Primary Identifier | MGI:6759816 | Allele Type | Targeted |
Attribute String | Conditional ready, Epitope tag, Inserted expressed sequence, Reporter | Gene | Gt(ROSA)26Sor |
Transmission | Germline | Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Is Recombinase | false | Is Wild Type | false |
molecularNote | 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 His-H2B-Cherry gene (monomeric DsRed fluorescent protein variant mCherry with N-terminal 6xHis affinity tag and 125 amino acid human histone 2B chromatin/nucleus tag sequence), 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-V5 gene (enhanced green fluorescent protein with 127 amino acid human histone 2B chromatin/nucleus tag plus C-terminal paramyxovirus V5 tag sequence), in-frame T2A sequence, in-frame sequence for a dominant-negative form of mouse Maml1 (DN-Maml1 coding for amino-acids 13-74), 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 tag and 125 amino acid human histone 2B chromatin/nucleus tag sequence), in-frame T2A sequence, in-frame mouse Notch1 intracellular domain (NICD) and PEST domain sequence (NICD-PEST), WPRE sequence, SV40 polyA signal sequence, an FRT site flanked neomycin resistance gene cassette, and a second 2xINS sequence. |