Part:BBa_K3765015

OpdA+22aaL+SpyTag002+10xHis

CDS of fusion protein consisting of SpyTag002 linked to OpdA (phosphotriesterase), which can covalently bind to SpyCatcher/SpyCatcher002. In our project, the protein encoded by this part is designed to get anchored to a SpyCatcher002-dCBD fusion protein.

Usage and Biology

This CDS part is the same as OpdA-SpyTag002 excluding the OpdA signal peptide. It was generated by amplification of the rest of the CDS of the previous OpdA-SPyTag002 CDS by PCR.

Fig 1. Feature Map

A tertiary structure for this fusion protein was predicted using AlphaFold. The OpdA module folds as in its native state, while the other stretches of amino acids exist as intrinsically disordered regions (IDRs). The SpyTag002/SpyCatcher002 binding is unaffected. Due to the absence of the considerably hydrophobic OpdA signal peptide which was predicted to exist as an intrinsically disordered region (IDR) by AlphaFold previously, the fusion protein encoded by this CDS is expected to be more stable and soluble in physiological conditions.

Fig 2. Predicted cartoon model [N- to C- direction is blue to red]

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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 1138
  Illegal AgeI site found at 1201
  
• 1000
  COMPATIBLE WITH RFC[1000]