Composite

Part:BBa_K5487004:Design

Designed by: Haibin Wu   Group: iGEM24_UESTC-China   (2024-09-01)


T7 Promoter+lac operator+RBS+TphA2+RBS+TphA1+Term+T7 promoter+lac operator+RBS+TphA3+RBS+TphB2+Term


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2326
    Illegal NheI site found at 2391
    Illegal NheI site found at 4055
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1457
    Illegal NgoMIV site found at 1766
    Illegal NgoMIV site found at 3291
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 3163


Design Notes

Term (BBa_1719005) was chosen because it is a terminator, which can effectively terminate transcription and protect mRNA from degradation. Different restriction sites were added between each part to facilitate the connection of each part or further manipulation. RBS (BBa_K587117) was chosen because it has a high translation initiation rate and is compatible with most bacterial hosts.Lac operator was chosen because it can regulate the expression of downstream genes according to the presence or absence of lactose, thus achieving inducible control.

Source

Lac Operator: From the lac operon of Escherichia coli.RBS: From the standard biobrick provided by NEB company.TphA1, TphA2, TphA3, and TphB2 : From the genomic sequence of Comamonas sp.Term: From the standard biobrick provided by NEB company.

References

Tramontina, R.; Galman, J. L.; Parmeggiani, F.; Derrington, S. R.; Bugg, T. D. H.; Turner, N. J.; Squina, F. M.; Dixon, N., Consolidated production of coniferol and other high-value aromatic alcohols directly from lignocellulosic biomass. Green Chemistry 2020, 22 (1), 144-152.