Continuous paleoenvironmental records covering the period prior to the Last Glacial Maximum in northeastern Beringia are sparse. This study presents a multi-proxy analysis of a 35-m-high yedoma exposure located on the right bank of the Itkillik River in Alaska. The exposure accumulated over 39 thousand years (kyr) during the Middle Wisconsinan Interstadial and the Late Wisconsinan glacial advance. We identified five stratigraphic units based on pollen, carbon and ice content, and isotopic composition (δ18O) of the sediments. Middle Wisconsinan climate in northern Alaska promoted peat accumulation prior to 33.6 cal kyr BP (calibrated kyr before present). Reconstructed July air temperatures were 1–2 °C lower than current at 34.8 cal kyr BP, consistent with the timing of the interstadial climatic optimum in interior Alaska and Yukon. Colder (by up to 4 °C) and drier conditions characterize the transition from interstadial to glacial conditions between 33.6 and 29.8 cal kyr BP. Late Wisconsinan (29.8–17.9 cal kyr BP) July air temperatures were 2–3 °C lower than today, with grassland vegetation dominated by Poaceae, Artemisia and forbs, in contrast to the modern Cyperaceae dominance. Moister and warmer environmental conditions after 17.9 cal kyr BP correspond to the Late Glacial to Early Holocene interval.