A complete genetic linkage map of the soybean, in which sequence-based (SB) genetic markers are evenly distributed genomewide, was constructed from an F(12) population composed of 113 recombinant inbred lines derived from an interspecific cross involving Korean genotypes Hwangkeum and IT182932. Several approaches were employed for the development of 112 novel SB markers targeting both the gaps and the ends of the linkage groups (LGs). The resultant map harbored 20 well-resolved LGs presumed to correspond to the 20 pairs of soybean chromosomes. The map allowed us to identify the important chromosomal structures that were not observed in the integrated genetic maps, to identify the new potentially gene-rich regions, to detect segregation distortion regions within the whole genome, and to extend the ends of the LGs. The results will facilitate the further discovery of agronomically relevant genetic loci in the heretofore neglected chromosomal regions and should also provide some important links between the soybean genetic, physical, and genome sequence maps in the regions.