Late in 1606, English colonists set sail with a charter from the London Company to establish a colony in the New World. The fleet consisted of the ships Susan Constant, Discovery, and Godspeed, all under the leadership of Captain Christopher Newport. They made a particularly long voyage of four months, including a stop in the Canary Islands, in Spain, and subsequently Puerto Rico, and finally departed for the American mainland on April 10, 1607. The expedition made landfall on April 26, 1607, at a place which they named Cape Henry. Under orders to select a more secure location, they set about exploring what is now Hampton Roads and an outlet to the Chesapeake Bay which they named the James River in honor of King James I of England. Captain Edward Maria Wingfield was elected president of the governing council on April 25, 1607. On May 14, he selected a piece of land on a large peninsula some 40 miles (64 km) inland from the Atlantic Ocean as a prime location for a fortified settlement. The river channel was a defensible strategic point due to a curve in the river, and it was close to the land, making it navigable and offering enough land for piers or wharves to be built in the future. Perhaps the most favorable fact about the location was that it was uninhabited because the leaders of the nearby indigenous nations considered the site too poor and remote for agriculture. The island was swampy and isolated, and it offered limited space, was plagued by mosquitoes, and afforded only brackish tidal river water unsuitable for drinking.

The colonists, the first group of whom had originally arrived on May 13, 1607, had never planned to grow all of their own food. Their plans depended upon trade with the local Powhatan to supply them with food between the arrivals of periodic supply ships from England. Lack of access to water and a relatively dry rain season crippled the agricultural production of the colonists. Also, the water that the colonists drank was brackish and potable for only half of the year. A fleet from England, damaged by a hurricane, arrived months behind schedule with new colonists, but without expected food supplies. There is scientific evidence that the settlers at Jamestown had turned to cannibalism during the starving time.

On June 7, 1610, the survivors boarded ships, abandoned the colony site, and sailed towards the Chesapeake Bay. There, another supply convoy with new supplies, headed by newly appointed governor Francis West, intercepted them on the lower James River and returned them to Jamestown. Within a few years, the commercialization of tobacco by John Rolfe secured the settlement's long-term economic prosperity.