Terminator

Part:BBa_K4451002:Design

Designed by: Brooks J Rady   Group: iGEM22_Sheffield   (2022-09-30)


tZ (T3 + rrnBT1 + T7TΦ UUCG)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 112
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Two nucleotide edits were made by Team Sheffield 2022 to the nucleotide sequence of the T7TΦ UUCG stem loop to allow RFC1000 compatibility. Regretfully, we did not have the opportunity to demonstrate terminator function before the project deadline. This variant was synthesised as dsDNA by Integrated DNA Technologies (with some difficulty: due to the complex secondary structure and repetitive nature of tZ, the addition of flanking adapter sequences was required to pass IDT quality control).

Source

https://doi.org/10.1021/sb5000115

References

Mairhofer, J., Wittwer, A., Cserjan-Puschmann, M. & Striedner, G. Preventing T7 RNA Polymerase Read-through Transcription—A Synthetic Termination Signal Capable of Improving Bioprocess Stability. ACS Synth. Biol. 4, 265–273 (2015).