Rasgrf2^{tm2.1(folA/flpo)Hze}

The targeting vector contained, from 5' to 3': a partial Rasgrf2 sequence spanning intron 29-33 (with an FRT3 site in intron 33) and partial exon 34 sequence up to (but not including) the endogenous stop codon, a viral 2A oligopeptide sequence (T2A), an in-frame dgFlpO fusion gene (described below), a bovine growth hormone polyA sequence, an AttB site, a PGK-Neomycin cassette, an FRT5 site, an mRNA splice acceptor sequence, the 3' portion of the hygromycin gene, and an AttP site. The dgFlpO fusion gene is an enhanced green fluorescent protein/FlpO recombinase fusion gene with an N terminal fusion of the first 159 amino acids of the Escherichia coli K-12 strain chromosomal dihydrofolate reductase gene (DHFR or folA) harboring the G67S mutation and modified to also include the R12Y/Y100I destabilizing domain mutations. PhiC31-mediated recombination removed the AttB/AttP-flanked sequence and replaced it with the recombined AttB/AttP site (AttL). (J:101977)