The Lions will play in front of 16,500 spectators in the match with Japan at BT Murrayfield Stadium on Saturday 26 June.
The curtain-raiser to the Lions Series in South Africa will host the partial capacity crowd following consultation with the Scottish Government and the application of agreed social distancing criteria to ensure fans can attend safely.
It will be the first time supporters will be able to attend the home of Scottish Rugby to watch an international match since March 2020.
"We're absolutely delighted to be able to confirm today’s news," said Lions managing director Ben Calveley.
"It will be brilliant to see a passionate crowd back at Murrayfield for what promises to be a superb game of rugby before we embark on the Tour to South Africa.
"I am sure it will give the entire touring party an enormous boost to play this fixture in front of Lions fans.
"We are very grateful to the Scottish Government for its commitment and support to ensure this fixture would be seen by a live audience."
Meanwhile Twickenham will host crowds of 10,000 for both of this weekend's European finals.
Friday night's Challenge Cup clash between Leicester and Montpellier takes place the day before La Rochelle and Toulouse collide at the same venue in the Heineken Champions Cup showpiece.
It will be the first time spectators have been allowed into the ground since the Autumn Nations Cup final between England and France in December.
"The fans really do make the game and we hope everyone is looking forward to attending live rugby again at Twickenham," Rugby Football Union chief executive Bill Sweeney said.