In this paper, we introduce an anti-adhesion poly(dimethylsiloxane) (PDMS)-coated flexible mold and its applications for room-temperature imprint lithography. The flexible mold is fabricated using an ultraviolet-curable prepolymer on a flexible substrate, and its surface is passivated with a thin layer of PDMS to impart an anti-adhesion property. The highly flexible mold enables conformal contact with a substrate on which a low-viscosity polymer resist is spin-cast in a thin layer. Large-area imprinting is then realized at room temperature under significantly reduced pressure. The mold was durable even after repetitive imprinting of over 200 times. Also, we show a double imprinting on the substrate with a PDMS-coated replica polymeric mold having 500 nm line patterns. This enables the formation of matrix patterns with varying feature heights in less than 7 min.