The club was founded in 1965. In 2001 they joined the Senior Division of the Reading League, going on to win the league in 2006–07. As a result, the club were promoted to Division One East of the Hellenic League. After finishing fourth in their first season, they claimed the runners-up spot in 2008–09 season, earning promotion to the Premier Division. The club made its debut in the FA Vase in the 2010–11 season and the 2011–12 season saw them play in the FA Cup for the first time. Their first match was against Wembley, which was the first-ever FA Cup match streamed live on Facebook. The game also led to Ascot's then-record attendance of 1,149. In 2013–14 the club won the Hellenic League Challenge Cup, defeating Ardley United 2–1 in the final.
The 2018–19 season saw Ascot win the Challenge Cup for a second time with a 3–1 win against Windsor in the final. In 2022–23 they were Combined Counties Football League Premier Division North champions, earning promotion with 102 points as well as reaching the final of the 2022–23 FA Vase at Wembley Stadium.