Plasmid

Part:BBa_K5371202:Design

Designed by: Haoyang Yu   Group: iGEM24_iZJU-China   (2024-09-29)


GAL4WTV2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 4329
    Illegal XbaI site found at 1646
    Illegal PstI site found at 527
    Illegal PstI site found at 2289
    Illegal PstI site found at 4943
    Illegal PstI site found at 8417
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 4329
    Illegal PstI site found at 527
    Illegal PstI site found at 2289
    Illegal PstI site found at 4943
    Illegal PstI site found at 8417
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 4329
    Illegal BamHI site found at 48
    Illegal BamHI site found at 4060
    Illegal XhoI site found at 1158
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 4329
    Illegal XbaI site found at 1646
    Illegal PstI site found at 527
    Illegal PstI site found at 2289
    Illegal PstI site found at 4943
    Illegal PstI site found at 8417
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 4329
    Illegal XbaI site found at 1646
    Illegal PstI site found at 527
    Illegal PstI site found at 2289
    Illegal PstI site found at 4943
    Illegal PstI site found at 8417
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1077
    Illegal BsaI.rc site found at 2753
    Illegal BsaI.rc site found at 3942
    Illegal BsaI.rc site found at 6668
    Illegal BsaI.rc site found at 7727
    Illegal SapI site found at 1442
    Illegal SapI site found at 5585
    Illegal SapI site found at 7879


Design Notes

Stop codons of Sir2 homologous region and mCherry were removed for continuous and complete expression of Sir2-linker-mcherry to prevent a premature stop in this case. Length of the linker is adjusted to avoid frameshift, which may cause faulty translation. Codons of the linker is designed to avoid polarized amino acids or those with large residues.


Source

GAL4WTV2 was constructed via Gibson Assembly, with segments mainly amplified from Saccharomyces cerevisiae.

References