RNA

Part:BBa_K5142033:Design

Designed by: Yuchen Liang   Group: iGEM24_HunanU   (2024-09-30)


Amber suppressor tRNA gene tandem


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]


Design Notes

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.


Source

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.

References