Fig. 1. Strategy for rapid systematic localization and functional characterization of proteins encoded by novel cDNAs. Individual full coding cDNAs were PCR amplified using automatically designed primers, which also added the recombination sequences attB1 and attB2 at the 5′ and 3′ ends, respectively. These products were then recombined into the entry clones, which then served as a universal source of material for all expression vectors. N- and C-terminal GFP (CFP/YFP) fusion expression vectors were both generated from the entry clone in a single recombination reaction. These clones were then transfected into cells and the localizations of the fusion proteins recorded. This information was then combined with the bioinformatic data generated from sequence analysis and additional immunostainings using compartment-specific antibodies when appropriate.