Part:BBa_K4249003
The expression cassette carrying an TEF1 promoter, CpEgt2 coding sequence and an CYC1 terminator
This part is the gene expression cassettes corresponding to the above two genes respectively. The expression cassettes both carrying a promoter, gene coding sequence and a terminator. In order to enable the engineered strain to produce ergothioneine in a more sustainable way, we chose the constitutive promoter to introduce the expression of the two heterologous genes. According to literature references and existing resources in our laboratory, we finally chose apply pTEF1 and pADH1 to our work, they are commonly used for metabolic engineering of yeast, more importantly, both of them could provide relatively high expression values of genes, which means we could get more production of ergothioneine in the end. Transcription terminators play a role in terminating the progress of gene transcription, and are thus essential elements in the gene expression. For both gene expression cassettes, we chose apply tCYC to our work. In addition, it should be noted that we integrated these gene expression cassettes into the yeast genome via homology repair, which means the homology arm are required. Here, these two gene expression cassettes were added with ~0.5kb homology arms on both sides when they were used as the linear integration fragments.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1497
Illegal XbaI site found at 425
Illegal XbaI site found at 843
Illegal XbaI site found at 1188
Illegal XbaI site found at 1476
Illegal SpeI site found at 419
Illegal PstI site found at 1219
Illegal PstI site found at 2033 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1497
Illegal SpeI site found at 419
Illegal PstI site found at 1219
Illegal PstI site found at 2033 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1497
Illegal BglII site found at 840 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1497
Illegal XbaI site found at 425
Illegal XbaI site found at 843
Illegal XbaI site found at 1188
Illegal XbaI site found at 1476
Illegal SpeI site found at 419
Illegal PstI site found at 1219
Illegal PstI site found at 2033 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1497
Illegal XbaI site found at 425
Illegal XbaI site found at 843
Illegal XbaI site found at 1188
Illegal XbaI site found at 1476
Illegal SpeI site found at 419
Illegal PstI site found at 1219
Illegal PstI site found at 2033 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 168
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