Constructing pBID-UASC, a Versatile Reporter Vector

pBID-UASC was constructed by three-fragment ligation. To this end, 255-bp 5xUAS sequence was PCR amplified from pUAST [2] (link) with Pry1 and SacI-UAS reverse primers and cloned into pCR8GW-TOPO (Invitrogen) to create pCR8GW-5xUAS. Meanwhile, a 344-bp DSCP (Drosophila Synthetic Core Promoter) sequence was PCR amplified from pBPGUw (gift from Gerald Rubin, Janelia Farm) with Gateway attR2 and EcoRI-DSCP reverse primers, and a 151-bp fragment released from the 344-bp DSCP fragment with SacI/EcoRI was cloned into SacI/EcoRI sites of pBluescript SK(+), resulting in pBS-DSCP. A 132-bp HindIII/SacI 5xUAS fragment of pCR8GW-5xUAS and a 159-bp SacI/EcoRI DSCP fragment of pBS-DSCP were inserted in HindIII/EcoRI sites of pBID-UAS, giving rise to pBID-UASC.