T7

Part:BBa_K3192012:Design

Designed by: Simonne Guenette   Group: iGEM19_Virginia   (2019-10-12)


T7A1 Promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part DNA sequence was taken from NCBI and codon optimized for E. coli K12 chassis. The sequence was then reviewed for any illegal restriction sites and conservative edits were made as necessary. The coding sequence was then designed for Golden Gate (using Teselagen Software) assembly and then synthesized. This part was assembled into a larger composite part using Golden Gate Assembly and then sequenced for identity confirmation.

This promoter is intended for inducible expression of a coding region. If your desired gene(s) of expression need to be expressed constitutively do not use this promoter.

This promoter is ideal for high expression at specified intervals, as the addition of IPTG results in high levels of transcription. If metabolic load on your chassis is a concern for you team, this promoter is ideal for controlling the energy used on BrioBrick expression in a chassis.

Source

Escherichia coli

References

Sclavi, B. et al. Real-time characterization of intermediates in the pathway to open complex formation by Escherichia coli RNA polymerase at the T7A1 promoter. Proceedings of the National Academy of Sciences 13, 4706–4711 (2005).