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They are built of an N\u2010terminal linear cationic domain (\u223c40 residues) and a C\u2010terminal knottin domain (\u223c60 residues). The linear domain empowers spiderines with strong cytolytic activity. In the present work we report 16 novel spiderine sequences from <jats:italic>Oxyopes takobius<\/jats:italic> and <jats:italic>Oxyopes lineatus<\/jats:italic> classified into two subfamilies. Strikingly, negative selection acts on both linear and knottin domains. Genes encoding <jats:italic>Oxyopes<\/jats:italic> two\u2010domain toxins were sequenced and found to be intronless. 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