Part:BBa_K4165201

Trim-(GSGS)-Coh2

This parts code for the trim21 E3 ligase having his PRYSPRY domain truncated (BBa_K3396007), fused to type 1 Cohesin module derived from Clostridium thermocellum cellulosome scaffoldin using Glycine serine flexible linker repeated two times to maintain part flexibility needed during target ubiquitination

Source

Synthetic Fusion protein

Sequence and Features

the sequence was optimized for E.coli expression

Assembly Compatibility:

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

  Illegal NheI site found at 225
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 182
  
• 1000
  COMPATIBLE WITH RFC[1000]

Dry Lab

Modeling

tTrim21-GSGS-Coh2 is modeled by AlphaFold2, ITASSER, Robetta and TrRosetta, best model obtained from TrRosetta.

                            Figure 1.: Predicted 3D structure of our fusion protein tTrim21-GSGS-Coh2.

Table 1: Quality assessment parameters of tTrim21-GSGS-Coh2. model.

References

1- Carvalho AL, Dias FM, Nagy T, Prates JA, Proctor MR, Smith N, Bayer EA, Davies GJ, Ferreira LM, Romão MJ, Fontes CM, Gilbert HJ. Evidence for a dual binding mode of dockerin modules to cohesins. Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3089-94. doi: 10.1073/pnas.0611173104. Epub 2007 Feb 20. PMID: 17360613; PMCID: PMC1805526.