Fig. 1.

Construct designs and the principle of conversion. a) Diagrams of Flp and LexA HACK donor constructs. The vectors were constructed in pAC, a dual-transformation backbone (via PhiC31 and P-transposase) that carries a mini-white selection marker. See text for descriptions of other components. pA, polyA tail; TS, gRNA target sequence; P, P-element. b) Diagram of Gal4-to-Flp/LexA conversion using a HACK donor line. The donor expresses two gRNAs (TS1 and TS2) targeting the tissue-specific (ts) Gal4, which results in in-frame incorporation of 2A-Flp/LexA into the Gal4 locus through HDR. The donor expresses ubi-nBFP that can be selected against when screening for convertants.