The promoter of the 780 gene of T-right [Thomashow, M., Nutter, R., Montoya, A., Gordon, M. & Nester, E. (1980) Cell 19, 729-739] from Agrobacterium tumefaciens Ti plasmid (pTi15955) was shown to contain an upstream cis-acting element (activator) having enhancer-like properties. To characterize the properties of this promoter element, it was placed in both polarities, upstream and downstream of a Delta-37 deletion mutant of the 780 gene. The Delta-37 deletion contains the entire 780 gene with the 5' flanking sequences deleted upstream of TATA to -37. The effect of the activator on in vivo transcriptional activity was assessed by S1 nuclease mapping utilizing a homologous reference gene as an internal standard. Transcript levels from sunflower crown gall tumors were between 127% and 90% of the wild-type 780 gene depending on the polarity of the activator element when placed directly upstream to the 780 gene Delta-37 promoter. Repositioning the activator element 613 base pairs further upstream increased transcription by 2-fold regardless of polarity. However, the activator element did not promote transcription when placed in either polarity approximately 200 base pairs downstream of the poly(A) addition site. Upstream promoter fragments (TATA-distal) from the octopine synthase (ocs) and agropine synthase (ags) genes were also tested for activation of the Delta-37 construction. The ocs sequences activated transcription of the Delta-37 deletion to 14% of wild-type levels when placed upstream in the B (reverse) orientation. All other constructions with the ocs and ags sequences showed no enhancement of promoter activity.