Difference between revisions of "Part:BBa K5142034"

 
 
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>This part was subcloned from the plasmid pcpAzpaRS_v2 (Addgene Plasmid #196485) (1), which was originally derived from the TyrRS gene of E.coli.</span>
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<span class='h3bb'>Reference</span>
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<span class='h3bb'>1. Hino N, Sakamoto K, Yokoyama S. Site-specific incorporation of unnatural amino acids into proteins in mammalian cells. Methods Mol Biol. 2012;794:215-28. </span>
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<span class='h3bb'>pAzpaRS was designed to specifically aminoacylate the amber suppressor tRNA (see BBa_K5142008) without cross-reacting with endogenous tRNAs.</span>
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<partinfo>BBa_K5142034 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5142034 SequenceAndFeatures</partinfo>
  

Latest revision as of 11:57, 2 October 2024


Azidophenylalanine-specific tyrosyl-tRNA synthetase

This part includes the coding sequence (cds) of an azidophenylalanine-specific variant of E. coli tyrosyl-tRNA synthetase (pAzpaRS). pAzpaRS can transfer 4-azido-L-phenylalanine (4AzF) to the amber suppressor tRNA, forming the corresponding aminoacyl-tRNA. We used pAzpaRS to synthesize the amber suppressor tRNA carrying 4AzF, which was used to translate the amber suppressor codons in A27L-3stop gene, thereby inserting 4AzF into the mutant A27L protein.

This part was subcloned from the plasmid pcpAzpaRS_v2 (Addgene Plasmid #196485) (1), which was originally derived from the TyrRS gene of E.coli.
Reference

1. Hino N, Sakamoto K, Yokoyama S. Site-specific incorporation of unnatural amino acids into proteins in mammalian cells. Methods Mol Biol. 2012;794:215-28.

pAzpaRS was designed to specifically aminoacylate the amber suppressor tRNA (see BBa_K5142008) without cross-reacting with endogenous tRNAs.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 1293
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal SpeI site found at 1294
    Illegal PstI site found at 1308
    Illegal NotI site found at 7
    Illegal NotI site found at 1301
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal BamHI site found at 758
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 1294
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1
    Illegal XbaI site found at 16
    Illegal SpeI site found at 1294
    Illegal PstI site found at 1308
  • 1000
    COMPATIBLE WITH RFC[1000]