Conservation planners use a variety of decision-making tools, many of which require identifying and prioritizing spatial units based on their biodiversity and levels of imperilment. Turtles are highly imperiled, but present schemes for determining global priority areas are focused mostly on broad regional scales. We conduct the first global evaluation of turtle biodiversity and imperilment at a sub-basin level to identify geographically smaller areas of high conservation value, and compare with these existing prioritizations. We employed two spatial analyses—bivariate maps and local indicator of spatial association (LISA)—to identify and prioritize sub-basin clusters based on multiple biodiversity and conservation metrics in addition to species richness. Most high-priority sub-basin clusters were located along tropical and subtropical coastlines. A new area of global significance for turtle conservation was identified in southwest India. Many sub-basins of the Indomalayan Realm were clustered as high or intermediate priority, with large clusters of high-priority sub-basins also in tropical Australasia. Other high and intermediate priority sub-basin clusters were found in the Afrotropical, Neotropical, and Nearctic realms, often in previously recognized turtle biodiversity hotspots. Many conservation-priority sub-basins with high turtle-species richness and endemism are in lowland and coastal areas where endemics (some from ancient lineages) are imperiled in association with a high human footprint. Our findings reiterate the global significance of Asia as a key area of chelonian conservation need, while identifying focal areas across the globe where the need for targeted turtle conservation is especially great.