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(a)The use of linkers to create tailor-made ends on cloning fragments.Synthetic oligonucleotide duplexes whose sequences represent EcoRI restriction sites are blunt-end ligated to a DNA molecule using T4DNA ligase.Note that the ligation reaction can add multiple linkers on each end of the blunt-ended DNA. EcoRI digestion removes all but the terminal one,leaving the desired 5’-overhangs.(b)cloning vectors often have polylinkers consisting of a multiple array of restriction sites at their coning sites, so restriction fragments generated by a variety of endonucleases can by incorporated into the vector.Nore that the polylinker is engineered not only to have multiple restriction sites bur also to have an uninterrupted sequence of codons,so this region of the vector has the potential for translation into protein.The sequence shown is the coning site for the vectors M13mp7 and pUC7;the colored amino acid residues are contiguous with the coding sequence of the lacZgene carried by this vector(see Figure 13.18)