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Life Technologies' Flp-In™ Complete System allows integration and expression of your gene of interest in mammalian cells at a specific genomic location. The Flp-In System involves introduction of a Flp Recombination Target (FRT) site into the genome of the mammalian cell line of choice. An expression vector containing your gene of interest is then integrated into the genome via Flp recombinase-mediated DNA recombination at the FRT site.
The major components of the Flp-In™ System include:
• A Flp-In. target site vector, pFRT⁄lacZeo, for generation of a host cell line containing an integrated FRT site • An expression plasmid containing a FRT site linked to the hygromycin resistance gene for Flp recombinase-mediated integration and selection of a stable cell line expressing your gene of interest under the control of the human cytomegalovirus (CMV) immediate-early enhancer⁄promoter • A Flp recombinase expression plasmid, pOG44, for expression of the Flp recombinase under the control of the human CMV promoter • A control expression plasmid containing the chloramphenicol acetyl transferase (CAT) gene, which when cotransfected with pOG44 into your Flp-In. host cell line, expresses CAT
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