Part:BBa_K5142008

amber suppressor tRNA

Compared to leucine codons and tryptophan codons, the amber codon is one of the stop codons, and its main function is to send a termination signal during protein synthesis, instructing the ribosome to stop adding amino acids. This is a key mechanism to ensure that protein synthesis ends correctly.

Therefore, we use an amber suppressor tRNA, which reads the UAG stop codon, allowing the insertion of 4-L-azido-phenylalanine at this position. In this way, the UAG codon, which would normally cause the termination of protein synthesis, can now encode a non-natural amino acid. With the introduction of an artificial translation system and the addition of 4-L-azido-phenylalanine, we can synthesize a complete A27L protein with an azide group.

[1] Kensaku Sakamoto 1, Akiko Hayashi, Ayako Sakamoto, Daisuke Kiga, Hiroshi Nakayama, Akiko Soma, Takatsugu Kobayashi, Makoto Kitabatake, Koji Takio, Kazuki Saito, Mikako Shirouzu, Ichiro Hirao, Shigeyuki Yokoyama.Site-specific incorporation of an unnatural amino acid into proteins in mammalian cells[EB/OL].2002[2024-9-28].doi: 10.1093/nar/gkf589..

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 1137
  Illegal SpeI site found at 15
  Illegal PstI site found at 1556
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal prefix found in sequence at 1
  Illegal suffix found in sequence at 1543
  Illegal EcoRI site found at 1137
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 1137
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 1137
  Illegal SpeI site found at 15
  Illegal PstI site found at 1556
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 1137
  Illegal SpeI site found at 15
  Illegal PstI site found at 1556
  
• 1000
  COMPATIBLE WITH RFC[1000]