Cnidaria is one of the most ancient lineages of Metazoa and is characterized by the production of venom in specialized cells. The acquisition and diversification of venom plays a key role in the evolutionary success of the entire phylum. However, research on Cnidaria venom has been focused on sea anemones, belonging to the subclass Hexacorallia. Scarce information is available on the venom produced by Octocorallia, or soft corals, and it mostly pertains to their small molecules content. Polyps of soft coral colonies are equipped with nematocyst-rich tentacles used to feed on zooplankton and deter potential predators. The lack of information regarding toxins of soft corals is a bias towards the understanding of the evolutionary history of Anthozoan venom toxins. We used a proteo-transcriptomic approach to investigate the venom of Eunicella singularis, a Mediterranean soft coral inhabiting rocky bottoms at 20-30 m depth. We identified potential cytolytic and neurotoxic proteins, finding similarities with well characterized families of cnidarian toxins. In addition, we detected uncommon polypeptides with novel cysteine frameworks. Comparative genomics analyses showed that some of these toxins are widespread across soft corals or either within Anthozoa, other are restricted to specific Octocorallia clades. Structure models of the previously uncharacterized toxins were analyzed to understand their possible function. Our results provide the first description of the toxins from a soft coral’s venom, highlighting their potential activity and expanding our understanding of the evolutionary history of venom protein families in Anthozoa