Device

Part:BBa_K4831015:Design

Designed by: Joakim Gustafsson   Group: iGEM23_ABOA-Turku   (2023-10-03)


IPTG inducible merA (Synechocystis) expression device with a EFE as a reporter gene


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1498
    Illegal SpeI site found at 1511
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1498
    Illegal SpeI site found at 1511
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1498
    Illegal BglII site found at 1612
    Illegal XhoI site found at 1481
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1498
    Illegal SpeI site found at 1511
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1498
    Illegal SpeI site found at 1511
    Illegal NgoMIV site found at 3788
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This device and all its parts are designed specifically for Synechocystis sp. PCC6803. That means that the codon presence is similar to Synechocystis native genes. This device is assembled with a golden gate-inspired assembly system, leaving specific 4 bps scars between every part.

Source

Multiple sites. Genomic parts of different organisms. All for Synechocystis sp. PCC6803

References

[1] Guerrero, F., Carbonell, V., Cossu, M., Correddu, D. and Jones. P. R. (2012) Ethylene synthesis and regulated expression of recombinant protein in Synechocystis sp. PCC 6803. PLoS ONE 7(11):e50470.http://dx.doi.org/10.1371/journal.pone.0050470

[2] Thiel, K., Mulaku, E., Dandapani, H., Nagy, C., Aro, E-M., Kallio, P. (2018) Translation efficiency of heterologous proteins is significantly affected by the genetic context of RBS sequences in engineered cyanobacterium Synechocystis sp. PCC 6803. Microbial Cell Factories 17(1):34. https://doi.org/10.1186/s12934-018-0882-2

[3] Thiel K, Patrikainen P, Nagy C, Fitzpatrick D, Pope N, Aro EM, Kallio P. (2019) Redirecting photosynthetic electron flux in the cyanobacterium Synechocystis sp. PCC 6803 by the deletion of flavodiiron protein Flv3. Microbial Cell Factories, 18: 189. https://doi.org/10.1186/s12934-019-1238-2

[4] Nagy, C., Thiel, K., Mulaku, E., Mustila, H., Tamagnini, P., Aro, E-M, Pacheco, C. C., Kallio, P. (2021) Comparison of alternative integration sites in the chromosome and the native plasmids of the cyanobacterium Synechocystis sp. PCC 6803 in respect to expression efficiency and copy number. Microbial Cell Factories 20, 130. https://doi.org/10.1186/s12934-021-01622-2

[5] Rehbein, P., Berz, J., Kreisel, P., & Schwalbe, H. (2019). "CodonWizard" - An intuitive software tool with graphical user interface for customizable codon optimization in protein expression efforts. Protein expression and purification, 160, 84–93. https://doi.org/10.1016/j.pep.2019.03.018