Several resources were used to identify the protein sequence features of the Dispanins. NetPhos 2.0 [31] identified potential phosphorylation sites. NetNGlyc 1.0 and NetOGlyc 3.1 [32] were used to find possible N-and O-glycosylation sites respectively. Transmembrane helix prediction was made using TMHMM 2.0 [33] . Motifs were found manually through Jalview 2.5.1 and the server MEME 4.4.0. Finally, EMBOSS:cons was used to create consensus sequences of the different Dispanin families that emerged from the phylogenetic analysis and the bacterial sequences. The consensus sequences were aligned together with the M. brevicollis and the invertebrate sequences. The resulting MSA was viewed and trimmed in Jalview and the conserved region around the TM helices was kept. Splice sites were detected in the eukaryotic Dispanins by studying their annotation in the respective databases and align them to their genome using BLAT at the UCSC Genome Browser website (http://genome.ucsc.edu).