Opportunity costs of conservation represent the foregone economic benefits from using land for nature conservation rather than alternative activities. Considering opportunity costs therefore helps in identifying cost-effective solutions in conservation prioritisation analyses. Since comprehensive, high-resolution, pan-European opportunity cost data are currently lacking, we created a 1 km European layer of opportunity costs (€/ha/yr) for arable, pastoral, forestry and urban lands. To create this layer, we estimated the opportunity costs of productive lands (agriculture and forestry) based on (sub)national land and resource rent data, which we allocated to the grid level integrating agricultural and forestry yield data with country-specific commodity prices. We estimated opportunity costs of urban land based on area-standardised residential rents and urban population density using empirical data from 42 European residential areas. Across land types, urban land comes with the highest opportunity costs, followed by arable, pastoral, and forestry land. We envisage this new opportunity cost layer to be particular useful for broad-scale European conservation prioritisation analyses.