Accurately estimating consumptive water use in the Colorado River Basin (CRB) is important for assessing and managing limited water resources in the basin. Increasing water demand from various sectors may threaten long-term sustainability of the water supply in the arid southwestern United States. We have developed a first-ever basin-wide actual evapotranspiration (ETa) map of the CRB at the Landsat scale for water use assessment at the field level. We used the operational Simplified Surface Energy Balance (SSEBop) model for estimating ETa using 328 cloud-free Landsat images acquired during 2010. Our results show that cropland had the highest ETa among all land cover classes except for water. Validation using eddy covariance measured ETa showed that the SSEBop model nicely captured the variability in annual ETa with an overall R2 of 0.78 and a mean bias error of about 10%. Comparison with water balance-based ETa showed good agreement (R2 = 0.85) at the sub-basin level. Though there was good correlation (R2 = 0.79) between Moderate Resolution Imaging Spectroradiometer (MODIS)-based ETa (1 km spatial resolution) and Landsat-based ETa (30 m spatial resolution), the spatial distribution of MODIS-based ETa was not suitable for water use assessment at the field level. In contrast, Landsat-based ETa has good potential to be used at the field level for water management. With further validation using multiple years and sites, our methodology can be applied for regular production of ETa maps of larger areas such as the conterminous United States.