The chopper Peter Fonda rode into film history in Easy Rider (1969) is set to cross the block at Profiles In History in Los Angeles on October 18.
It's expected to make $1m-1.2m.
The iconic bike, a customised "Captain America" Harley-Davidson featuring a stars and stripes fuel tank, is one of four used in filming.
It's shown on fire by the side of the road in the final scene. The other three bikes were stolen shortly before the film wrapped.
Fonda later restored the bike and presented it to production assistant Dan Haggerty, who donated it to the National Motorcycle Museum.
Michael Eisenberg, a business partner of Fonda, is consigning the piece of film history after acquiring it from the museum. The profits from the sale will be donated to the American Humane Association.
Easy Rider was one of the defining films of its era, directed by and starring Fonda and Dennis Hopper as two hippies travelling from LA to New Orleans, who ride through the fallout of the 1960s.
The production was famously fraught (Fonda and Hopper were high for the majority of filming) but somehow they managed to pull together a cogent narrative.
The success of the movie went on to kick-start the new wave of American filmmaking as studios started to take risks on edgy young filmmakers.
The US flag patch Fonda sported in the film auctioned for $89,625 in 2007.
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