Gt(ROSA)26Sor^{tm2(CAG-NuTRAP)Evdr}

The targeting vector was designed with (from 5' to 3') with a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, a FRT site, a loxP-flanked STOP cassette, the E. Coli biotin ligase gene (birA), a 2A peptide, the BLRP-mCherry-mRANGAP1 fusion gene (described below), a 2A peptide, the EGFP-L10a fusion protein (an enhanced green fluorescent protein gene [EGFP] fused in-frame to the N-terminus of a cDNA sequence encoding the mouse 60S ribosomal subunit L10a [Rpl10a]), a WPRE, a polyA signal, an AttB site, a PGK-FRT-Neo-PGK polyA, and an AttP site. This entire targeting vector was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The BLRP-mCherry-mRANGAP1 fusion gene consists of a monomeric red fluorescent protein sequence fused in-frame to the N-terminal of a cDNA sequence encoding the mouse nuclear membrane RAN GTPase activating protein 1. This fusion gene is tagged at the mCherry N-terminal with a biotin ligase recognition peptide (BLRP ; a 23 amino acid protein with AviTag core. (J:101977)