MYB98is a key regulator of the genetic network behind pollen tube attraction toward the female gametophyte.MYB98is specifically expressed in the synergid cells (SCs), a female gametophyte component cells specialized for pollen tube attraction. However, it had not been clear how exactlyMYB98achieves this specific expression pattern. In the current study, we have determined that a normal SC-specific expression ofMYB98is dependent on a 16-bp-longcis-regulatory element, CATTTACACATTAAAA, freshly named as the “Synergid-specificActivationElement ofMYB98” (SaeM). An 84 bp fragment harboringSaeMin the middle was sufficient to drive exclusively SC-specific expression. The element was present in a significantly large proportion of SC-specific gene promoters and in the promoter ofMYB98homologous genes in the Brassicaceae (pMYB98s). Significance of such family-wideSaeM-likeelement conservation in exclusive SC-specific expression was confirmed by the Arabidopsis-like activation feature ofBrassica oleracea–derivedpMYB98and absence of such feature ofpMYB98derived from a non-Brassicaceae memberPrunus persica. Additionally, the yeast-one-hybrid assay showed that theSaeMcan be recognized by ANTHOCYANINLESS2 (ANL2) and DAP-seq data further suggested for additional three ANL2 homologs targeting the similarcis-element. Overall, our study has concluded thatSaeMplays a crucial role in driving exclusively SC-specific expression ofMYB98and strongly suggests for the involvement of ANL2 and its homologs in its dynamic regulationin planta. Future study on the transcription factors is expected to shed more light on the mechanism behind the process.