The pBS-KS-attB2-SA(1)-T2A-p65AD-Hsp70 plasmid contains a recombination-mediated cassette exchange (RMCE) cassette that consists of two inverted attB sites which surround an intron phase 1 "Trojan p65AD" cassette composed of a splice acceptor site followed by the T2A peptide, sequence encoding a p65(AD)::Zip+ hemidriver (component of a 'split driver' system, contains the p65 activation domain) and an Hsp70 transcription termination signal. In the presence of phiC31:int integrase, the attB cassette can undergo RMCE with a target in the genome that contains inverted attP sites, such as the Mi{MIC} element, replacing the target attP cassette with the donor attB cassette sequence. Integration of the cassette into a coding intron (with the same phase) of a native Drosophila gene of interest will result in the cassette behaving as a "Trojan" exon: the splice acceptor site ensures that the T2A-p65(AD)::Zip+ open reading frame is incorporated into the mRNA of the native Drosophila gene, while the T2A sequence truncates the native gene product and promotes the separate translation of the p65(AD)::Zip+ open reading frame. Thus the p65(AD)::Zip+ hemidriver should be expressed under the control of the regulatory sequences of the native Drosophila gene of interest in the resulting fly line.
One of 3 essentially identical plasmids (pBS-KS-attB2-SA(0)-T2A-p65AD-Hsp70, pBS-KS-attB2-SA(1)-T2A-p65AD-Hsp70 and pBS-KS-attB2-SA(2)-T2A-p65AD-Hsp70) that vary only in the phase of the Trojan p65AD cassette.