Gt(ROSA)26Sor^{tm65.2(CAG-tdTomato)Hze}

The targeting construct contains (from 5' to 3'): a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), a frt-flanked STOP cassette (with a short open reading frame, followed by translational stops in all three reading frames, a synthetic polyA sequence, a polyA sequence from the human growth hormone gene, and a polyA sequence from the Herpes Simplex Virus TK gene), a loxP-flanked STOP cassette (with stop codons in all three reading frames linked to three repeats of the SV40 polyA signal), a sequence encoding the tdTomato fluorescent protein (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein [tandem dimer]), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal and an AttL site.This entire targeting vector was inserted between exons 1 and 2. PhiC31-mediated recombination removed the AttB/AttP-flanked PGK-Neo-polyA cassette and replace it with the recombined AttB/AttP site (AttL). This entire targeting vector was inserted between exons 1 and 2. PhiC31-mediated recombination removed the AttB/AttP-flanked PGK-Neo-polyA cassette and replaced it with the recombined AttB/AttP site (AttL). Cre-mediated recombination removed the floxed STOP. (J:260362)