Venomous marine gastropods of genus Conus utilize one of the most sophisticated envenomation strategies known, with a single venom comprising of complex mixture of peptides evolved to target a diverse array of neuronal receptors, ion channels and transporters with high affinity and specificity. All of the ~ 800 species typically express ~ 100 dominant peptides and an additional 1000–8000 related variants arising from transcriptomic and processing messiness, with most pharmacologically uncharacterized. To better understand how cone snail have evolved separate venoms for predation and defense, we have established functional screens and a venom peptide discovery pipeline that integrates high-resolution venom transcriptomics with state-of-the-art proteomics. Using this approach, we identified key components of the motor, nirvana and lightning-strike cabals, allowing us to piece together the critical bioactive components deployed in predation and/or defense and identify novel conotoxins with potential as research tools and/or leads to new therapeutics.