Edgar

This line stably expresses a multi-cistronic RNA encoding a nuclear EGFP fused to an E2f1 degron and unlocalized mRFP1 fused to a CycB degron. The coding sequences are separated by a T2A sequence, which causes cleavage of the primary RNA into two functional mRNAs. Thus the two coding sequences are produced in equimolar amounts, translated independently, and their protein products degraded at different times in the cell cycle. G1 cells fluoresce green in the nucleus, S phase cells red throughout, and G2 cells yellow in the nucleus (from the combination of the two fluors) and red in the cytoplasm. See Zielke et al. 2014 for full details. Fluorescence intensity is adequate for imaging and for FACS analysis without additional staining.