The P{CFD4FLPOUT-Gprk1-Gprk2} 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 (Gprk1U6.Gprk1+Gprk2.CFD4FLPOUT) - FRT3 site - FRT site - sgRNA2 (Gprk2U6.Gprk1+Gprk2.CFD4FLPOUT) - Disc\RFPmCh.3xP3.cUa marker - FRT3 site.
U6 promoter sequences drive expression of a Gprk1 sgRNA (GGCTGATGTCAGCTATTTAATGG).
U6 promoter sequences drive expression of a Gprk2 sgRNA (TGACATCGGCCGTCGCTTCTTGG).