Part:BBa_K2996503

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.