The dataset gathers available regulations of human activities and protection levels of Marine Protected Areas (MPAs) of the European Union (EU). The MPA list and polygons were extracted from the MPA database of the European Environment Agency (EEA) and completed with available zoning systems (all were filtered for their marine area reported under the Marine Strategy Framework Directive). Fully-overlapping MPAs were merged. In the resulting dataset, MPA features are provided (gathered from EEA, WDPA, ProtectedSeas), including the year of designation, designation types (e.g., national, Natura 2000) and subtypes (e.g., reserves, national parks), database identifiers (WDPA, Natura 2000, OSPAR, etc.), IUCN categories, and main protection focus. We provide summarized data on maritime activities that overlap with MPA polygons from two types of datasets: activities-focused datasets (national marine spatial plans, and additional European and regional databases, like EMODnet) and MPA-focused datasets gathering data from management plans (ProtectedSeas, expert-based assessments about OSPAR and Portuguese MPAs). This dataset therefore compiles data that could be gathered from accessible legal frameworks regarding aquaculture, fisheries, anchoring, infrastructures (including harbors and renewable energy), mining, transport, coastal land-based uses (desalinization, sewage plants) and other non-extractive uses (e.g., recreational), making them readily accessible. Using the MPA Guide classification system, we computed two scenarios of potential impact for each activity, which were used to assess two scenarios of protection levels per MPA. Some MPAs could not be associated with any MPA features, regulations, or protection levels. Finally, we detail the protocol to match information from multiple databases (e.g., with MPA polygons formatted differently) and provide a quality check by comparing this dataset to previous assessments. This dataset was used to analyze MPAs’ protection levels across countries, regions and MPA features (e.g., IUCN categories, designations). It was also used to investigate the sources of information available and the levels of regulations for each maritime activity in EU MPAs. This dataset can therefore be used for further analyses on the use of EU MPAS to regulate activities and to compare with future assessments or with additional data we did not have access to (e.g., gathered at national scale). Such research is crucial to plan and monitor the implementation of the EU 2030 Biodiversity Strategy, targeting 10% of strictly protected MPAs in each sea region.