Introduction: Built and natural environments may provide opportunities for physical activity. However, studies are limited by primarily using residential addresses to define exposure and self-report to measure physical activity. We quantified associations between global positioning systems (GPS)-based activity space measures of environmental exposure and accelerometer-based physical activity. Methods: Using a nationwide sample of working female adults (N = 354), we obtained seven days of GPS and accelerometry data. We created Daily Path Area activity spaces using GPS data and linked these activity spaces to spatial datasets on walkability (EPA Smart Location Database at the Census block group level) and greenness (satellite vegetation at 250 m resolution). We utilized generalized additive models to examine nonlinear associations between activity space exposures and accelerometer-derived physical activity outcomes adjusted for demographic characteristics, socioeconomic factors, and self-rated health. Results: Higher activity space walkability was associated with higher levels of moderate-vigorous physical activity, and higher activity space greenness was associated with greater numbers of steps per week. No strong relationships were observed for sedentary behavior or light physical activity. Highest levels of moderate-vigorous physical activity were observed for participants with both high walkability and high greenness in their activity spaces. Conclusion: This study contributes evidence that higher levels of physical activity occur in environments with more dense, diverse, and well-connected built environments, and with higher amounts of vegetation. These data suggest that urban planners, landscape architects, and policy makers should implement and evaluate environmental interventions to encourage higher levels of physical activity.