We have previously reported the identification and isolation by mRNA selection/translation of a recombinant clone containing 80% of the human vimentin gene sequence [Lilienbaum et al., EMBO J. 5 (1986) 2809-2814]. We present here the nucleotide sequence of this genomic clone including the 3' untranslated region. To complete the coding sequence, we have isolated cDNA recombinant clones (1.1 kb) of vimentin from human libraries constructed in lambda gt11. Comparison of the coding sequence between human and hamster shows 90% homology at the nucleotide level and four differences out of 353 amino acid residues, as deduced from the nucleotide sequences. In addition to the extensive homology previously reported between the coding sequences of hamster and human vimentin genes [Ferrari et al., Mol. Cell. Biol. 6 (1986) 3614-3620], we observed that the positions of the noncoding regions are also conserved and that the 3' nontranslated region includes two canonic poly(A) signals. Hybridization of the clones to mRNA from different mammalian sources revealed a single species of 2 kb and confirmed that the length of the untranslated and coding sequences are conserved. Quantitative estimations of the mRNA levels in mammalian cells and tissues of various origins are consistent with transcriptional regulation.