Part:BBa_K5005003
Expresses editor under dox induction.
AID-T7 RNAP-UGI is the key part of pEditor plasmid. It can express editor under doxycycline induction, which can edit genes downstream of the T7 promoter on the pTarget plasmids.
The directed evolution system TRACE was first reported by the Chen Fei research group at MIT. The key plasmid, pEditor (BBa_K5005003), used in our experiments, was provided by Professor Yu Yang from the Institute of Biophysics, Chinese Academy of Sciences. Firstly, we verified the feasibility of our directed evolution system (TRACE) using a customized EGFP plasmid. In principle, we mutated the 199th base of the EGFP gene from T-A to C-G, converting tyrosine to histidine and changing the fluorescence color from green to blue. Therefore, if the TRACE system is feasible, we can mutate the BFP gene obtained from site-directed mutagenesis back to the EGFP gene, achieving a change in cell fluorescence from blue to green.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1088
Illegal XhoI site found at 1822
Illegal XhoI site found at 2119 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2598
Illegal AgeI site found at 3322 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1308
Illegal BsaI site found at 2620
Illegal BsaI.rc site found at 76
Illegal BsaI.rc site found at 1094
Illegal BsaI.rc site found at 3319
Illegal SapI site found at 177
Illegal SapI.rc site found at 1129
Illegal SapI.rc site found at 1702
Illegal SapI.rc site found at 2727
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