A map of ecological regions of the conterminous United States, first published in 1987, has been greatly refined and expanded into a hierarchical spatial framework in response to user needs, particularly by state resource management agencies. In collaboration with scientists and resource managers from numerous agencies and institutions in the United States, Mexico, and Canada, the framework has been expanded to cover North America, and the original ecoregions (now termed Level III) have been refined, subdivided, and aggregated to identify coarser as well as more detailed spatial units. The most generalized units (Level I) define 10 ecoregions in the conterminous U.S., while the finest-scale units (Level IV) identify 967 ecoregions. In this paper, we explain the logic underpinning the approach, discuss the evolution of the regional mapping process, and provide examples of how the ecoregions were distinguished at each hierarchical level. The variety of applications of the ecoregion framework illustrates its utility in resource assessment and management.