I am trying to conduct cloning using Gibson assembly using the Takara Master Mix of three inserts of sizes 81bp, 500bp and 600bp and vector of size 5kb. The primers were designed using the Takara primer design tool and the ratio of the inserts and vector were added after calculating their molar concentration following the standard protocol. Transformation was done in chemically competent NEB5a cells. I am getting colonies on my ampicillin selected plates but none of them are positive. I did gel purification of the digested plasmid before going for the ligation. Any suggestions of where I maybe going wrong will be helpful.