Plasmid
ADH1p-DmNa

Part:BBa_K4203002:Design

Designed by: Yixuan Wang ;Zongjun Lee   Group: iGEM22_BJWZ-China   (2022-10-12)


After codon optimization, those part is suitable for the fusion protein ADH1p-DmNan-P2A-mCherry expr


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3181
    Illegal XbaI site found at 3893
    Illegal XbaI site found at 4886
    Illegal XbaI site found at 9224
    Illegal PstI site found at 1183
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3181
    Illegal NheI site found at 1008
    Illegal NheI site found at 6053
    Illegal PstI site found at 1183
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3181
    Illegal BglII site found at 899
    Illegal BglII site found at 959
    Illegal BglII site found at 3320
    Illegal BamHI site found at 5961
    Illegal XhoI site found at 6031
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3181
    Illegal XbaI site found at 3893
    Illegal XbaI site found at 4886
    Illegal XbaI site found at 9224
    Illegal PstI site found at 1183
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3181
    Illegal XbaI site found at 3893
    Illegal XbaI site found at 4886
    Illegal XbaI site found at 9224
    Illegal PstI site found at 1183
    Illegal NgoMIV site found at 1748
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4269
    Illegal BsaI.rc site found at 7394
    Illegal SapI site found at 3252
    Illegal SapI site found at 6311


Design Notes

Pay attention to the selection of Linker length and species, otherwise the efficiency of protein expression will be seriously affected. Noted that after expression the P2A would cut the mCherry off.


Source

The sequence of ADH1 promoter comes from pAdh(Strong Expression level Constitutive promoter in yeast) The sequence of DmNan component comes from Drosophila melanogaster (Ensembl:CG5842) , A TRPV family ion channel required for hearing in Drosophila. The sequence of P2A comes from pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP. The sequence of mCherry comes from POM42-mCherry.


References