P{Act5C-eSpCas9.w-.RFP+} was generated in vivo from P{Act5C-eSpCas9}, using P{white-eraser} to exchange the miniwhite marker for a Disc\RFPDsRed.3xP3.cUa marker.
Act5C regulatory sequences drive expression of the eSpCas9(1.1) RNA-guided nuclease.