Team:CSIA SouthKorea/design


Since gene parts were already on the partsregistry, we linked them using Gibson assembly. However, our model is different from original plasmids used by Danino et al. We used luxpR only once to bring aiia, GFP, and luxI protein under the promoter. And we used luxpL promoter only once, placing luxR under it.
Since RBS sequence was short, we used gibson assembly to synthesize those four parts.
Then, by connecting those four parts and restriction enzymes, we made luxpR-aiia part to be cutted by EcoRI and SpeI, luxpL-luxR part by PstI and SpeI, and GFP-luxI by XbaI and PstI(illustrate below!).
Plasmid1.jpg Plasmid2.jpg Plasmid3.jpg
Then, by using iGEM standard protocol, in two different types of vectors, PSB1A2 and PSB1K3, we ligated the parts with vectors as shown below.
Ligation1.jpg Ligation2.jpg