Coding

Part:BBa_K5063014:Design

Designed by: Wong Chung Kiu   Group: iGEM24_HK-United   (2024-09-30)


GFP_TP1_P5_insert_sequence


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 741


Design Notes

T7 promoter can be recognized by T7 RNA polymerase, a very active enzyme. The T7 RNA Polymerase/Promoter System provides a convenient method for expression in a recombinant protein. The T7 promoter is commonly used to regulate gene expression of recombinant proteins, which can be subsequently used for a variety of downstream research applications. His-tag is chosen as the affinity tag for our fusion protein due to its small size. This prevents the function or structure of our protein from being interfered. His-tags also provide inexpensive methods to perform purification. The green fluorescent protein (GFP), is a secondary fluorescent protein which produces green light while doing an energy transfer reaction. GFP is widely used as a marker of gene expression and protein localization in living and fixed tissues due to its intense fluorescence.



Source

[1] Leong, D. P., Joseph, P. G., McKee, M., Anand, S. S., Teo, K. K., Schwalm, J. D., & Yusuf, S. (2017). Reducing the global burden of cardiovascular disease, part 2: prevention and treatment of cardiovascular disease. Circulation research, 121(6), 695-710.https://www.ahajournals.org/doi/epub/10.1161/CIRCRESAHA.117.311849 [2] Loo, S., Kam, A., Dutta, B., Zhang, X., Feng, N., Sze, S. K., ... & Tam, J. P. (2024). Broad-spectrum ginsentides are principal bioactives in unraveling the cure-all effects of ginseng. Acta Pharmaceutica Sinica B, 14(2), 653-666.https://www.sciencedirect.com/science/article/pii/S2211383523004227 [3] Dutta B, Loo S, Kam A, Sze SK, Tam JP. Ginsentide TP1 Protects Hypoxia-Induced Dysfunction and ER Stress-Linked Apoptosis. Cells. 2023 May 16;12(10):1401. doi: 10.3390/cells12101401. PMID: 37408235; PMCID: PMC10216702. [4] Lammi C, Fassi EMA, Li J, Bartolomei M, Benigno G, Roda G, Arnoldi A, Grazioso G. Computational Design and Biological Evaluation of Analogs of Lupin Peptide P5 Endowed with Dual PCSK9/HMG-CoAR Inhibiting Activity. Pharmaceutics. 2022 Mar 18;14(3):665. doi: 10.3390/pharmaceutics14030665. PMID: 35336039; PMCID: PMC8951016.

References