Part:BBa_K1031442

J23109-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 at different intensity. BBa_K1031442 is composed of three elements, the constitutive Pc promoter J23109[1], coding sequence of HcaR and terminator B0015[2]. (Fig 1)

Fig 1 Construction of biosensor circuit Ph/J23109-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

Induction ratio of HcaR biosensor library test utilizes different Pc promoters, collocating with RBS (Ribosome Binding Site) at different intensity to control the expression of HcaR. The forth and fifth lines are data for Ph/J23109-HcaR circuit adopting B0034 and B0032 respectively. The dashed box refers to data for Ph/J23109-HcaR biosensor circuit (Fig 2) in collocation with RBS B0034. The data show for Ph/J23109-HcaR circuit, tuning of RBS strength has limited effect.

Fig 2 Induction ratio of HcaR biosensor library test. Horizontal axis stands for different HcaR biosensor with Pc promoters and RBSs 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 Parts' registry. PPA was tested following Test Protocol 1 [http://2013.igem.org/Team:Peking/Team/Notebook/Protocols] as inducer. Vertical axis represents the ON-OFF induction ratio. The dashed box refers to performance for Ph/J23109-HcaR biosensor circuit adopting RBS B0034.