Coding

Part:BBa_K4249001:Design

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


The biosynthetic gene for ERG production from C. purpurea that catalyzing the 2nd enzymatic step


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1057
    Illegal XbaI site found at 403
    Illegal XbaI site found at 748
    Illegal XbaI site found at 1036
    Illegal PstI site found at 779
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1057
    Illegal PstI site found at 779
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1057
    Illegal BglII site found at 400
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1057
    Illegal XbaI site found at 403
    Illegal XbaI site found at 748
    Illegal XbaI site found at 1036
    Illegal PstI site found at 779
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1057
    Illegal XbaI site found at 403
    Illegal XbaI site found at 748
    Illegal XbaI site found at 1036
    Illegal PstI site found at 779
  • 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 encode egt2 that from ergot fungus Claviceps purpurea.

References