DNA

Part:BBa_K4610006:Design

Designed by: Sophie Geierstanger   Group: iGEM23_CCA-San-Diego   (2023-10-07)


Afu8g00380 (Fma-AT)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 882
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 882
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 169
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 882
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 882
    Illegal NgoMIV site found at 81
    Illegal NgoMIV site found at 281
    Illegal NgoMIV site found at 767
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 408


Design Notes

In order to increase the rate of transfection, our team has decided to construct two 4 layer chitosan nanocomposites that contain in total 8 DNA sequences rather than one 8 DNA layer nanocomposite. The first chitosan nanocomposite, chitosan nanocomposite A, would contain parts BBa_K4610000, BBa_K4610001, BBa_K4610002, and BBa_K4610003 (in order from outer to inner layer). The second chitosan nanocomposite, chitosan nanocomposite B, would contain BBa_K4610004, BBa_K4610005, BBa_K4610006, and BBa_K4610007 (in order from outer to inner layer).

In the constructed chitosan nanocomposite B, BBa_K4610006 (Afu8g00380 encoding for Fma-AT) is the third innermost layer. After its degradation and release, the core layer, BBa_K4610007 will be released. BBa_K4610006 is released after the sixth layer, BBa_K4610005 (Fma-PKS) is released into the cell.

Source

The source of Afu8g00380 is the aspergillus fumigatus strain Af293, a saprophytic filamentous fungus. Afu8g00380 is located in chromosome 8. Af293 is a clinically derived strain that was isolated in 1994 from a lung biopsy taken post autopsy from a patient who was developing severe sepsis syndrome. In 1998 it became the first A. fumigatus isolate with a full genome sequence and even in 202, Af293 genome is the only genome sequence of A. fumigatus to have a full physical reconstruction of its chromosomes. Af293 has eight chromosomes, however, Afu8g00380 exists on the 8th chromosome.

References

Guruceaga, X., Perez-Cuesta, U., Abad-Diaz de Cerio, A., Gonzalez, O., Alonso, R. M., Hernando, F. L., Ramirez-Garcia, A., & Rementeria, A. (2019, December 20). Fumagillin, a mycotoxin of aspergillus fumigatus: Biosynthesis, biological activities, detection, and applications. Toxins. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7020470/

Bertuzzi, M., van Rhijn, N., Krappmann, S., Bowyer, P., Bromley, M. J., & Bignell, E. M. (2020, September 17). On the lineage of aspergillus fumigatus isolates in common laboratory use. OUP Academic. https://academic.oup.com/mmy/article/59/1/7/5908083