe shows an enhanced thermo
stability at 37°C compared to the normal FLP recombinase. After micro-injection of pCAGGS-FLPe into oocytes approximately one-third of heterozygotic mice born show complete FLP recombination.

pCAGGS-FLPe-IRESpuro carries a strong, constitutive expression cassette for an in vitro-evolved FLP recombinase under the control of the chicken-s-actin promoter and an hCMV immediate early enhancer. The use of the chimeric CMV enhancer/s-actin promoter leads to a ubiquitous expression profile in eukaryotes.

The FLPe recombinase carries a minimal, nuclear localization signal from the SV40 larger T antigen and is operatively linked by an IRES to a puromycin resistance gene.

Additional elements of the expression cassette are an intron with splice donor and acceptor sites upstream of the FLPe gene and a bovine growth hormone polyadenylation signal downstream of the puromycin resistance gene.

The pCAGGS-FLPe plasmid is appropriate for efficient excision of DNA stretches flanked by FRT sites e.g. for excision of a resistance cassette in a conditional allele in eukaryotic cells. The plasmid carries an ampicillin resistance cassette.

ContentspCAGGS-FLPe: expression plasmid for FLPe recombinase (0.2 ug/ul, 20 ul)
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