The four-inch piece of hair was bought by UK collector Paul Fraser at Heritage Auctions in Dallas, Texas.
A German hairdresser kept a piece of Lennon's hair after giving him a trim before the star began filming the 1967 dark comedy, How I Won the War.
Richard Lester's film follows the World War Two misadventures of British troops led by an incompetent commander.
Lennon played Private Gripweed in the film, donning his now signature style round glasses.
"This is the largest lock of John Lennon's hair ever offered at auction and this world record price is a lasting testament to the world's more than 50-year love affair and fascination with Lennon and the Beatles," said Garry Shrum, director of music memorabilia at Heritage.
Also auctioned was a signed glossy head-and-shoulders "before shot" portrait of Lennon, taken shortly before the haircut, which sold for $2,125 (£1,475).
Several other Beatles items were also up for sale at the auction, including a photograph of the band signed by all four members which went for $42,500 (£29,500), and an unused ticket for the Beatles' first US concert in Washington D in 1964, which fetched $30,000 (£20,827).
The large gloss black-and-white photograph by Dezo Hoffmann featured the Beatles performing at the Empire Theatre, Liverpool on 7 December 1963.
The biggest seller was a rare sealed copy of the band's US LP Yesterday and Today, which went for $125,000 (£86,778).
Other sales at the entertainment and music memorabilia auction included a 1980s wristwatch that belonged to Apple co-founder Steve Jobs, which went for $42,500 (£29,500).
A lock of John Lennon's hair that was cut off as he prepared for a film role has sold for $35,000 (£24,298).