Part:BBa_K3765014

OpdA_Signal+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-sfGFP excluding the sfGFP module, which was removed by digestion with KpnI followed by ligation. As a result, the fusion protein encoded by this CDS is less bulky than its previous version.

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). This also implies that the SpyTag002 module is far away from the core enzyme and can reliably form an isopeptide bond with SpyCatcher/SpyCatcher002, and this binding is not expected to hamper the enzymatic activity of OpdA.

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

---

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 1219
  Illegal AgeI site found at 1282
  
• 1000
  COMPATIBLE WITH RFC[1000]