Part:BBa_K1051278

The measurement of PEST of cln1.

Purpose

The measurement of PEST of cln1.

Principle

PEST,Certain amino acid sequences appear to be signals for degradation. One such sequence is known as the PEST sequence because short stretch of about eight amino acids is enriched with proline(P), glutamic acid(E), serine(S), and threonine(T) at C-terminal of proteins that are more rapidly degraded than other proteins.

Engineering

We convert base "A" to "G" at 348 site of cln1-PEST(K1051202) by overlap PCR.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 150
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1199
  

References

[1] Li X, Zhao X, Fang Y, Jiang X, Duong T, Fan C, Huang CC, Kain SR(1998) Generation of destabilized green fluorescent protein as a transcription reporter. J Biol Chem 273:34970–34975.
[2] Dorota Skowyra, Karen L Craig, Mike Tyers, Stephen J Elledge, J.Wade Harper "F-Box Proteins Are Receptors that Recruit Phosphorylated Substrates to the SCF Ubiquitin-Ligase Complex".