In 2013, the Regional Stream Quality Assessment (RSQA) study was started as part of the U.S. Geological Survey (USGS) National Water Quality Assessment (NAWQA) project. One of the objectives of the RSQA is to characterize the relationships between water-quality stressors and stream ecology and subsequently determine the relative effects of these stressors on aquatic biota within the streams (Garrett and others, 2017; Journey and others, 2015; Coles and others, 2019; Sheibley and others, 2017; May and others, 2020). The study was implemented in five regions across the U.S.; the Midwest (MSQA) in 2013, the southeast (SESQA) in 2014, the Pacific Northwest (PNSQA) in 2015, the northeast (NESQA) in 2016, and California (CSQA) in 2017. To meet this objective, a framework of fundamental geospatial data was required to develop physical and anthropogenic characteristics for each study region. This data release provides shape files of the four fundamental geospatial data layers developed for the RSQA and includes the regional study-area boundaries, the sampled sites, and their associated watersheds and riparian zones.