The Seminole Gulf Railway is a short line freight and passenger excursion railroad headquartered in Fort Myers, Florida, that operates two former CSX Transportation railroad lines in Southwest Florida. The company's Fort Myers Division, which was previously the southernmost segment of CSX's Fort Myers Subdivision, runs from Arcadia south to North Naples via Punta Gorda, Fort Myers, Estero, and Bonita Springs. The company's other line, the Sarasota Division, runs from Oneco south through Sarasota. Seminole Gulf acquired the lines in November 1987 and operates its own equipment. The company's first train departed Fort Myers on November 14, 1987.

Southwest Florida is the region along the southwest Gulf coast of the U.S. state of Florida. The area is known for its beaches, subtropical landscape, and winter resort economy.