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 |