Gt(ROSA)26Sor^{tm4(CAG-mNeonGreen,-sfGFP)Xwy}

The final vector, generated through site specific integrase-mediated transgenesis, contains from 5' to 3' attL,a CAG promoter, a loxP-flanked STOP cassette, a Kozak consensus start site (for strong initiation of translation), two Myc-tags, an optimized C22-repeat MORF switch, and a farnesylated mNeonGreen fluorescent reporter, a tdTomato linker domain sequence, a superfolder GFP fluorescent reporter with several V5 epitopes (smFP-V5), polyA, and attR-attP3-FRT-R1/2 inserted in to the Gt(ROSA)26Sor locus. mNeonGreen is one of the brightest green/yellow fluorescent proteins and is derived from Branchiostoma lanceolatum. The farnesylation sequence (CAAX domain from Ras) targets the reporter to membranous structures including neuronal dendrites and axons. sfGFP was altered to embed several V5 epitopes (V5 is a 14 amino acid simian virus 5 epitope) in the GFP scaffold - 3 epitopes at the N terminus, 4 epitopes into the internal 172-173 loop and 3 epitopes at the C terminus. The inclusion of the V5 epitopes creates a highly antigenic "spaghetti monster" fluorescent protein (smGFP) that contains numerous copies of peptide epitopes and simultaneously binds IgG antibodies at each location. (J:292704)