Size distribution of the 5570 predicted P. aeruginosa PA01 ORFs (A). Schematic representation of the two-stage PCR reaction strategy used for amplification of the P. aeruginosa ORFs (B). Amplification of an ORF (purple) during the first PCR reaction and generation of the attB recombination sequences (green) during the first and second PCR reactions by use of overlapping primers are indicated. Schematic representation of the configuration of the amplified ORFs in the repository (C) showing attB1 and attB2 Gateway recombination sites, Ribosomal binding site (RBS), Kozak sequence. Normalized initiation codons (ATG) are indicated at the 5′ end of the ORF, and stop-Gly substitution at the 3′ end of the ORFs is also indicated.
