Part:BBa_K1031017

J23114-B0034-RiboJ-φR73δ

For detailed information concerning Band-pass Filter, please visit 2013 Peking iGEM Band-pass Filter

Characteristic

We constructed this circuit to test the inhibition effect of CⅠ repression circuit. It is composed of constitutive promoter J23114, Ribosome Binding Site B0034，RiboJ ribozyme and transcriptional activator φR73δ. It is used as a component in inhibition circuit to test if CⅠ could inhibit the hybrid promoter.

Fig 1 Left Construction of J23114-φR73δ circuit. The orange arrowhead stands for promoter J23114, the green oval represents RBS B0034, φR73δ shown in brown square. Right Inhibition circuit of Band-pass Filter, consisting of NahR, CⅠ,φR73δ and reporter sfGFP.

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
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Usage in Band-pass Filter

This circuit is an assistant device aiming at testing the inhibition effect of CⅠ protein to hybrid promoter. We propose to adopt φR73δ to initiate constitutive expression of reporter sfGFP, thus construct a background with high fluorescence intensity. When CⅠ protein is expressed, the fluorescence intensity is supposed to go down if the inhibition circuit really works.

Fig 3 K1031017 is designed to construct a high fluorescence background level, which is used to test inhibition of CⅠ protein. Horizontal axis represents 4-MeSaA at different concentration. Vertical axis stands for induction ratio calculated from fluorescence intensity. From Fig 3 it is obvious that CⅠ protein indeed functions as an inhibitor to the hybrid promoter.