The P{CFD4FLPOUT-Src42A-Src64B} 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 (Src42AU6.Src42A+Src64B.CFD4FLPOUT) - FRT3 site - FRT site - sgRNA2 (Src64BU6.Src42A+Src64B.CFD4FLPOUT) - Disc\RFPmCh.3xP3.cUa marker - FRT3 site.
U6 promoter sequences drive expression of a Src42A sgRNA (TGTCCGGCGTGTGAGGGCATGGG).
U6 promoter sequences drive expression of a Src64B sgRNA (TTCAACGGACCCCGATGATTGGG).