Significance Insulators are genome sequence elements that help to organize eukaryotic genomes into coherent regulatory domains. Insulators can encode both enhancer-blocking activity, which prevents the interaction between enhancers and promoters located in distinct regulatory domains, and/or chromatin barrier activity that helps to delineate active and repressive chromatin domains. The origins and functional characteristics of insulator sequence elements are important, open questions in molecular biology and genomics. This report provides insight into these questions by demonstrating the origins of a number of human insulator sequences from a family of transposable-element–derived repetitive sequence elements: mammalian-wide interspersed repeats (MIRs). Human MIR-derived insulators are characterized by distinct sequence, expression, and chromatin features that provide clues as to their potential mechanisms of action.