The nucleotide (nt) sequence of the araBAD operon of Escherichia coli B/r has been determined. The nt sequence predicts a transcript of about 4250 nt. The coding regions of araB, araA and araD genes were identified by partial amino acid (aa) sequences of the purified proteins and their cleavage products. We have deduced that the polypeptides encoded by the araBAD operon consist of 566 (araB), 500 (araA) and 231 (araD) aa residues. The operon contains a leader sequence of 27 nt. The intergenic region between araB and araA is 10 bp in length, while the araA and araD genes are separated by 283 bp consisting primarily of six REP sequences. Comparison of the E. coli sequence with that of Salmonella typhimurium [Lin et al., Gene 34 (1985) 111-122; 123-128; 129-134] shows that, while considerable divergence in nt sequence has occurred, the aa sequences are largely conserved. The most pronounced difference, found in the araD gene, is the deletion of a single nt in the S. typhimurium sequence.