Part:BBa_K511000

Hef1a-LacOid Promoter MammoBlock

This part encodes a promoter with low-level, constitutive activity that can be repressed by variants of the LacI transcriptional repressor. This part is a significant improvement over the existing Part:BBa_K415508, as it adds repressibility to the existing well-known constitutive promoter. In addition, different domains can be added onto the LacI repressor or IPTG can be added, allowing the user to toggle and tune the amount of repression, significantly increasing the flexibility and usage of the existing part.

Cotransfection of a construct containing a yellow fluorescent protein (EYFP) driven by the Hef1a-LacO promoter with a construct containing a LacI-KRAB transcriptional repressor driven by the TRE-Tight promoter proves that it is possible to repress Hef1a-LacO expression with the LacI-KRAB LacI variant, which includes a Kruppel-associated box motif previously shown to silence chromatin and thereby inhibit transcription.

To see further information about this part, consult the Results found on the iGEM 2011 wiki.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 416
  Illegal PstI site found at 921
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 292
  Illegal PstI site found at 416
  Illegal PstI site found at 921
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 670
  Illegal BamHI site found at 255
  Illegal XhoI site found at 1069
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 416
  Illegal PstI site found at 921
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 416
  Illegal PstI site found at 921
  Illegal NgoMIV site found at 804
  Illegal AgeI site found at 80
  
• 1000
  COMPATIBLE WITH RFC[1000]