DNA

Part:BBa_K4670134:Design

Designed by: Elon Mok Yat Lok   Group: iGEM23_HongKong-PuiChing   (2023-10-10)


Encodes a member of the GATA factor family of zinc finger transcription factors.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 197
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 197
    Illegal NheI site found at 283
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 197
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 197
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 197
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Keep the GC content lower than 35%

Eliminate the chance of forming stem loop

All exons are excluded

Removing all illegal restriction sites

Be careful of the design of the golden gate assembly recognition sites

Improvements made

We control the size of liposomes by Thermal treatment, and hence reduce the diameter of each liposome to under 10nm, eventually aid the entry of GFP fusion protein in to the liposome carrier. (refer to Results page for further analysis). Also, we sucessfully coded the plasmid design and run PCR and Golden Gate Assembly to validate our work. (refer to results page for proofs). In addition, the backbone we used before the standards of iGEM release is not optimal for our promotor to work on and hence reduce the expressing efficiency of GATA4 GFP composite gene, we then switched up to the pUC-AmpRv1 backbone as the bedrock of our fusion protein, facilitating the process.

engineering19.jpeg engineering17.jpg
Figure 6.a: original design Figure 6.b: enhanced design


Source

https://www.ncbi.nlm.nih.gov/gene/825224

References