Here we release a chromosome-scale assembly of a Japanese elite malting barley cultivar ‘Haruna Nijo’. A series of genetic and genomic resources have been developed for this genotype and provided by NBRP. The assembly has 4.28Gbp with 18.9Mbp of N50 in size. The pseudomolecule assembly was characterized by transcript evidences of gene projection derived from the reference genome and de novo gene annotation by published full-length cDNA sequences and RNA-seq data of ‘Haruna Nijo’. ‘Haruna Nijo’ has been used for map-based cloning with its BAC library. Since more genes of interest have been identified in this genotype, releases of assembly with detailed genomic information may accelerate the gene identification and utilization in this genotype.