Part:BBa_K5416062

pT7LacO + P22 + HRT2-SP + T7term

Imperial-College 2024

This part is designed by Team Imperial_College in iGEM 2024. It is reported to form an artificial organelle which serves as an enclosed chamber for natrual rubber production.

This is a composite part consisting of two CDS: P22 and HRT2-SP protein, placed downstream of pT7-LacO promoter (BBa_K2406020) for inducible overexpression. P22-His (BBa_K5416061) is the bacteriophage P22 mature virion capsid protein fused with a His-tag. The HRT2-SP encodes a truncated cis-prentyltransferase HRT2trunc(BBa_K5416000) derived from H. brasiliensis that is recombined with a scaffold-protein (SP, BBa_K3187017) domain that can interact with the inner surface of the P22 capsid. We here to report this designed composite part to be capable of forming a virus-like-particle (VLP) as an artificial organelle with a compartmentalized environment for rubber production.

We have shown this part to have the following functions:

• Forming purifiable artifical organelle
• Encapsulating enzymes
• Producing hydrophibc internal environment
• Producing small amount of rubber

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 NgoMIV site found at 636
  Illegal AgeI site found at 1037
  
• 1000
  COMPATIBLE WITH RFC[1000]