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