Deer tick virus (DTV) is a genetic variant of Powassan virus (POWV) that circulates in North America in an enzootic cycle involving the blacklegged or "deer tick," Ixodes scapularis, and small rodents such as the white-footed mouse. The number of reported human cases with neuroinvasive disease has increased substantially over the past few years, indicating that POWV may be of increasing public health importance. To this end, we sought to estimate POWV infection rates in questing I. scapularis collected from four health districts in Maine (York, Cumberland, Midcoast, and Central Maine). Infection rates were 1.6%, 1.7%, 0.7%, and 0%, respectively, for adults collected from April to November in 2016. Adults collected in October and November in 2017 from York and Cumberland counties had slightly higher rates of 2.3% and 3.5%, respectively. There was no difference in the number of males verses the number of females infected. All positive samples were of the DTV (lineage II) variant. Phylogenetic analysis was performed on 8 of the 15 DTV sequences obtained in 2016. Deer tick virus from the coastal regions were genetically similar and clustered with virus strains isolated from I. scapularis from New York State and Bridgeport, CT. The two inland viruses were genetically nearly identical and grouped with viruses from Massachusetts, Connecticut, and New York. These results are the first reported infection rates and sequences for POWV in questing ticks collected in Maine and will provide a reference point for future POWV studies.