- Miko (? – 1754)
- Haytham Kenway (1754 – 1781)
- Charles Lee (1781 – 1782)
- Ratonhnhaké:ton (1782 – 1783)
- Desmond Miles (2012)
In 1754, the key was in the possession of the Assassin Miko. On the orders of Templar Grand Master Reginald Birch, Haytham Kenway assassinated the man inside the Theatre Royal in London and took the key from him. Upon Haytham's return to the Templar headquarters, Birch told Haytham that the key most likely opened a First Civilization storehouse in the British colonies. As a result, Birch booked Haytham passage to Boston, in order to find it and unlock its contents, and to establish a permanent Templar presence in colonial America.
During 1755, Haytham, with the help of Kaniehtí:io from the Kanien'kehá:ka nation, located the entrance to the Grand Temple, but they were unable to access it, as he did not possess an Apple of Eden to open the first door. Following this, Haytham kept the amulet in his possession until 1781, when he passed it on to his second-in-command, Charles Lee.
In the following year, the Assassin Ratonhnhaké:ton, Haytham's son, killed Charles Lee in Monmouth and took the amulet from him. He later returned to his village, Kanatahséton, where he activated his tribe's Crystal Ball he had used years earlier to communicate with Juno. Juno greeted Ratonhnhaké:ton again, and told him to hide the amulet where none might find it. Following this, he made his way back to the manor at the Davenport Homestead and hid the key in the grave of Connor Davenport, after whom Ratonhnhaké:ton's mentor Achilles Davenport had nicknamed him.
In 2012, Ratonhnhaké:ton's descendant, Desmond Miles, relived his memories inside the Animus and found out that the key was hidden in Connor Davenport's grave. After having retrieved it, Desmond used it to open the inner gateway of the Grand Temple.
- The key contained the Ouroboros symbol, which consists of a snake, or a dragon, eating its own tail. The symbol represents the cyclical nature of many things in the universe, and of things beginning anew. It also represents infinity and a never ending cycle.
- The symbols on the key include the twelve signs of the western zodiac, the Venus symbol, and three other incomplete symbols which are likely the Mars, Moon, and another classical planet symbol.
- Although Ratonhnhaké:ton canonically buried the key, it could still be found with the other mementos of Ratonhnhaké:ton's targets in the basement of the Davenport Homestead.