Ranunculus sceleratus(family: Ranunculaceae) is a medicinally and economically important plant; however, gaps in taxonomic and species identification limit its practical applicability. This study aimed to sequence the chloroplast genome ofR. sceleratusfrom Republic of Korea. Chloroplast sequences were compared and analyzed amongRanunculusspecies. The chloroplast genome was assembled from Illumina HiSeq 2500 sequencing raw data. The genome was 156,329 bp and had a typical quadripartite structure comprising a small single-copy region, a large single-copy region, and two inverted repeats. Fifty-three simple sequence repeats were identified in the four quadrant structural regions. The region between thendhCandtrnV-UACgenes could be useful as a genetic marker to distinguish betweenR. sceleratuspopulations from Republic of Korea and China. TheRanunculusspecies formed a single lineage. To differentiate betweenRanunculusspecies, we identified 16 hotspot regions and confirmed their potential using specific barcodes based on phylogenetic tree and BLAST-based analyses. ThendhE,ndhF,rpl23,atpF,rps4, andrpoAgenes had a high posterior probability of codon sites in positive selection, while the amino acid site varied betweenRanunculusspecies and other genera. Comparison of theRanunculusgenomes provides useful information regarding species identification and evolution that could guide future phylogenetic analyses.