Vibrio cholerae is a pathogen responsible for the deadly pandemic - cholera. The glycans present on the surface of various strains of V. cholerae are considered as potential vaccine candidates. The tetrasaccharide repeating unit (RU) of V. cholerae O43 is decorated with less-explored rare deoxy amino sugars like d-quinosamine and d-viosamine, along with a rare amino acid, N-acetyl-l-allothreonine. Herein, we report a detailed account of the total synthesis of V. cholerae O43 tetrasaccharide RU. In our earlier attempt, while a one-pot assembly of trisaccharide was successful, the final coupling with a fully functionalized d-viosamine donor was low yielding. The successful route involved employing the Fmoc-protected d-viosamine building block as a donor and a late-stage amide bond formation of the tetrasaccharide.