UAS regulatory sequences drive expression of Esyt2-RB cDNA (BDGP RE26910) in which the conserved aspartates in each C2 domain are mutated into asparagine (D364N, D374N, D421N, D423N, E429Q for C2A; D517N, D564N for C2B; D746N, D752N for C2C) so they are unable to bind Ca[2+]; tagged with 3xTag:FLAG at the C-terminal end.