The P{CFD4FLPOUT-Ptp99A-PTP-ER} construct contains two sgRNAs, with each flanked by a distinct fluorescent reporter. The transgene also contains two pairs of mutually incompatible FRT sites (FRT and FRT3) arranged such that a single FLP-mediated recombination event excises one of the two sgRNAs (plus one of the reporters) in a stochastic manner, with no further recombination possible. The construct has the following structure: FRT site - Avic\GFP3xP3.cUa marker - sgRNA1 (Ptp99AU6.Ptp99A+PTP-ER.CFD4FLPOUT) - FRT3 site - FRT site - sgRNA2 (PTP-ERU6.Ptp99A+PTP-ER.CFD4FLPOUT) - Disc\RFPmCh.3xP3.cUa marker - FRT3 site.
U6 promoter sequences drive expression of a Ptp99A sgRNA (TATGTGGAACACATGGTAACCGG).
U6 promoter sequences drive expression of a PTP-ER sgRNA (TCAAGTGGGGTCGTGTCGGAGGG).