The pCFD6.FRT vector is designed to facilitate cloning of one or multiple Cas9-specific guide RNAs (gRNAs) downstream of 10xUAS regulatory sequences and the Drosophila Synthetic Core Promoter (DSCP) promoter. The gRNAs are flanked by tRNAs and are initially expressed as a single polycistronic tRNA-gRNA cassette; after this primary transcript is expressed, the tRNAs should form secondary structures that are recognized by the endogenous tRNA-processing machinery (RNase enzymes P and Z), resulting in the release of the gRNAs (PMID:25733849). The vector contains an attB site, allowing integration into an attP-containing docking site in the genome to produce a transgenic construct. pCFD6.FRT also contains two non-compatible FRT sites either side of the sgRNA cassette (FRT2 at the N-terminal end and FRT5 at the C-terminal end); this allows for future in vivo exchange of sequences upstream or downstream of the sgRNAs.