Gt(ROSA)26Sor^{tm2(CAG-EYFP*,-mCherry*,-EGFP*,-mCerulean*)Ben}

This allele has three chromatin/nuclear-targeted fluorescent protein sequences uniquely flanked with pairs of incompatible lox sites (in the same direction), all inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The targeting vector is designed with (from 5' to 3') two copies of chicken beta-globin HS4 insulator element (2xINS; increase downstream gene expression and minimize regulatory interference), a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), a loxN site, a lox2272 site, a loxP site, the N-PhiMut gene (non-cytotoxic/non-fluorescent Phialidium yellow fluorescent protein variant PhiYFP with nuclear localization signal), SV40 polyA signal sequence, a second loxN site, a full consensus KOZAK sequence, the 6xHis-H2B-mCherry gene (monomeric DsRed fluorescent protein variant mCherry with N-terminal 6xHis affinity tag and 125 amino acid human histone 2B chromatin/nucleus tag), in-frame Thosea asigna virus 2A cleavage site (T2A; mediates ribosomal skipping), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance mRNA transcript stability), SV40 polyA signal sequence, a second lox2272 site, a full consensus KOZAK sequence, the H2B-EGFP gene (enhanced green fluorescent protein with 127 amino acid human histone 2B chromatin/nucleus tag and KOZAK sequence plus C-terminal paramyxovirus V5 tag sequence), in-frame T2A sequence, WPRE sequence, SV40 polyA signal sequence, a second loxP site, a full consensus KOZAK sequence, the HA-H2B-mCerulean gene (monomeric enhanced cyan fluorescent protein variant mCerulean with N-terminal hemagglutinin (HA) tag, 125 amino acid human histone 2B chromatin/nucleus tag and KOZAK sequence), in-frame T2A sequence, WPRE sequence, SV40 polyA signal sequence, an FRT site flanked neomycin resistance gene cassette, and a second 2xINS sequence. (J:252386)