See additional related dataset FlyORF-3, an epitope-tagged UAS-ORF collection.
A collection of stocks of UAS-ORF transgenic recombinants that carry the ORF of interest preceded by Scer\UAS sequences, Hsp70 promoter sequences and a Kozak consensus sequence for efficient translation. All constructs also carry the w+mC mini-white marker, and all have been introduced by recombination into a site at 86F on the third chromosome. Controlled expression of the ORFs can be effected by introduction of Scer\GAL4 drivers. See also the related dataset, an epitope-tagged UAS-ORF collection, FlyORF-2.
Full-length ORFs were cloned from cDNAs from the BDGP Drosophila Gene Collections (releases 1-3) or from the SD_cDNA library (Berkeley Drosophila Genome Project). ORFs were cloned by PCR with gene-specific primers then introduced into Gateway-based vectors that contain randomized ‘barcode’ sequences. The barcoded ORF cassettes were transferred to pUASg.attB by a Gateway recombination reaction: the resulting vectors carry the ORF of interest preceded by Scer\UAS sequences, plus a Sliv\attB recombination site. Pools of ORF clones were injected into embryos of a strain carrying M{3xP3-RFP.attP}ZH-86Fb (a phiC31\attP recombination target site on the third chromosome), and a source of integrase. Surviving males were mated, then subjected to single-fly PCR amplifying the plasmid region containing the barcode, followed by sequencing. Each new line determined to carry a transgenic recombinant was balanced; repeatedly occurring lines were discarded.
