Several biobricks from the distribution kit were used during our project. These were retrieved from the kit using the protocols provided by iGEM, transformed into E. coli and isolated in elution buffer NE. We also ordered several constructs from IDT (Table 1). These Biobricks were used to create several new biobricks that were relevant to our project. BBa_K2736101 and BBa_K2736103, were created to be used as BRET pair to measure the activity of our customized Tar receptors (BBa_K2736106, BBa_K2736108). To test whether it was possible to switch ligand binding domains of the Tar receptor and two component receptors we created BBa_K2736105, and BBa_K2736109. During our project we proved the functionallity of the parts BBa_K2736101 and BBa_K2736105, which were submitted to the registery.
Table 1: Acquired Biobricks
Name
Biobrick Number
Size
Function
Obtained
RFP
BBa_K516032
828
Red fluorescence
Isolated
Promoter
BBa_K608003
56
Strong promoter with medium ribosome binding site
Isolated
eYFP
BBa_I15017
724
Yellow fluorescence
Isolated
CheZ
BBa_K629003
645
CheZ chemotaxis regulator protein phosphatase for CheY
Isolated
Copper promoter
-
98
Promoter responsive to phosphorylated CusR
Ordered
Custom receptor
BBa_K2736109
1505
Fusion protein of CusS with the Tar ligand binding domain
