Device

Part:BBa_K4628041:Design

Designed by: Linghao Xia   Group: iGEM23_ZJU-China   (2023-10-11)


pGreenⅡ 0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 258
    Illegal EcoRI site found at 2663
    Illegal EcoRI site found at 4242
    Illegal XbaI site found at 987
    Illegal XbaI site found at 3347
    Illegal SpeI site found at 1206
    Illegal SpeI site found at 3209
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 258
    Illegal EcoRI site found at 2663
    Illegal EcoRI site found at 4242
    Illegal NheI site found at 1910
    Illegal SpeI site found at 1206
    Illegal SpeI site found at 3209
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 258
    Illegal EcoRI site found at 2663
    Illegal EcoRI site found at 4242
    Illegal BglII site found at 2609
    Illegal BglII site found at 3344
    Illegal BglII site found at 4172
    Illegal BglII site found at 4267
    Illegal BglII site found at 4978
    Illegal BamHI site found at 3320
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 258
    Illegal EcoRI site found at 2663
    Illegal EcoRI site found at 4242
    Illegal XbaI site found at 987
    Illegal XbaI site found at 3347
    Illegal SpeI site found at 1206
    Illegal SpeI site found at 3209
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 258
    Illegal EcoRI site found at 2663
    Illegal EcoRI site found at 4242
    Illegal XbaI site found at 987
    Illegal XbaI site found at 3347
    Illegal SpeI site found at 1206
    Illegal SpeI site found at 3209
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4726


Design Notes

By mutating the start codon of the capsid protein in the subPromoter CP sequence, we avoid the expression of some non-sense peptide chains of CP.


Source

The cloned TMV RNA dependent RNA polymerase-movement protein-subPromoter CP and eGFP were homologous recombined. Homologous recombination fragments are inserted in pGreen II backbone. Finally, the start codon in subPromoter CP was mutated by PCR and we get the target Part.

References