We extensively performed 3A assembly to develop these composite parts.
These parts also include our second nodes and our third nodes for the incoherent feedforward loop design. Keeping in mind the expression of gene and compatibility of plasmids, the first node was placed in an expression vector containing ColE1 origin with chloramphenicol resistance, the second node in the expression vector containing p15A origin with Kanamycin resistance and the third node in the expression vector containing pSC101 origin with Ampicillin resistance. These compatible plasmids were cotransformed in E. coli DH5alpha Z1 strain to realize our project.
Below we have provided the gel electrophoresis results for our final construction of the composite parts via 3A assembly of the various components step-by-step. The observed size of the DNA in the gel matches with the expected size of DNA, as proposed in the design.