Part:BBa_K5142033
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.
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 1545
Illegal EcoRI site found at 1139 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
Illegal EcoRI site found at 1139 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 1546
Illegal EcoRI site found at 1139 - 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]
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