The nuclear phosphoprotein cAMP-response-element-binding-protein 1 (CREB1) is involved in the transcriptional regulation of cAMP-inducible genes. We have determined the chromosomal location of the human CREB1 gene by using a cDNA probe. Southern blot analysis of genomic DNA from a panel of mouse-human somatic cell hybrids indicated that the CREB1 gene resides on human chromosome 2. Analysis of a somatic cell hybrid containing a chromosome 2 translocation narrowed assignment of the CREB1 locus to the terminal third of chromosome 2. Finally, in situ hybridization allowed localization of the gene to the 2q32.3-q34 region. This knowledge may prove valuable in molecular analysis of translocations occurring in this region and in exploring the possibility of a role for the CREB1 locus in malignancy.