Parts

Overview


Overall, our team used six basic parts and proposed new composite parts to exten their applications. We used Colicin E1 and Colicin E1 immunity protein (BBa_K2922024, BBa_K2922025, proposed by XMU-China in 2019) as genes of interest, both natively derived from E. coli. The promoter used in this project is lactate-inducible ALPaGA ( BBa_K4244000, Wageningen_UR, 2022). It is a codon-optimized wild-type lldRPD promoter (BBa_K2824003 ), which works in anoxia and the presence of glucose. ColE1 is expressed under ALPaGA and double terminator BBa_B0015. Additionally, we added a regulatory gene of the promoter, which encodes for the lldR protein (BBa_K184700 ). For a lldR and ColE1 immunity expression, the P9 promoter is used, BBa_K3606018.

Parts list


Name Nickname Type Description Length(bp)
BBa_K4782006 ALPaGA + RBS+ Colicin E1 Composite Synthesis of ColicinE1 is regulated by the presence of lactate in high concentration in the environment, 1792
BBa_K4782007 P9+lldR + RBS+ ColE1 immunity protein Composite Synthesis of ColicinE1 immunity protein that prevents the cytotoxic effect of ColicinE1 on the producing cells and LldR protein that represses the ColE1synthesis in the absence of lactate are under the control of P9 promoter. 1206
BBa_K4782008 BBa_K4782006+ terminator Composite Synthesis of ColE1 toxin under the control of AlPaGA is controlled by the double terminator B0015 2007
BBa_K4782009 BBa_K4782007+ terminator Composite Synthesis of ColE1 immunity protein and the repressor protein LldR is regulated by double terminator system B0015 1424
BBa_K4782010 pET 9a Basic: Plasmid Backbone In this part, the plasmid that was used as a backbone for our plasmid construction is presented. 4341