Reagent	Volume for 1x reaction (µl)
DNA vector (~50 ng/µl)	1
PCR amplified fragment-1 (~50 ng/µl)	1
PCR amplified fragment-2 (~50 ng/µl)	1
PCR amplified fragment-3 (~50 ng/µl)	1
PCR amplified fragment-4 (~50 ng/µl)	1
2x Gibson Assembly master Mix	5
Total volume	10