Sequence encoding the cno intron 1 splice acceptor, the 5'UTR and start codon, full-length wild-type cno coding sequence tagged at the C-terminal end with GFP, the 3'UTR and poly(A) signal has been integrated into the attP site present in TI{TI}cnoΔRA. This results in wild-type, GFP-tagged cno being expressed under the control of endogenous cno regulatory sequences. A single loxP site and a wGMR.PHb marker have also been inserted (downstream of the cno 3'UTR) during the re-integration event. In addition, sequence from the progenitor TI{TI}cnoΔRA chromosome is still present downstream of the newly inserted sequence. This includes 1. a loxP cassette containing a Disc\RFPDsRed(T1).3xP3.cHa marker and 2. a 3' portion of cno from exon 7 onwards, encoding a truncated cno 'ΔRA' protein that lacks Ras-association (RA) domain 1 and most of RA2. This truncated 'ΔRA' protein is also expressed, in addition to the wild-type GFP-tagged protein.
