Part:BBa_K1031444

J23114-HcaR

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

Characterization

In order to fine-tune the performance of HcaR biosensor, we constructed a library of Pc constitutive promoters and RBSs (Ribosome Binding Sites) at different intensity. BBa_K1031444 is composed of three elements, the constitutive Pc promoter J23114[1], coding sequence of HcaR and terminator B0015[2]. (Fig 1)

Fig 1 Construction of biosensor circuit Ph/J23114-HcaR

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 892
Illegal AgeI site found at 565
1000
COMPATIBLE WITH RFC[1000]

Data shown

We constructed a RBS library of different intensity controlling expression of sfGFP in collocation with Ph/J23114-HcaR biosensor circuit. The last three lines are data for Ph/J23114-HcaR circuit in collocation with RBS B0031, B0032 and B0034 respectively. The dashed box refers to data for Ph/J23114-HcaR circuit adopting RBS B0031(Fig 2), which obtains slight better performance comparing with B0032, B0033 and B0034, indicating tuning for RBS doesnn't obtain obvious effect.

Fig 2 Induction ratio of HcaR biosensor library test. Horizontal axis stands for different HcaR biosensor with Pc promoters of different strength. The expression strength of these constitutive promoters, J23113, J23109, J23114, J23105, J23106 is 21, 106, 256, 623, and 1185, respectively, according to the Partsregistry. Four kinds of aromatics shown with different color intensities, were tested following Test Protocol 1 [http://2013.igem.org/Team:Peking/Team/Notebook/Protocols]. Vertical axis represents the ON-OFF induction ratio. The dashed box refers to performance for Ph/J23114-HcaR biosensor with RBS B0031.