Difference between revisions of "Part:BBa K2996503"
 
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<partinfo>BBa_K2996503 short</partinfo>
 
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This part is made up of part BBa_K2996502 and part BBa_K2996501,which produces a fused protein of C-luciferase and zincfinger.We used overlapping PCR to produce the DNA fragment.The fused protein is aimed to bring the C-luciferase to the site of target sequence of zincfinger.
 
This part is made up of part BBa_K2996502 and part BBa_K2996501,which produces a fused protein of C-luciferase and zincfinger.We used overlapping PCR to produce the DNA fragment.The fused protein is aimed to bring the C-luciferase to the site of target sequence of zincfinger.
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The product of this part will bind to the binding site of DNA binding domain,thus bringing the C-luciferase near the "lure" sequence in our project.The dCas9 fused with N-luciferase will bind to "lure" sequence once the off-target incidence takes place.When the two parts of  luciferase get near,they will make the whole enzyme.
  
 
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<center><img style="max-width: 80%" src="https://2019.igem.org/wiki/images/3/3d/T--SJTU-BioX-Shanghai--jsf-NCDBD.jpg" alt="" /></center>
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<center><b>Figure 1.</b> <i>overlap PCR product of C-luciferase-DBD(963bp). The ladder represents the size of DNA in bp.</i> </center>
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<center><b>Fig. 1 Overlap PCR product of C-luciferase and DBD.(Lane1:overlap PCR product;Lane 2:2000bp ladder)</i> </center>
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===Functional Parameters===
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<partinfo>BBa_K2996503 parameters</partinfo>
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<center><img style="max-width: 80%" src="https://2019.igem.org/wiki/images/8/89/T--SJTU-BioX-Shanghai--wet_lab-OT2.1.png" alt="" /></center>
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Fig. 2 The working pattern of BBa K2996503.

Latest revision as of 23:53, 21 October 2019


The part is aimed to create a fused protein of C-luciferase and zincfinger

This part is made up of part BBa_K2996502 and part BBa_K2996501,which produces a fused protein of C-luciferase and zincfinger.We used overlapping PCR to produce the DNA fragment.The fused protein is aimed to bring the C-luciferase to the site of target sequence of zincfinger. The product of this part will bind to the binding site of DNA binding domain,thus bringing the C-luciferase near the "lure" sequence in our project.The dCas9 fused with N-luciferase will bind to "lure" sequence once the off-target incidence takes place.When the two parts of luciferase get near,they will make the whole enzyme.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 846
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 302
    Illegal AgeI site found at 441
  • 1000
    COMPATIBLE WITH RFC[1000]


Fig. 1 Overlap PCR product of C-luciferase and DBD.(Lane1:overlap PCR product;Lane 2:2000bp ladder)</i> </center>


Fig. 2 The working pattern of BBa K2996503.