Aptamers are attractive tools for modulating function of a desired target. In this study, we isolated an RNA aptamer that specifically inhibits transcription of SP6 RNA polymerase. The dissociation constant and 50% inhibitory concentration of the aptamer were estimated 9.5 nM and 24.8 nM, respectively. Doped-SELEX and mutational analysis revealed that the aptamer adopts the structure including two stems, two loops, and 5' single-stranded region. Based on the results, the aptamer could be engineered to circular permutant and binary construct forms without decreasing the activity. The aptamer would be applicable for the construction of expression regulation systems. (C) 2012 Elsevier Inc. All rights reserved.