Overhangs of DNA fragments formed by the activity of molecular scissor EcoRI, consists of the following base sequence from 3ʹ to 5ʹ direction.

EcoR I, recognizes the palindromic sequence GAATTC on both the DNA strands and cuts the DNA strands between G and A to form the overhangs at both the ends of the DNA fragment as AATT in 5ʹ to 3ʹ direction or TTAA from 3ʹ to 5ʹ direction.