The current UAS-ORF library was created with the Gateway cloning approach, followed by ΦC31 integrase-mediated insertion of the transgenes into an attP landing site on the 3rd chromosome, i.e. all the lines in this library have the identical integration site (3R, attP-86Fb). 

 

This UAS-ORF collection mainly consists of the following groups of genes:

  • Genes implicated in growth, cell size regulation and cell cycle progression. For a considerable number of these genes, we can offer a transgenic version with its native stop codon as well as a version with a C-terminal 3xHA epitope tag.
  • Around 550 transcription factors, all of them cloned with the plasmid pGW-HA.attB and thus C-terminally tagged by a 3xHA tag.
  • A collection of single-exon genes.

 

All the ORFs are provided with a Kozak consensus sequence for efficient translation. As a 3’UTR trailer, the tubulin alpha 1 sequence is used. The sequences of the expression vectors used can be found here: pUASg.attB, pUASg-HA.attB and pGW-HA.attB. A detailed description of all aspects of this UAS-ORF library was recently published (Bischof et al., Development, 2013).