AbstractThe genome ofHalobacteriumstrain 63‐R2 was recently reported and provides the opportunity to resolve long‐standing issues regarding the source of two widely used model strains ofHalobacterium salinarum, NRC‐1 and R1. Strain 63‐R2 was isolated in 1934 from a salted buffalo hide (epithet “cutirubra”), along with another strain from a salted cow hide (91‐R6T, epithet “salinaria,” the type strain ofHbt. salinarum). Both strains belong to the same species according to genome‐based taxonomy analysis (TYGS), with chromosome sequences showing 99.64% identity over 1.85 Mb. The chromosome of strain 63‐R2 is 99.99% identical to the two laboratory strains NRC‐1 and R1, with only five indels, excluding the mobilome. The two reported plasmids of strain 63‐R2 share their architecture with plasmids of strain R1 (pHcu43/pHS4, 99.89% identity; pHcu235/pHS3, 100.0% identity). We detected and assembled additional plasmids using PacBio reads deposited at the SRA database, further corroborating that strain differences are minimal. One plasmid, pHcu190 (190,816 bp) corresponds to pHS1 (strain R1) but is even more similar in architecture to pNRC100 (strain NRC‐1). Another plasmid, pHcu229, assembled partially and completed in silico (229,124 bp), shares most of its architecture with pHS2 (strain R1). In deviating regions, it corresponds to pNRC200 (strain NRC‐1). Further architectural differences between the laboratory strain plasmids are not unique, but are present in strain 63‐R2, which contains characteristics from both of them. Based on these observations, it is proposed that the early twentieth‐century isolate 63‐R2 is the immediate ancestor of the twin laboratory strains NRC‐1 and R1.