The location in the mouse genome of the 149-base pair MES-1 element, previously isolated by its ability to restore expression to an enhancerless selectable gene, was analyzed. The active moiety of the single-copy MES-1 element is located between the 5' ends of two divergent transcription units, SURF-1 and SURF-2, both of which specify more than one mRNA species by differential splicing. The heterogeneous 5' ends of the SURF transcripts are separated by only 50 to 75 base pairs, and this sequence possesses a high G + C content (65%) and contains neither the TATA and CAAT box motifs normally associated with many highly expressed genes nor the GC box motif (Sp1-binding site) associated with a number of housekeeping genes. Although MES-1 appears to have enhancerlike properties when linked to heterologous genes, its normal genomic location suggests that it functions as a bidirectional promoter. Thus, MES-1 may represent a new class of enhancer-promoter element.