Gt(ROSA)26Sor^{tm35.1(CAG-aop3/GFP)Hze}

The Rosa-CAG-LSL-Arch-GFP-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), an Arch-GFP fusion gene, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. The Arch-GFP fusion gene is flanked with small domain modifications designed to facilitate correct processing and localization of newly synthesized Arch-GFP protein. Specifically, this ss-Arch-GFP-ER2 construct was designed with the signal sequence (ss) from truncated MHC class I antigen (amino acids MVPCTLLLLLAAALAPTQTRA), a mammalian codon-optimized cDNA sequence encoding the Archaerhodopsin-3 protein (Arch, aR-3, or aop3 ; derived from the halophilic bacterium Halorubrum sodomense) fused in-frame via a 15 nucleotide linker to the amino terminus of a green fluorescent protein (GFP), and an endoplasmic reticulum exporting sequence (ER2) from the potassium channel Kir2.1 gene (amino acids FCYENEV) at the C-terminal of the GFP. PhiC31-mediated recombination replaced the neo cassette with the recombined attB/attP site (attL). (J:172633, J:182204)