Difference between revisions of "Part:BBa K3132100"

 
 
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<partinfo>BBa_K3132100 short</partinfo>
 
<partinfo>BBa_K3132100 short</partinfo>
  
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This part (4*UAS MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132004) and 4*GAL4 binding sites (fudan2017 BBa_K2446036). 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 and the VP64 domain can activate the expression of 4*UAS MinimalCMV promoter. The corresponding SynTF of UAS_minimalCMV is Gal4-VP64 (BBa_K).  
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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>).
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==Characterization of UAS_MinimalCMV Promotor: ==
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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.
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[[File:T--SMMU-China--UAS_MinimalCMV Promotor.png| 600px|thumb|center|]]
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==References==
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<p>[1] L. Morsut et al., Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Cell 164, 780--791 (2016).</p>
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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.

T--SMMU-China--UAS MinimalCMV Promotor.png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]