Part:BBa_K1408002

Gal4-Degron-Vp16

Gal4 protein binds to a Gal1 promoter while VP16 recruits transcription machinery promoting transcription to anything under the Gal1 promoter. The two must be co-localized to work as a transcriptional activator.

The degron protein domain when fused to a protein acts as a source of instability leading to degradation by the cell via ubiquitination.

This system can be used to compare the relative stabilities of proteins. A sequence for a protein inserted between the degron and VP16 will produce a fusion protein which acts as a transcriptional activator for anything under a Gal1 promoter. If the protein inserted is unstable it will be degraded by the cell and the Gal4-VP16 transcriptional activator will no longer function. If a quantifiable marker protein such as GFP is under Gal1 then the level of GFP output of the cell is related to the stability of the inserted protein.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 870
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 340
  Illegal XhoI site found at 85
  Illegal XhoI site found at 568
  Illegal XhoI site found at 1305
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 487