Progress in plant molecular tools has been resulted in the development of gene-targeted and functional marker systems. CAAT box region is a different pattern of nucleotides with a consensus sequence, GGCCAATCT, which situated upstream of the start codon of eukaryote genes and plays an important role during transcription. In the present study, several CAAT box-derived polymorphism (CBDP) primers were used for fingerprinting in mini-core collection of durum wheat (including internationally developed breeding lines and Iranian landraces). Twelve selected primers amplified 98 loci, of which all were polymorphic. The average values of the polymorphism information content (PIC) and resolving power (Rp) were 0.31 and 9.16, respectively, indicating a high level of variability among studied genotypes. Analysis of molecular variance (AMOVA) indicated that 92% of the total variation resided among populations. The values of the percentage polymorphic bands (PPL), the observed (Na) and effective (Ne) number of alleles, Nei’s gene diversity (He) and Shannon’s information index (I) for Iranian landraces were higher than the breeding lines. The Fandendrogram obtained by cluster analysis grouped all individuals into three main clusters. Our results showed a remarkable level of genetic diversity among studied durum wheat, especially among Iranian landraces, which can be interest for future breeding programs. More importantly, the present study also revealed that CBDP technique was efficient and powerful tool to assess genetic diversity in wheat germplasm. Hence, this technique could be employed individually or in combination with other molecular markers to evaluate genetic diversity and relations among different species.