Igs7^{tm82.1(tetO-EGFP)Hze}

The targeting vector was designed with (from 5' to 3') an FRT3 site, two tandem copies of the chicken beta-globin HS4 insulator element (to reduce reporter gene expression in absence of transactivator protein), a modified Tet response element (TRE or tetO), a loxP-flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), the enhanced green fluorescent protein (EGFP) coding sequence, a WPRE sequence (to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, two tandem copies of the chicken beta-globin HS4 insulator element, an AttB site, a PGK 5' hygro cassette, an RNA spice donor and a FRT5 site. Embryonic stem cells, previously targeted with FRT3-AttB-PGK-neoR-polyA::FRT5::splice acceptor::3' hygro cassette::SV40 polyA:AttP in the same locus, were re-transfected with the targeting vector and Flp recombinase vector. Mutant mice were bred with Gt(ROSA)26Sortm3(phiC31*)Sor mice to remove to remove the AttB/AttP-flanked PGK-hygro-SV40pA cassette and replace it with the recombined AttB/AttP site (AttL). The Ai82D allele is therefore TIGRE-frt3-Ins-TRE-LSL-EGFP-WPRE-bGHpA-Ins-AttL. (J:101977, J:219930)