PBac{ScarlessHD-DsRed} consists of a Disc\RFP^{DsRed(T1).3xP3.cHa} marker flanked by termini from the Tni\piggyBac transposable element. It is typically inserted into the genome as part of a homology-directed repair event using a donor template based on the pScarlessHD-DsRed plasmid ( Addgene:64703 ). PBac{ScarlessHD-DsRed} is then often precisely excised using piggyBac transposase, leaving only a TTAA sequence at the site of the original insertion, plus any additional mutations introduced in the original homology-directed repair event.