Using a 5,000-person DNA database from the Cumberland Gap Region of Appalachia, we document the presence of a Sephardic and Ashkenazic Jewish settlement in Central Appalachia. The settlement may have begun as early as the mid-sixteenth century with the Pardo Expedition and been substantially supplemented from the early seventeenth century onward with Jewish colonists from England, Scotland, and Wales. Additional persons found in this mountainous region show DNA origins from Southeastern Europe, North Africa, South Asia, and the Middle East. Thus the region may have served as a refuge for non-white, non-Christian persons arriving in Colonial North America.

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