Coding

Part:BBa_K5218002:Design

Designed by: Amy Fu, Juliette, Lichuan Chen, Xiaojuan Wang   Group: iGEM24_BGI-MammothEdu-South   (2024-08-11)


AtC3H


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 234
    Illegal NheI site found at 552
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 406
    Illegal XhoI site found at 749
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 132
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1193
    Illegal SapI site found at 387


Design Notes

The cloning strategy for AtC3H-GFP fusion gene is as follows: two rounds of primers pairs were used. The first reverse primer is designed on the 3' UTR of AtC3H CDS for specificity. The second round PCR uses first round PCR product as template, and the second reverse primer is located before the AtC3H CDS stop codon, and recombination overhang is added for cloning.

Source

The AtC3H gene ID is AT2G40890, transcript ID is NM_180006.2 (NCBI). Team BGI-MammothEdu-South 2024 extracted Arabidopsis thaliana total RNA, reverse transcribed into cDNA and cloned the AtC3H CDS with specific primer pairs.

References