Difference between revisions of "Part:BBa K3132100"
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<partinfo>BBa_K3132100 short</partinfo> | <partinfo>BBa_K3132100 short</partinfo> | ||
| − | This part (4*UAS MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132004) and 4*GAL4 binding sites (BBa_K3132014). We inset the 4*Gal4 binding sites before the minimalCMV promoter. MinimalCMV promoter is a promoter from the vector pcDNA3.1 with very strong transcriptional activity. In this way, we can use our synthetic transcription factors (SynTFs) which is a fusing protein Gal4_DBD_(G4S) linker_NLS_VP64. The Gal4-DBD can bind to the 4*Gal4 binding sites specifically[1] and the VP64 domain can initiate transcription by activating the 4*UAS MinimalCMV promoter. The corresponding SynTF of UAS_minimalCMV is Gal4-VP64 (BBa_K3132000). | + | This part (4*UAS MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (<partinfo>BBa_K3132004</partinfo>) and 4*GAL4 binding sites (<partinfo>BBa_K3132014</partinfo>). We inset the 4*Gal4 binding sites before the minimalCMV promoter. MinimalCMV promoter is a promoter from the vector pcDNA3.1 with very strong transcriptional activity. In this way, we can use our synthetic transcription factors (SynTFs) which is a fusing protein Gal4_DBD_(G4S) linker_NLS_VP64. The Gal4-DBD can bind to the 4*Gal4 binding sites specifically[[#References|[1]]] and the VP64 domain can initiate transcription by activating the 4*UAS MinimalCMV promoter. The corresponding SynTF of UAS_minimalCMV is Gal4-VP64 (<partinfo>BBa_K3132000</partinfo>). |
==Characterization of UAS_MinimalCMV Promotor: == | ==Characterization of UAS_MinimalCMV Promotor: == | ||
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| + | <!-- Add more about the biology of this part here | ||
| + | ===Usage and Biology=== | ||
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Latest revision as of 15:54, 21 October 2019
UAS_MinimalCMV Promotor
This part (4*UAS MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132004) and 4*GAL4 binding sites (BBa_K3132014). We inset the 4*Gal4 binding sites before the minimalCMV promoter. MinimalCMV promoter is a promoter from the vector pcDNA3.1 with very strong transcriptional activity. In this way, we can use our synthetic transcription factors (SynTFs) which is a fusing protein Gal4_DBD_(G4S) linker_NLS_VP64. The Gal4-DBD can bind to the 4*Gal4 binding sites specifically[1] and the VP64 domain can initiate transcription by activating the 4*UAS MinimalCMV promoter. The corresponding SynTF of UAS_minimalCMV is Gal4-VP64 (BBa_K3132000).
Characterization of UAS_MinimalCMV Promotor:
We used fluorescence protein mCherry to verify the binding of Gal4-DBD to 4*UAS(Gal4 binding sites), and the results are as follows. We inset mCherry behind 4*UAS and in general the fluorescence intensity is very low; when Gal4 is added, the fluorescence intensity significantly increased which verifies the binding of Gal4-DBD to 4*UAS.
References
[1] L. Morsut et al., Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Cell 164, 780--791 (2016).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]