The promoter of the gua operon has been located by transcript mapping using primer extension with reverse transcriptase. The surrounding nucleotide sequence has features characteristic of promoters under stringent and growth-rate-dependent regulation, namely a GC-rich discriminator next to the -10 hexamer, an upstream AT-rich sequence (the UP element) and potential FIS-binding sites. Transcriptional activity of the gua promoter was examined using transcriptional fusions to lacZ placed at a single chromosomal location. Expression from gua was reduced under stringent conditions in vivo, and varied with growth rate. Growth-rate control was independent of guanine-mediated repression. A fusion in which the GC-rich discriminator was mutated by insertion of an AT-rich oligonucleotide was used to demonstrate the importance of this region in control. Both stringent and growth-rate-dependent controls were abolished by the mutation. Other potential regulatory signals in the vicinity of the gua promoter are a pur operator (binding site for the PurR repressor), a gua operator, a DnaA-binding site and a CRP/FNR-binding sequence. The gua promoter lies back-to-back with the promoter for xseA (exonuclease VII), the two promoters being separated by only 20 bp.