The PCR check of the insertion gave the expected sized band for the left flank and a shorter than expected sized product for the right flank.

A TI{CRIMIC.TG4.2} DNA cassette has been inserted into CG43689, in a coding intron. Not predicted to express GAL4 because the reading frame of the T2A-GAL4 of the CRIMIC donor construct is out of frame with the annotated reading frame of the protein. The TI{CRIMIC.TG4.2} cassette was inserted using the CRISPR/Cas9 technique together with a donor plasmid to drive homology directed repair. The sgRNA sequence used to target the gene was: GACGATGAGGATGTATGTAAGGG. The homology arms of the donor plasmid used were designed such that there is a small gap between the 3' end of the 5' arm and the 5' end of the 3' arm, thus the insertion of the TI{CRIMIC.TG4.2} cassette is predicted to be accompanied by a deletion of 27bp of genomic sequence flanking the insertion site.