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The synagogue, of Italian rite, is today practically unrecognisable from the outside. On the same site since the Ghetto was instituted (1632), the building was damaged in the 1930 earthquake and dismantled and sold a few years later. Its main entrance was on Via delle Scuole, in front of the Spanish synagogue, but it had secondary entrances (partly for security reasons) on Via delle Botteghe, one of which led directly to the section reserved for women.

The hall, designed with a bifocal layout, housed an aron hakodesh surrounded by a semicircular shrine structure formed by six trabeated columns surmounted by a cupola. According to some, the central core – the compartment for the Scrolls of the Law – came from Ascoli Piceno, from where it was brought by the community that was exiled from the papal territories in 1569 and moved as a group to the Duchy of Pesaro and Urbino. Today the furniture belongs to an oratory of the Central Synagogue of Milan, while the surrounding elements are located in Netanya, Israel. On the wall opposite the aron was a raised tevah with canopy, accessed by symmetrical staircases. Today it has partially been relocated to the Istambuli synagogue, in the complex comprising the four Sephardic synagogues of the Old City of Jerusalem.

A third synagogue of Italian rite is documented to have stood in the present-day Via delle Zucchette (formerly Piazza Giudea). With the institution of the ghetto it had to be abandoned because it was the outside the area of reclusion.

Via delle Scuole 26