Database (refseq_protein, 30 July 2018) searches were performed with PSI-BLAST (49). Representative sequences were aligned with MUSCLE (50) and displayed with JALVIEW (51). The phylogenetic relationship of species with CATACOMB homologs was estimated by TimeTree (52), with Tupaia glis representing the Tupaia genus in TimeTree. The tree was visualized at the iTOL server (53). Marsupial and Monotreme genomes were also searched with tblastn using CATACOMB sequences from various eutherian clades to exclude the possibility that the failure to identify CATACOMB homologs outside eutheria with PSI-BLAST was due to lack of proper gene prediction.

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