Coding
Part:BBa_K4343118
Designed by: Xiaolu Sun Group: iGEM22_NNU-China (2022-10-09)
hisG-URA-hisG
HisG tag were utilized in combination with URA3 gene to recycle this screening marker for further use in the same strain. To be more specific, 5-fluoroorotic acid (5-FOA) could be converted into a toxic form (e.g., 5-fluorouracil) by functional OMP decarboxylase encoded by URA3 gene, thus making yeast cells unable to grow. To survive the toxicity, yeast would miss the URA3 gene by HigG tag through negative screening. In this circumstance, URA deficient strain allows for the next transformation.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1920
Illegal XhoI site found at 1953 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2108
Illegal AgeI site found at 97
Illegal AgeI site found at 458
Illegal AgeI site found at 3161
Illegal AgeI site found at 3522 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1
Illegal BsaI site found at 638
Illegal BsaI site found at 3702
Illegal SapI.rc site found at 363
Illegal SapI.rc site found at 3427
[edit]
Categories
Parameters
None |