M{Act5C-Cas9.P.w-.RFP+} was generated in vivo from M{Act5C-Cas9.P}, by using P{white-eraser} to remove the miniwhite marker.
A 4.4 kb fragment of the upstream regulatory region of the Act5C gene drives expression of human codon optimized Cas9 endonuclease (cloned from Addgene plasmid 41815).