A loxP site was inserted 40 nucleotides 5' of the initiation methionine codon in exon 3 and a floxed neomycin resistance gene as well as an EGFP gene were inserted in the intron between exons 5 and 6. The EGFP insert was non functional. Cre mediated recombination removed the neo cassette leaving the loxP sites in exon 3 and intron 5 intact. (J:94941)