Plasmid
TDH3p-DmIa

Part:BBa_K4203001: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 TDH3p-DmIav-sunTag-BFP expre


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 5086
    Illegal XbaI site found at 12102
    Illegal PstI site found at 5325
    Illegal PstI site found at 6711
    Illegal PstI site found at 6717
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 5086
    Illegal NheI site found at 8931
    Illegal PstI site found at 5325
    Illegal PstI site found at 6711
    Illegal PstI site found at 6717
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 5086
    Illegal BglII site found at 6449
    Illegal BglII site found at 7280
    Illegal BglII site found at 7295
    Illegal XhoI site found at 8909
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 5086
    Illegal XbaI site found at 12102
    Illegal PstI site found at 5325
    Illegal PstI site found at 6711
    Illegal PstI site found at 6717
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 5086
    Illegal XbaI site found at 12102
    Illegal PstI site found at 5325
    Illegal PstI site found at 6711
    Illegal PstI site found at 6717
    Illegal NgoMIV site found at 2799
    Illegal AgeI site found at 1503
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5266
    Illegal BsaI.rc site found at 10272
    Illegal SapI site found at 9189
    Illegal SapI.rc site found at 7209


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 TagBFP off.


Source

Saccharomyces cerevisiae S288C glyceraldehyde-3-phosphate dehydrogenase (phosphorylating) TDH3 (TDH3), partial mRNA(NM_001181321.3) The sequences of sunTag, TagBFP and P2A come from pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP. DmIav component comes from Drosophila melanogaster inactive (iav), partial mRNA(NM_132125.2)

References