Figure 1

Construction outline of the MCP tac s promoter clusters. Fragment 5CPtac s with the flanking sequence was amplified by PCR with p5TG as the template. Fragment 1 was generated by digesting fragment 5CPtac s with BamH I. Fragment 2 was digested from fragment 5CPtac s with BamH I and Hind III. Fragment 3 was linearized from the plasmid p5TG with Hind III. Then, the three fragments were assembled together under the action of T5 exonuclease, Phusion DNA polymerase and Taq DNA ligase in the isothermal process.