Renewable fuel standards for biofuels have been written into policy in the U.S. to reduce the greenhouse gas (GHG) intensity of transportation energy supply. Biofuel feedstocks sourced from within a regional market have the potential to also address sustainability goals. The U.S. Mid-Atlantic region could meet the advanced fuel designation specified in the Renewable Fuel Standard (RFS2), which requires a 50% reduction in GHG emissions relative to a gasoline baseline fuel, through ethanol produced from winter barley (Hordeum vulgare L.). We estimate technology configurations and winter barley grown on available winter fallow agricultural land in six Mid-Atlantic states. Using spatially weighted stochastic GHG emission estimates for winter barley supply from 374 counties and biorefinery data from a commercial dry-grind facility design with multiple co-products, we conclude that winter barley would meet RFS2 goals even with the U.S. EPA’s indirect land use change estimates. Using a conservative threshold for soil GHG emissions sourced from barley produced on winter fallow lands in the U.S. MidAtlantic, a biorefinery located near densely populated metropolitan areas in the Eastern U.S. seaboard could economically meet the requirements of an advanced biofuel with the co-production of CO2 for the soft drink industry.