TI{CRIMIC.GT14} represents a DNA segment that has been inserted into the genome by homology directed repair using CRISPR/Cas9 in combination with a donor plasmid based on pM14. The inserted TI{CRIMIC.GT14} DNA is not flanked by transposable element ends and consists of a recombination-mediated cassette exchange (RMCE) cassette flanked by a pair of inverted attP sites. Inside the attP sites is a cassette flanked by a pair of direct FRT sites. This in turn contains a mutagenic gene trap element consisting of a splice acceptor site, stop codons in all three reading frames and an SV40 polyadenylation signal. A Avic\GFP3xP3.cLa marker allele followed by a polyadenylation signal is present downstream of the gene trap element. If the TI{CRIMIC.GT14} sequence is inserted into an intron of a native Drosophila gene of interest, the gene may be disrupted by the gene trap element. The attP flanked RMCE cassette can be replaced with DNA from a compatible donor plasmid (where the sequence to be inserted is flanked by inverted attB sites) through recombination-mediated cassette exchange (RMCE) driven by the phiC31:int integrase.
