Part:BBa_K3699004

pylT(tRNA)

tRNA: the pylT gene (tRNA) of Methanosarcina mazei, proK promoter

Design

From virus to human, the same set of genetic code is employed, which contains the start codon, three stop codons (UAA, UAG, UGA), and the remaining 61 codons encoding 20 kinds of amino acids. However, some exceptions have been found, such as pyrrolysyl tRNA synthetase/tRNA (pylrs/tRNA) from Methanosarcina mazei, which can recognize the amber codon UAG and translate it into pyrrolysine. Moreover, in E. coli, yeasts, and mammalian cells, this PylRS/tRNA pair is orthogonal, which means it does not interfere with the translation system of the chassis [1]. The orthogonal translation system has been used in vaccine safety design [2]. Now, we will use it to achieve the anti-escape design of cyanophages.

References

[1] Mukai T , Kobayashi T , Hino N , et al. Adding l-lysine derivatives to the genetic code of mammalian cells with engineered pyrrolysyl-tRNA synthetases[J]. Biochemical & Biophysical Research Communications, 2008, 371(4):818-822.

[2] Si L , Xu H , Zhou X , et al. Generation of influenza A viruses as live but replication-incompetent virus vaccines.[J]. Science, 2016.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 230
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]