The Mi{Trojan-lexA:QFAD.un} entry in FlyBase is used to denote the transgenic construct present in an insertion when it is known that the inserted element corresponds to one of Mi{Trojan-lexA:QFAD.0}, Mi{Trojan-lexA:QFAD.1} or Mi{Trojan-lexA:QFAD.2}, but it is not known which of these 3 intron phase variants the inserted element corresponds to. The Mi{Trojan-lexA:QFAD.un} element is generated in vivo by using phiC31:int-mediated recombination to replace the attP cassette of a Mi{MIC} element insertion with a 'Trojan lexA:QFAD ' cassette. The Trojan lexA:QFAD cassette consists of a splice acceptor site followed by the T2A peptide, sequence encoding a lexA::QF driver and an Hsp70 transcription termination signal. Integration of the cassette into a Mi{MIC} insertion in 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-lexA::QF 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 lexA::QF open reading frame. Thus the lexA::QF driver should be expressed under the control of the regulatory sequences of the native Drosophila gene of interest in the resulting fly line.
This transgenic construct is generated in vivo by using phiC31-mediated recombination to replace the attP cassette of a Mi{MIC} element insertion with an unknown phase "Trojan lexA:QFAD " cassette. The Trojan lexA:QFAD cassette consists of a splice acceptor site followed by the T2A peptide, the Ecol\lexA DNA-binding domain fused to the Ncra\QF activation domain (" lexA:QFAD ") and an Hsp70 transcription termination signal. Integration of the cassette into a Mi{MIC} insertion in 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- lexA:QFAD 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 lexA:QFAD open reading frame. Thus lexA:QFAD should be expressed under the control of the regulatory sequences of the native Drosophila gene of interest in the resulting fly line.