- The new temple was sponsored at least in part by Croesus, who founded Lydia's empire and was overlord of Ephesus, and was designed and constructed from around 550 BC by the Cretan architect Chersiphron and his son Metagenes. It was 115 m (377 ft) long and 46 m (151 ft) wide, supposedly the first Greek temple built of marble. Its peripteral columns stood some 13 m (40 ft) high, in double rows that formed a wide ceremonial passage around the cella that housed the goddess's cult image. Thirty-six of these columns were, according to Pliny, decorated by carvings in relief. A new ebony or blackened grapewood cult statue was sculpted by Endoios, and a naiskos to house it was erected east of the open-air altar.
- A rich foundation deposit from this era, also called the "Artemision deposit", yielded more than a thousand items, including what may be the earliest coins made from the silver-gold alloy electrum. The deposit contains some of the earliest inscribed coins, those of Phanes, dated to 625-600 BC from Ephesus, with the legend ΦΑΕΝΟΣ ΕΜΙ ΣΗΜΑ (or similar) (“I am the badge/sign/tomb of Phanes/light”), or just bearing the name ΦΑΝΕΟΣ (“of Phanes”).
- Fragments of bas-relief on the lowest drums of the temple, preserved in the British Museum, show that the enriched columns of the later temple, of which a few survive (illustration below) were versions of this earlier feature. Pliny the Elder, seemingly unaware of the ancient continuity of the sacred site, claims that the new temple's architects chose to build it on marshy ground as a precaution against earthquakes.
- The temple became an important attraction, visited by merchants, kings, and sightseers, many of whom paid homage to Artemis in the form of jewelry and various goods. It also offered sanctuary to those fleeing persecution or punishment, a tradition linked in myth to the Amazons who twice fled there seeking the goddess's protection from punishment, firstly by Dionysus and later, by Heracles.
Electrum coin from Ephesus, 625–600 BC. Stag grazing right, ΦΑΕΝΟΣ ΕΜΙ ΣΕΜΑ (retrograde, “I am the badge/sign/tomb of Phanes/light”).
- In 356 BC, the temple was destroyed in a vainglorious act of arson by a man, Herostratus, who set fire to the wooden roof-beams, seeking fame at any cost; thus the term herostratic fame. For this outrage, the Ephesians sentenced the perpetrator to death and forbade anyone from mentioning his name; but Theopompus later noted it. In Greek and Roman historical tradition, the temple's destruction coincided with the birth of Alexander the Great (around 20/21 July 356 BC). Plutarch remarked that Artemis was too preoccupied with Alexander's delivery to save her burning temple.