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Maeson Vaughan, a former Year 11 student of the school was named in the senior matchday squad for Oxford United against Stevenage.
He came off the bench in the 90th minute, becoming the youngest player in the club’s history to feature in an FA Cup match.
He played nine minutes in the first round fixture at just 16 years old.
Adam Bayliss, head of PE at Theale Green School, said: "Maeson, everyone at Theale Green School is incredibly proud of you for being selected to represent Oxford United Football Club and making your first-team senior debut against Stevenage.
"This is just the beginning, and we can't wait to watch you pull on an Oxford shirt many more times in the future.
"Keep working hard, keep believing in yourself and most importantly, enjoy every minute of it."
Maeson’s debut is the latest milestone in a promising football journey, having impressed throughout Oxford United’s academy.
He was one of several academy graduates named in the squad, with 14 of the 20 players on the day having come through the club’s youth system.
Theale Green School described Maeson’s achievement as a testament to his 'exceptional talent and work ethic,' and said it is an inspiration to current students.
A spokesperson for the school said: "Maeson’s achievement is an inspiration to current students and demonstrates what can be accomplished through determination and a commitment to pursuing your ambitions.
"While his debut marks a significant moment in his football career, everyone at Theale Green School knows there is much more to come from this exciting young talent.
"We are enormously proud of Maeson and look forward to following his progress as he continues his development in the professional game."