Difference between revisions of "Part:BBa K3365001"
 
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<partinfo>BBa_K3365001 short</partinfo>
 
<partinfo>BBa_K3365001 short</partinfo>
  
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This part is used to detect whether dCas9 protein is on-target.The sequence is made up of <i>PDCD1</i>, which is the binding site of dCas9, and J23117 BBa_J23117 (visit: [https://parts.igem.org/Part:BBa_J23117]) weak promoter spaced by 35 base pairs. The dCas9 in part BBa_K3365001 (visit: [https://parts.igem.org/Part:BBa_K3365001]) could bind to the <i>PDCD1</i>, wihch as the target of dCas9,  under guidence of specific designed sgRNA, while subunit Omega could bind to J23117 and activating the expression of downsream fluorescent protein. According to reference, we knew that 35 base pairs is the best spacer for activating, so we chose 35 base pairs in plasmid pUC57 at random.
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This part is used to detect whether dCas9 protein is on-target. The sequence is made up of <i>PDCD1</i>, which is the binding site of dCas9, and J23117  
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<a href="https://parts.igem.org/Part:BBa_J23117">BBa_J23117</a>
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</html>
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<br/>
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weak promoter spaced by 35 base pairs. The dCas9 in this part could bind to the <i>PDCD1</i>, under guidance of specific designed sgRNA, while subunit Omega could bind to J23117 and activating the expression of downstream fluorescent protein. According to reference, we knew that 35 base pairs is the best length of spacer for activating, so we randomly choose 35bp sequence from the plasmid pUC57 as spacer.
  
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[[File:Target_binding_site_for_dCas9_and_subnit_Omega.png]]
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===Usage and Biology===
  
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It is the target binding site for dCas9 fused with subnit Omega in transcription acvating system.
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<figcaption><b>Fig.1:</b> Schematic diagram of this part
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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Latest revision as of 17:48, 27 October 2020


Target binding site for dCas9 and subnit Omega

This part is used to detect whether dCas9 protein is on-target. The sequence is made up of PDCD1, which is the binding site of dCas9, and J23117 BBa_J23117
weak promoter spaced by 35 base pairs. The dCas9 in this part could bind to the PDCD1, under guidance of specific designed sgRNA, while subunit Omega could bind to J23117 and activating the expression of downstream fluorescent protein. According to reference, we knew that 35 base pairs is the best length of spacer for activating, so we randomly choose 35bp sequence from the plasmid pUC57 as spacer.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 66
    Illegal NheI site found at 89
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]