A set of stocks in which the coding sequence of a gene of interest has been replaced by a KozakGAL4 promoter trap sequence, resulting in a simultaneous knock-out of the gene of interest and a knock-in of GAL4 driver sequence that is expected to be expressed under the control of the endogenous regulatory sequences of the targeted gene. The cassette was inserted via the technique described in Kanca et al., 2022, eLife 11: e76077 (FBrf0253899).
The target gene is Rab9D. The 200bp 5' HA aligns 100% to X:10589171 in Rab9D, but the 3' end (positions 126-200) of the 5' HA aligns 100% to 3 other sites at corresponding positions in the 5' UTR of Rab9Db, Rab9E, Rab9Fa. The 200bp 3' HA aligns 100% to the 3' UTR of target Rab9D plus 100% to 3' UTR of Rab9Db and Rab9Fa, and 197/200 to 3' UTR of Rab9E. The sgRNA gives a 23/23 alignment to a site in the 5' UTR of Rab9D, Rab9Db, Rab9E, Rab9Fa and Rab9Fb. The sgRNA2 gives a 23/23 alignment to site in 3' UTR of Rab9D, Rab9Db, Rab9E, Rab9Fa.
