Coding

Part:BBa_K4249000:Design

Designed by: Douglas B.Kell and Irina Borodina   Group: iGEM22_HBUT-China   (2022-10-11)


The biosynthetic gene for ERG production from N.crassa that catalyzing the first enzymatic step


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 751
    Illegal EcoRI site found at 2078
    Illegal XbaI site found at 1435
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 751
    Illegal EcoRI site found at 2078
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 751
    Illegal EcoRI site found at 2078
    Illegal BglII site found at 613
    Illegal BglII site found at 1099
    Illegal BglII site found at 1432
    Illegal BglII site found at 2101
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 751
    Illegal EcoRI site found at 2078
    Illegal XbaI site found at 1435
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 751
    Illegal EcoRI site found at 2078
    Illegal XbaI site found at 1435
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

During the experiment, we found that some regions of the promoter sequence templates are rich in GC residues, which tend to fold into complex secondary structures and might not melt during the annealing phase of the PCR cycle. Also, the primers used to amplify GC-rich regions often have a high capacity to form self- and cross-dimers and a strong tendency to fold into stem-loop structures that can impede the progress of the DNA polymerase along the template molecule. Therefore, instead of PCR method, we obtained the gene expression cassettes by enzyme digestion.


Source

The egt1 is gene coding sequence, that from Neurospora crassa, a filamentous fungus that produces ergothioneine.

References