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:
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.