Generator
Part:BBa_K2239001:Design
Designed by: Ruohong Wang Group: iGEM17_SDSZ-China (2017-09-30)
ChBD-7alphaHSDH
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1041
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 875
Design Notes
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Source
The 7alpha-HSDH sequence is cloned from the genome of E.coli through PCR amplification, using the primers designed and synthesized based on its sequence. The restriction site BamH I is added to the upstream primer, and Hind III is added on the downstream primer. The CBD sequence is retrieved from the GeneBank, and is inserted into plasmid pUC57. The CBD gene is then cloned from the plasmid by PCR, with the restriction site Hind III added to the upstream primer, and Xhol I added to the downstream primer.