According to CNN, the silver Aston Martin DB5 features all the original specifications made by the car manufacturer for the movie, including radar navigation, front machine guns that shoot blanks, a prop ejector seat, a nail spreader, tire shredders, a bullet shield and a revolving number plate.
The vehicle is expected to fetch $5m (£3.1m) to $7m (£4.4m) in a London auction this week.
"It's a once-in-a-lifetime opportunity. I think, unquestionably, it's the most famous car in the world," said Peter Wallman, car specialist at RM Auctions.
The car only had one private owner before going up for auction and has 30,000 miles on its clock. Owner Jerry Lee said that he bought it from Aston Martin directly and has kept the vehicle in his "James Bond room" for the past four decades.
"I drove it for two blocks in London. I think it's a work of art. It's just pure beauty," Lee said.
Proceeds from the sale will go to Lee's charity organisation Jerry Lee Foundation, which works to reduce crime.