Igs2^{tm5(CAG-tTA2,-TagBFP)Luo}

A CAG-lox-STOP-lox-tTA2-T2A-H2B-TagBFP-3xMyc-WPRE insertion was targeted to the mouse Igs2 locus via phiC31 integrase-mediated transgenesis. The embryonic stem cells used were previously targeted with the H11 allele ( Igs2tm3.1Luo) or H11P3; a unique sequence from the promoter of the yeast his3 gene followed by three shortened tandem attP sites and one FRT5 site. tTA2 is an enhanced variant of tTA (tetracycline-controlled transactivator) in which the potent activation domain (V16) is replaced with three minimal activation domains of about 14 amino acids each. Viral 2A oligopeptide (T2A) mediates ribosomal skipping, prevents covalent linking, and enables continued translation. mTagBFP is a monomeric blue fluorescent protein that generates bright blue fluorescence with excitation/emission maxima at 402 and 457 nm, characterized by high photostability and extremely high pH-stability. mTagBFP was derived from TagRFP, a monomeric red (orange) fluorescent protein generated from the wildtype RFP from sea anemone Entacmaea quadricolor. (J:101977)