Device
Part:BBa_K4461505:Design
Designed by: SHIH-HSUN, YANG Group: iGEM22_NYCU-Taipei (2022-10-10)
glpABC+RBS+(mCherry+tag)+BBa_B0015
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 993
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 993
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 993
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 993
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 993
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We cloned the promoter from E.coli K-12 MG1655 by PCR. Then we used PCR again to add suffix and prefix. Also, we separated the plasmid that we want to insert the promoter into two parts. Therefore, we can use Gibson Assembly to ligate three fragments. Then, we mutated the stop codon on mCherry and added EcoRI and HindIII cutting site by PCR twice. At last, we can insert the commercial tag by enzyme digestion and ligation.
Source
gene synthesis