SynBlast pipeline steps. (1) A focal protein coding gene of interest and its surrounding genes (within a certain flanking size) are selected as reference set. Protein sequences and genomic positional information are either compiled manually or retrieved from Ensembl using the tool getEnsemblProteins.pl. (2) tblastn searches of all reference proteins are performed against selected target genome databases. (3) The resulting blast hit tables are scanned for regions of possibly conserved gene content. These regions are stored in separate blast hit tables. (4) The resulting sets of possibly syntenically conserved blast hits are evaluated based on their sum of blast bit-scores obtained by means of a gene loci order alignment to the reference set. The final decision on orthology or paralogy of ranked syntenic blocks is left to the user.