Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive disease that threatens to wheat production in China. Breeding resistance cultivars is the effectively control strategy. Wheat line M321 has conferred a high level of all-stage resistance (ASR) to Pst. While the genetic basis of stripe rust resistance in this line has not been deciphered. A doubled haploid (DH) population with 85 lines and an F2 population with 541 individuals were both developed from a cross between a highly susceptible cultivar Superb and M321. The DH population was used to construct genetic map using Wheat55K SNP (single nucleotide polymorphism) chip array. A high-density genetic map contains 16,949 SNP markers and 11 other markers (SSR, KASP and dCAPS) spanning 3222.43 cM across the 21 wheat chromosomes was constructed. Based on the constructed maps, two QTL Yr.nwipb-7BL and Yr.nwipb-7DS for stripe rust resistance was detected across four environments in a 1.85 cM and 2.17 cM intervals on 7B and 7D, respectively. The LOD values of the 7B and 7D QTLs were 6.08 and 7.66, and the additive effects were 0.05 and 0.11, respectively. The additive effect was positive in the two QTLs, indicating that the resistance loci were derived from M321. The phenotypic variation explained (PVE) by the two QTLs was 31.15% and 10.08%, respectively. The Yr.nwipb-7BL was also detected using the bulked segregant RNA-Seq (BSR-Seq) analysis. The closest linked marker was 7B7, with a 0.47 cM genetic distance between it and Yr.nwipb-7BL. Compared with the previously identified resistance genes, Yr.nwipb-7BL could be a new all-stage resistance (ASR) gene against the current epidemic of Pst races in China. Moreover, Yr.nwipb-7DS may be the same locus as the adult-stage resistance gene Yr18.