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The current global gold rush, driven by increasing consumption in developing countries and uncertainty in financial markets, is an increasing threat for tropical ecosystems. Gold mining causes significant alteration to the environment, yet mining is often overlooked in deforestation analyses because it occupies relatively small areas. As a result, we lack a comprehensive assessment of the spatial extent of gold mining impacts on tropical forests. In this study, we provide a regional assessment of gold mining deforestation in the tropical moist forest biome of South America. Specifically, we analyzed the patterns of forest change in gold mining sites between 2001 and 2013, and evaluated the proximity of gold mining deforestation to protected areas (PAs). The forest cover maps were produced using the Land Mapper web application and images from the MODIS satellite MOD13Q1 vegetation indices 250 m product. Annual maps of forest cover were used to model the incremental change in forest in ~1600 potential gold mining sites between 2001–2006 and 2007–2013. Approximately 1680 km² of tropical moist forest was lost in these mining sites between 2001 and 2013. Deforestation was significantly higher during the 2007–2013 period, and this was associated with the increase in global demand for gold after the international financial crisis. More than 90% of the deforestation occurred in four major hotspots: Guianan moist forest ecoregion (41%), Southwest Amazon moist forest ecoregion (28%), Tapajós–Xingú moist forest ecoregion (11%), and Magdalena Valley montane forest and Magdalena–Urabá moist forest ecoregions (9%). In addition, some of the more active zones of gold mining deforestation occurred inside or within 10 km of ~32 PAs. There is an urgent need to understand the ecological and social impacts of gold mining because it is an important cause of deforestation in the most remote forests in South America, and the impacts, particularly in aquatic systems, spread well beyond the actual mining sites.