Difference between revisions of "Part:BBa K2996008"
 
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<partinfo>BBa_K2996008 short</partinfo>
 
<partinfo>BBa_K2996008 short</partinfo>
  
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The expresssion of cas1-cas2 complex can be triggered by the addition of AHL.
 
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#Usage and Biology
 
 
Expression of the CRISPR adaptation proteins Cas1 and Cas2 promotes unidirectional integration of 33–base pair DNA spacers into our designed RSRL array.
 
Expression of the CRISPR adaptation proteins Cas1 and Cas2 promotes unidirectional integration of 33–base pair DNA spacers into our designed RSRL array.
  
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On the other hand, the input information is now acyl-homoserine lactones. In the transcription activation gene circuit, luxI can be put downstream of promotor J23001, enabling the expression of Luxl enzyme when dcas9 binds to lure sequence. AHL molecule production will then activate the expression of cas1-cas2 complex, in this way, the off-target incidence can be recorded.
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Our pLuxR-cas plasmid has pLuxR upstream of cas1-cas2, addition of AHL (acyl-homoserine lactones) will activate the expression of cas1-cas2 complex, thus accomplishing information storage.
  
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So, the input information is now acyl-homoserine lactones. In the transcription activation gene circuit, LuxI can be put downstream of promotor J23117, enabling the expression of Luxl enzyme when dcas9 binds to lure sequence. AHL molecule production will then activate the expression of cas1-cas2 complex, in this way, the off-target incidence can be recorded.
  
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#Result
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===Result===
 
蛋白胶图
 
蛋白胶图
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<html>
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<img style="max-width: 20%" src="https://static.igem.org/mediawiki/parts/8/89/NISIN3_4.png" alt="">
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<center><img style="max-width: 50%" src="https://static.igem.org/mediawiki/parts/8/89/NISIN3_4.png" alt="" /></center>
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</html>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 05:12, 20 October 2019


pLuxR upstream of cas1/2

Expression of the CRISPR adaptation proteins Cas1 and Cas2 promotes unidirectional integration of 33–base pair DNA spacers into our designed RSRL array.

Our pLuxR-cas plasmid has pLuxR upstream of cas1-cas2, addition of AHL (acyl-homoserine lactones) will activate the expression of cas1-cas2 complex, thus accomplishing information storage.

So, the input information is now acyl-homoserine lactones. In the transcription activation gene circuit, LuxI can be put downstream of promotor J23117, enabling the expression of Luxl enzyme when dcas9 binds to lure sequence. AHL molecule production will then activate the expression of cas1-cas2 complex, in this way, the off-target incidence can be recorded.


Result

蛋白胶图

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 258
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 979
  • 1000
    COMPATIBLE WITH RFC[1000]