Difference between revisions of "Part:BBa K2387029"

Revision as of 09:40, 18 October 2017

CpxR-mVenus[1-157] and CpxR-mVenus[158-238] + araC/pBAD promoter

The [http://parts.igem.com/Part:BBa_K2387038 N-terminus of mVenus] and the [http://parts.igem.com/Part:BBa_K2387039 C-terminus of mVenus] are fused to Cpx response regulator [http://parts.igem.com/Part:BBa_K2387002 CpxR] in order to visualize the activation of the Cpx pathway. Upon activation of the Cpx pathway, CpxR gets phosphorylated by E. coli endogenous CpxA after which it can homodimerize. This protein-protein interaction can be visualized using BiFC, hence the fusion of mVenus[1-157] and mVenus[158-238]. The fusion is put under the control of the L-arabinose inducible araC/pBAD promoter. Strong RBS BBa_B0034 is used to regulate transcription.

CpxR-mVenus[1-157] and CpxR-mVenus[158-238] fusions were linked together using a [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] consisting of two times the amino acids GGGGS.

This part is used to visualize the activation of the Cpx pathway via Bimolecular Fluorescence Complementation by using the CpxR-CpxR interaction.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1205
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1144
Illegal XhoI site found at 1283
Illegal XhoI site found at 2498
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 979
Illegal AgeI site found at 1694
Illegal AgeI site found at 2909
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 961

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2387029 short</partinfo>
-The [http://parts.igem.com/Part:BBa_K2387038 N-terminus of mVenus] and the[http://parts.igem.com/Part:BBa_K2387039
+The [http://parts.igem.com/Part:BBa_K2387038 N-terminus of mVenus] and the [http://parts.igem.com/Part:BBa_K2387039 C-terminus of mVenus] are fused to Cpx response regulator [http://parts.igem.com/Part:BBa_K2387002 CpxR] in order to visualize the activation of the Cpx pathway. Upon activation of the Cpx pathway, CpxR gets phosphorylated by <i>E. coli</i> endogenous CpxA after which it can homodimerize. This protein-protein interaction can be visualized using BiFC, hence the fusion of mVenus[1-157] and mVenus[158-238].
-C-terminus of mVenus] are fused to Cpx response regulator [http://parts.igem.com/Part:BBa_K2387002 CpxR] in order to visualize the activation of the Cpx pathway. Upon activation of the Cpx pathway, CpxR gets phosphorylated by <i>E. coli</i> endogenous CpxA after which it can homodimerize. This protein-protein interaction can be visualized using BiFC, hence the fusion of eYFPn[1-154] and eYFPc[155-238].
 The fusion is put under the control of the L-arabinose inducible [https://parts.igem.org/Part:BBa_I0050 araC/pBAD promoter]. Strong RBS [https://parts.igem.org/Part:BBa_B0034 BBa_B0034] is used to regulate transcription.
-CpxR-mVenusn and CpxR-mVenusc fusions were linked together using a [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] consisting of two times the amino acids GGGGS.
+CpxR-mVenus[1-157] and CpxR-mVenus[158-238] fusions were linked together using a [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] consisting of two times the amino acids GGGGS.
 This part is used to visualize the activation of the Cpx pathway via Bimolecular Fluorescence Complementation by using the CpxR-CpxR interaction.