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Using a recombineering approach, a loxP site flanked neomycin resistance gene was inserted into intron 1. An FRT site flanked neo cassette and a loxP site were inserted into intron 2. The two neo cassettes were removed through subsequent cre-mediated and flp-mediated recombination, leaving a conditional-ready allele with exon 2 floxed. (J:288972)