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The San Francisco 49ers will be the opponents for the Jacksonville Jaguars at Wembley Stadium next year, the league announced on Thursday.
The Jaguars have already been confirmed as the 'home' team for the Wembley fixture in each year between 2013 and 2016.
Jaguars owner Shad Khan confirmed the match-up against the 49ers while attending the Leaders in Sport conference in London.
"Everyone associated with the Jacksonville Jaguars is very much looking forward to our four home games in London through 2016, and we're particularly thrilled and honoured that our first opponent will be the San Francisco 49ers," Khan said.
"This is a unique and important opportunity for the Jaguars franchise, as well as for our fans and community. To share this stage in our first year at Wembley with a rising power like the 49ers will bring additional attention and respect to what we're trying to achieve as a franchise.
We can't wait to see our global vision for the Jaguars become reality next season."
The 49ers were the 'home' team at Wembley in 2010, beating the Denver Broncos 24-16.
"The 49ers are excited and privileged to once again serve as ambassadors for the NFL and the game of football internationally," said 49ers chief executive officer Jed York.
"Our 2010 experience in London was tremendous. The atmosphere at Wembley Stadium on game day was electric. The ability for us to share 49ers football with our faithful fans internationally is a treat for everyone in our organisation."
This year's Wembley game will see the St Louis Rams play the New England Patriots on October 28.
The NFL has hosted one game at Wembley Stadium annually since 2007, and the prospect of hosting multiple games per season has been under discussion for several years.
The league's chief marketing officer Mark Waller said that work regarding a possible second game in the UK - either at Wembley or elsewhere - in 2013 was ongoing.
"We'll meet with ownership next week and we'll take them through an analysis of what the first five years have done and, as a result of that, what the potential is if we were to add in a second game," he said.
"So, it's up for review with our ownership and, the way it works, we get feedback from our ownership on level of interest, is there a specific commitment? is there more work to be done?
"So there's no fixed determination next week as to whether we do it or not. But, definitely, we've worked hard to build up the case of why we think it's a logical next step."
Waller said the final deadline for a decision would be before the season schedule is drawn up in April, but he would expect an answer either way before then.
"We would probably be reluctant to make it that late because there's a whole logistics infrastructure, ticket sales - there's a whole business model that goes with it," he said.