Part:BBa_K1031422

Ph(HcaR)-B0034-sfGFP

For detailed information concerning HcaR and Ph promoter, please visit 2013 Peking iGEM Biosensor HcaR

Structure

Fig 1 Structure of Ph promoter regulated by HcaR and chemical pathway.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 113

Construction and data

For the biosensor circuit Ph/HcaR, we created a library of RBS (Ribosome Binding Site) including B0031, B0032 and B0034 for tuning the expression level of reporter gene sfGFP. K1031422 is composed of three elements, Ph promoter activated by HcaR, RBS B0034 and reporter gene sfGFP (Fig 1a). Induction ratio was tested when K1031422 is in collocation with J23106-HcaR, J23109-HcaR, J23113-HcaR and J23114-HcaR, as shown in the dashed boxes (Fig 1b). From data we concluded biosensor circuit Ph/HcaR adopting Pc J23106 and RBS B0034 obtained the optimal performance.

Fig 1 Construction and tunning of reporter circuit Ph-B0034-sfGFP. (a) The orange arrow represents Ph promoter activated by HcaR. The green oval stands for RBS B0034. sfGFP coding sequence is shown with dark blue, while terminator B0015 is in dark red. (b) Test was done following Test Protocol 1. Horizontal axis represents biosensor circuit adopting different combination of Pc promoter and RBS. Vertical axis stands for induction ratio. The dashed boxes refer to data for reporter circuit Pc-B0034-sfGFP in collocation with biosensor circuits J23106-HcaR and J23114-HcaR respectively.