GCA_904849725.1 MorexV3_pseudomolecules_assembly Sequence assembly of large and repeat-rich plant genomes has been challenging, requiring substantial compute resources and often several complementary sequencing and mapping approaches. The recent development of fast and accurate long-read sequence assembly by circular consensus sequencing (CCS) on the PacBio platform may greatly increase the scope of plant pan-genome projects. Here, we compare current long-read sequencing platforms regarding their ability to rapidly generate contiguous sequence assemblies in pan-genome studies. The final Morex V3 assembly was constructed the following: As the CCS_Canu assembly outperformed the other assemblies in almost all regards, it was chosen as the primary contig assembly to construct chromosomal pseudomolecules. Pseudomolecule construction proceeded in the three steps: (i) scaffolding the CCS_Canu with Bionano contigs (ii) removal of small redundant sequences (iii) filling gaps in scaffolds with ONT_smartdenovo contigs and (iv) ordering and orienting scaffolds into chromosomal pseudomolecules.<br/><p><b>Comment</b><br/>The construction of the PacBio long-read barley Morex reference assembly proceeded in three steps: (i) scaffolding the CCS_Canu assembly (PRJEB40581) with Bionano contigs (ii) removal of small redundant sequences (iii) filling gaps in scaffolds with ONT_smartdenovo contigs and (iv) ordering and orienting scaffolds into chromosomal pseudomolecules. MorexV3_pseudomolecules_assembly chromosome clone or isolate PacBio RSII Hi-Canu (commit r9818) 25 full 112509 Hordeum vulgare subsp. vulgare SAMEA7384724 PRJEB40589 CAJHDD 01 1H Chromosome 2H Chromosome 3H Chromosome 4H Chromosome 5H Chromosome 6H Chromosome 7H Chromosome WGS_SET_FLATFILE ftp://ftp.ebi.ac.uk/pub/databases/ena/wgs/public/caj/CAJHDD01.dat.gz WGS_SET_FASTA ftp://ftp.ebi.ac.uk/pub/databases/ena/wgs/public/caj/CAJHDD01.fasta.gz total-length 4225577519 ungapped-length 4224251725 n50 610333535 spanned-gaps 162 unspanned-gaps 0 scaffold-count 290 count-contig 452 contig-n50 69630691 contig-L50 15 contig-n75 39657778 contig-n90 19323248 scaf-L50 4 scaf-n75 561794515 scaf-n90 516505932 replicon-count 7 count-non-chromosome-replicon 0 count-alt-loci-units 0 count-regions 0 count-patches 0 ENA-LAST-UPDATED 2026-04-15