Team:Exeter/Basic Part

FimH sfGFP

To easily monitor expression we chose to modify FimH by inserting the super folder green fluorescent protein (sfGFP) (Pedelacq et al 2005). The sfGFP was inserted in three different places in the FimH protein to act as a reporter on the level of expression of the protein and thus giving an indication of its efficiency. The three places of insertion were 22, which is the place where the signal peptide ends in the mannose binding domain, the 225 and 258, which are present in the domain that interacts with FimG.

Name Description Base Pairs
BBa_K2324002 FimH_22_sfGFP 1617
BBa_K2324001 FimH_225_sfGFP 1617
BBa_K2324003 FimH_258_sfGFP 1617

FimH Fusion Proteins

The metal binding proteins were inserted at the 22nd residue of the FimH protein, because that is where the siganl peptide of FimH ends and it is part of the mannose binding domain. Whereas, the 225 and 258 inserts are in the domain that interacts with FimG.

Name Description Base Pairs
BBa_K2324014 FimH_22_His 921
BBa_K2324004 FimH_22_SynMT 1071
BBa_K2324005 FimH_22_MouseMT 1086

Fim Operon

The fim operon consists of six proteins with a native RBS, it was produced by a previous iGEM team from Harvard in 2015.

Name Description Base Pairs
Part:BBa_K1850012 Fim Operon 5744