brp regulatory and coding sequences (genomic fragment of ~50kb) in which the stop codon has been replaced by a single FRT site, a GFP tag, the 2A peptide and a lexA::VP16 fusion. The brp protein produced is constitutively tagged with GFP, and the presence of the 2A peptide allows the lexA::VP16 fusion to be cotranslated as a separate protein, under the control of the brp regulatory sequences.
This allele is part of the STaR system.
The PBac{brp(FRT.Stop)GFP} construct used in FBrf0223931 was originally constructed to have the structure
brp—FRT stop cassette—GFP—2A peptide— LexA::VP16
but the transformants were found to have a mutation in LexA rendering the protein nonfunctional; consequently, the 2A peptide— LexA::VP16 portion of the construct was not mentioned in the paper. The PBac{brp(-FRT)GFP} derivative obtained upon FLP-mediated recombination also carries sequences encoding the nonfunctional LexA::VP16 fusion protein.
PBac{brp-FRT-GFP-2A-LexA-VP16} was constructed as a control to show the results that would be obtained upon removal of the FRT cassette from PBac{brp(FRT.Stop)GFP}. The LexA::VP16 sequence was found to be unmutated upon transformation, so functional protein was produced.