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"President Barack Obama was born in the United States. Period," the GOP presidential candidate said in nationally televised comments. "And now we all want to get back to making America strong and great again."
Trump made the acknowledgement at the end of a more than 30 minute event in which he promoted his new Washington hotel and received endorsements from military veterans.
Critics had called his years-long "birther" campaign racially charged.
Trump has previously refused to publicly concede that Obama was born in the United States, despite the president's release of his birth certificate showing he was born in Hawaii. Just Wednesday, during an interview with The Washington Post, Trump refused to say Obama was born in the U.S.
Critics like Democratic presidential rival Hillary Clinton have bashed Trump's years-long campaign against Obama as an attempt to de-legitimize the country's first African-American president. Trump's claims had fueled doubts about his ability to court minority voters, who overwhelmingly have backed Clinton in recent polls.
Despite his statement Friday, Trump did not apologize for stirring the controversy, and Clinton blasted him on Twitter.
Obama told reporters on Friday that he was "shocked" the issue even resurfaced, but added: "I was pretty confident about where I was born."
A Clinton campaign spokesman on Friday called Trump's comments "disgraceful," saying "it was appalling to watch Trump appoint himself the judge of whether the President of the United States is American."
Trump, like his campaign on Thursday, falsely blamed Clinton for starting the so-called "birther" movement in her 2008 presidential primary against Obama. Supporters of Clinton made those claims, but there is no evidence of Clinton or her campaign questioning Obama's birthplace.
Trump also claimed he "finished" the birther theory. His campaign said he did so by compelling Obama to release his birth certificate in 2011.
In tweets since 2011, Trump had suggested he still doubted Obama's birth certificate, even suggesting hackers should find more information about the president.
— CNBC's Katie Little contributed to this report.