Longstanding research suggests a positive relation between physical activity and health. However, when investigating this relation most studies focused on the absence of disease or infirmity as health indicators. The relation between physical activity and positive health-related constructs such as subjective well-being (SWB) remains oftentimes unexplored. The present meta-analysis offers a rigorous test of the relation between physical activity and SWB in healthy individuals, by including all different kinds of physical activity and SWB facets from childhood to old age. Random-effects meta-analysis using robust variance estimation revealed a positive relation (d = 0.360, 95% CI [0.301, 0.420]). Our results demonstrate a small beneficial main effect of physical activity on SWB, independent of the prior fitness level of the participants and various characteristics of the physical activity intervention. This effect was found in experimental studies as well as in correlational and quasi-experimental studies. Physical activity was more strongly related to positive affect compared to cognitive well-being and was unrelated to negative affect. Our results provide evidence for the importance of physical activity in the context of well-being. Further, we also systematically review and discuss the large heterogeneity of studies published on this relation and warrant further research regarding underlying mechanisms.