To promote the film Goldfinger, the two DB5s were showcased at the 1964 New York World's Fair, and it was dubbed "the most famous car in the world", and subsequently sales of the car rose.

In January 2006, one of these was auctioned in Arizona; the same car was originally bought in 1970 from the owner, Sir Anthony Bamford, by a Tennessee museum owner. A car, mainly used for promoting the movie, is now located in the Louwman Museum, Netherlands. The first DB5 prototype used in Goldfinger with the chassis number DP/216/1 was later stripped of its weaponry and gadgetry by Aston Martin and then resold. It was then retrofitted by subsequent owners with nonoriginal weaponry. It later appeared in the film The Cannonball Run (1981), in which it was driven by Roger Moore.

The Chassis DP/216/1 DB5 was stolen in 1997 from its last owner in Florida and is currently still missing.

Since the film, the DB5 has become closely associated with the James Bond franchise and is considered to be the quintessential vehicle of the character. As such, it has reappeared in subsequent films over the years. Within the universe of James Bond, the same car (registration BMT 216A) was used again in the following film, Thunderball, a year later.

A different Aston Martin DB5 (registration BMT 214A) was used in the 1995 Bond film GoldenEye, in which the car is Bond's personal vehicle and has no gadgets. Although it did come equipped with a champagne cooler in the arm rest and a fax machine. Three different DB5s were used for filming. This same car briefly reappeared in the next film, Tomorrow Never Dies (1997), and was set to make a cameo appearance in the Scotland-set scenes in The World Is Not Enough (1999), but these were cut in the final edit. Yet another DB5 (registration 56526) appeared in Casino Royale (2006), this time owned by the villain, Alex Dimitrios. This car has Bahamian number plates and left-hand drive (where the previous British versions had been right-hand drive). In the film Bond wins the car from Dimitrios after beating him in a card game.

The Goldeneye DB5 currently resides at The London Film Museum in Covent Garden. Of the three cars used in the production, one car was filmed for the static ocean side shot, one's participation is unknown, and DB5/1885/R was driven in the chase scene against the Ferrari in Monte Carlo. DB5/1885/R was bought at a Christies auction in 2001 and entered the Guinness Book of Records that year for the highest paid for a Bond memorabilia. It was first exhibited at the Bond exhibition at Beaulieu before moving to its new home in The London Film Museum.

Another silver-birch DB5 with the original registration BMT 216A was used in the 23rd James Bond film, Skyfall, during the 50th anniversary of the release of the first James Bond film Dr. No (1962). It is unclear if this car is meant to be the same one Bond won in Casino Royale, refitted with British number plates, right-hand drive, and gadgets. The car is destroyed in the film's climactic finale. It is seen again in Spectre (2015), firstly in Q's underground workshop in various stages of rebuild, and at the film's ending, fully rebuilt, with Bond driving away with it.

On 1 June 2010, RM Auctions announced the upcoming auction of a DB5 used in both Goldfinger and Thunderball. The owner (Jerry Lee, president/owner of WBEB Radio in Philadelphia, Pennsylvania) originally bought the car from the Aston Martin company in 1969. At the auction, the DB5 was sold for £2.6 million. (£2.912m with commission) equates to $4.6m.

Aston Martin announced in August 2018 that it plans to build 25 replicas of the DB5 as seen in Goldfinger, including some of the gadgets seen in the film, each selling for about GB£2.75 million.