Difference between revisions of "Part:BBa K3140003:Design"
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===Source=== | ===Source=== | ||
| − | The native sequence for the | + | The native sequence for the norbaeocystin methyltransferase PsiM (''Psilocybe cubensis'') was obtained from NCBI: GenBank accession [https://www.ncbi.nlm.nih.gov/nuccore/KY984100.1 KY984100.1] |
===References=== | ===References=== | ||
'''1.''' Fricke, J., Blei, F. & Hoffmeister, D. Enzymatic Synthesis of Psilocybin. ''Angew Chem Int Ed Engl'' '''56''', 12352-12355 (2017). | '''1.''' Fricke, J., Blei, F. & Hoffmeister, D. Enzymatic Synthesis of Psilocybin. ''Angew Chem Int Ed Engl'' '''56''', 12352-12355 (2017). | ||
Latest revision as of 06:08, 15 October 2019
PsiM - Norbaeocystin methyltransferase from Psilocybe cubensis
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part was intended for use in pET-28c(+) for initial analysis of protein expression, and in pUS250 for final expression of a polycistronic construct incorporating PsiD, PsiK, and PsiM. To achieve this, the part was ordered in a gBlock containing BsaI sites (complementary to a BsaI site in the PsiK gBlock and to a BsaI site in the backbone of pUS250) for Golden Gate cloning, and with EcoRI and HindIII sites to enable traditional restriction cloning into pUS250. As there is a ribosomal binding site in the pET-28c(+) backbone, but not in pUS250, a RBS sequence was added to the gBlock upstream of the EcoRI site, so that it would not incorporate into pET-28c(+). The RBS sequence used was the consensus Shine-Dalgarno sequence.
Source
The native sequence for the norbaeocystin methyltransferase PsiM (Psilocybe cubensis) was obtained from NCBI: GenBank accession KY984100.1
References
1. Fricke, J., Blei, F. & Hoffmeister, D. Enzymatic Synthesis of Psilocybin. Angew Chem Int Ed Engl 56, 12352-12355 (2017).