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Shortly after completing his apprenticeship under Andrea del Verrocchio, Leonardo established his own workshop in the center of Florence. There, he created commissions for the local townspeople, one of whom was Maria Auditore da Firenze. Through her, Leonardo became acquainted with her son, Ezio, who would frequent his different workshops in later years.
Aside from painting and design, Leonardo also studied human anatomy. He stored and dissected bodies in his workshop, which the city often sent him for research. Because of this, Ezio was once able to hide a guard he had killed in Leonardo's residence.
This studio was where Leonardo first received and decoded a page of Altaïr Ibn-La'Ahad's Codex, with which he repaired the Hidden Blade. Through other pages, he also constructed upgrades for it, including the second blade and an adaption that delivered poison.
Leonardo also allowed Ezio to practice with his Hidden Blade in the courtyard next to the workshop; instructing his assistant at the time, Vincenzo, to set up dummies for him to use.
After receiving a commission from a Venetian noble in 1480, Leonardo left Florence and relocated his workshop to Venice.
- "And now, I present to you, your workshop, Ser da Vinci! We spared no expense in its design! You'll see it is perfect; as if you never left Firenze!"
- ―Alvise da Vilandino, 1481.[src]
Upon his arrival and subsequent tour of the city by Alvise da Vilandino, Leonardo was introduced to his new workshop. Alvise commented that it would be "as if he never left Firenze," and indeed, the building and interior were nearly identical to his workshop in Florence.
Leonardo continued to decode Codex pages for Ezio in this workshop, where he built the Hidden Gun upgrade of the Hidden Blade. As well as this, he also improved the design of his Flying Machine, which allowed Ezio to fly over Venice to the Palazzo Ducale.
During 1488, after retrieving an Apple of Eden from Rodrigo Borgia, Ezio took it to Leonardo's workshop. There, he, Leonardo, Mario Auditore, and Niccolò Machiavelli attempted to discover its purpose.
Despite departing from Venice years later, it was rumored that Leonardo used his workshop there as a secret retreat, returning to it repeatedly during his life.
In 1499, after being forcibly recruited by the Papal Captain General Cesare Borgia, Leonardo began to design and create weapons and war machines for the Borgia forces. As such, by 1500, he had relocated his workshop to Rome.
He remained there even after Cesare's fall from power, and in 1506, Leonardo spent much of his time in his workshop studying Pythagoras and the Pythagorean temple, as well as working on his latest painting, the Mona Lisa.
During that same year, Leonardo was kidnapped from his workshop by Hermeticists, who also ransacked the building's room in search of his map to the temple of Pythagoras. He managed to leave a clue for Ezio to find him by writing on the floor of the workshop.
During their attempts to find the artist, Ezio and Leonardo's assistant Salaì used his workshop to store the da Vinci paintings that they recovered, as well as searching them for clues with Ezio's hidden talent.
- The memory "Ace Up My Sleeve" in Assassin's Creed II was the first time that Ezio entered one of Leonardo's workshops.
- In Assassin's Creed: Brotherhood, Ezio was able to enter the workshop during the Cristina memories that took place in Florence.