Composite
Part:BBa_K2624005:Design
Designed by: Shuyao Zhou, Huandi Xu, Junqing Zhang, Yali Wu Group: iGEM18_CPU_CHINA (2018-08-17)
hulc-pri-miRNA(MAP4K4)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 130
Design Notes
We checked the exact transcription start site of the hulc gene and replaced the sequence after it with the sequence that we designed so that, instead of the lncRNA, it would express the pri-miRNA analogue. Then of course we added a SV40 poly(A) sequence that signals the end of a transcriptional unit. In the end we added 5 different restriction sites for 5 different endonucleases, three in the front and two at the end, for recombination convenience.
Source
Hulc promoter sequence comes from Swissregulon Database. The pri-miRNA analogue part is designed by us.