•Wheat stem rust, caused by Puccinia graminis Pers. f. sp. tritici (Pgt), is a devastating fungal disease threatening global wheat production. Here, we report the identification of stem rust resistance gene Sr60, a race-specific gene from diploid wheat Triticum monococcum L. that encodes a protein with two putative kinase domains. This gene, designated WHEAT TANDEM KINASE 2 (WTK2), confers intermediate levels of resistance to Pgt. •WTK2 was identified by map-based cloning and validated by transformation of a ~10-kb genomic sequence including WTK2 into susceptible common wheat variety Fielder (T. aestivum L.). •Transformation of Fielder with WTK2 was sufficient to confer Pgt resistance. Sr60 transcripts were transiently upregulated one day after Pgt inoculation, but not in mock-inoculated plants. The upregulation of Sr60 was associated with stable upregulation of several pathogenesis-related genes. •The Sr60-resistant haplotype found in T. monococcum was not found in polyploid wheat, suggesting an opportunity to introduce a novel resistance gene. We successfully introgressed Sr60 into hexaploid wheat and developed a diagnostic molecular marker to accelerate its deployment and pyramiding with other resistance genes. The cloned Sr60 can also be a useful component of transgenic cassettes including other resistance genes with complementary resistance profiles. This article is protected by copyright. All rights reserved.