Difference between revisions of "Part:BBa K3447005"

 
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T7 promoter is a very specific promoter which is transcribed only by specific T7 RNA polymerase which can activate downstream gene expression after being combined.
 
  
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T7 promoter is a very specific promoter which is transcribed only by specific T7 RNA polymerase which can activate downstream gene expression after being combined.<br><br>
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===Usage and Biology===
 
===Usage and Biology===
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In our project, we divided T7 RNA polymerase into two parts, respectively called T7NT (链接) and T7CT (链接) and attached on the nMag and pMag. Under blue light irradiation, these two parts undergo specific blue light dependent heterodimerization and assemble into complete T7rna polymerase, which acts on PT7 promoter and activates downstream gene expression. And the process is reversible, that is, when blue light is stopped, the dimerized T7RNA polymerase de-dimers and loses its function.<br><br>
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The <i>kanR</i> gene is a relatively common type of resistance gene in molecular cloning, and it is also very common in commercial plasmids. In the process of constructing plasmids, we will choose kanamycin-resistant vectors, so we no longer too much detail on its mechanism of action.<br><br>
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[[Image: T7 promoter (strong promoter from T7 bacteriophage).jpg|center|frame|300px|<b>Figure 1. </b> Main principles of the molecular mechanism of T7 promoter]]<br><br>
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==<b>Design</b>==
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===Design Notes===
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We added some synonymous mutations to avoid part rules.
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===Source===
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We found this sequence data in the following renference.<br>
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*Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.<br><br>
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===References===
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Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.<br><br>
  
 
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Revision as of 04:53, 22 October 2020

xynD, which produces xylanase


T7 promoter is a very specific promoter which is transcribed only by specific T7 RNA polymerase which can activate downstream gene expression after being combined.

Usage and Biology

In our project, we divided T7 RNA polymerase into two parts, respectively called T7NT (链接) and T7CT (链接) and attached on the nMag and pMag. Under blue light irradiation, these two parts undergo specific blue light dependent heterodimerization and assemble into complete T7rna polymerase, which acts on PT7 promoter and activates downstream gene expression. And the process is reversible, that is, when blue light is stopped, the dimerized T7RNA polymerase de-dimers and loses its function.

The kanR gene is a relatively common type of resistance gene in molecular cloning, and it is also very common in commercial plasmids. In the process of constructing plasmids, we will choose kanamycin-resistant vectors, so we no longer too much detail on its mechanism of action.

Figure 1. Main principles of the molecular mechanism of T7 promoter



Design

Design Notes

We added some synonymous mutations to avoid part rules.


Source

We found this sequence data in the following renference.

  • Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.

References

Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 24
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]