Difference between revisions of "Part:BBa K5142033"

 
 
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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 amber suppressor tRNA gene of Bacillus stearothermophilus.</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'>To provide sufficient tRNAs for mutant A27L synthesis, we input nine copies of the amber suppressor tRNA gene tandemly in this part. Each amber suppressor tRNA gene contain an RNA polymerase III promoter, a DNA sequence of amber suppressor tRNA and an RNA polymerase III terminator.</span>
 
<partinfo>BBa_K5142033 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5142033 SequenceAndFeatures</partinfo>
  

Latest revision as of 11:55, 2 October 2024


Amber suppressor tRNA gene tandem

This part contains nine identical genes which can be transcribed into amber suppressor tRNA with an anticondon of CUA so that it can read the amber suppressor codon UAG, allowing us to introduce 4-azido-L-phenylalanine to the mutant A27L in this project.

This part was subcloned from the plasmid pcpAzpaRS_v2 (Addgene Plasmid #196485) (1), which was originally derived from the amber suppressor tRNA gene of Bacillus stearothermophilus.
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.

To provide sufficient tRNAs for mutant A27L synthesis, we input nine copies of the amber suppressor tRNA gene tandemly in this part. Each amber suppressor tRNA gene contain an RNA polymerase III promoter, a DNA sequence of amber suppressor tRNA and an RNA polymerase III terminator.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 1545
    Illegal EcoRI site found at 1139
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1139
    Illegal SpeI site found at 1546
    Illegal PstI site found at 1560
    Illegal NotI site found at 7
    Illegal NotI site found at 1553
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1139
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 1546
    Illegal EcoRI site found at 1139
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1
    Illegal EcoRI site found at 1139
    Illegal XbaI site found at 16
    Illegal SpeI site found at 1546
    Illegal PstI site found at 1560
  • 1000
    COMPATIBLE WITH RFC[1000]