Difference between revisions of "Part:BBa K2457001:Design"
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===References=== | ===References=== | ||
+ | Nishimasu, Hiroshi; Hsu, Patrick D.; Shehata, Soraya; Nureki, Osamu. et al. 2014. Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell, 156(5): 935–949. | ||
+ | Church, George M.; Mali, Prashant; Esvelt, Kevin M. 2013. Cas9 as a versatile tool for engineering biology. Nature Methods, 10: 957-963. |
Revision as of 01:45, 2 November 2017
Standardized Cas9 coding sequence + B0015 terminator
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sequence was ordered as two gBlocks from Integrated DNA Technologies and it was assemble by PvuII restriction enzyme site.
• NheI (GCTAGC), which in the alanine GCT codon (18% de codon usage) was replaced by GCG (32%)
• EcoRI (C-GAATTC), which in arginine CGA codon (30%) was replaced by CGT (24.7%)
• First HindIII (AAGCTT) changed to AAACTT
• Second HindIII (AAGCTT) changed to AGGCTT
• NdeI: CATATG changed to CACATG; CATATG changed to CATACG
• NcoI: CCATGG changed to CCGTGG
• BamHI: GGATCC changed to GGACCC
Source
M1GAS Streptococcus pyogenes
References
Nishimasu, Hiroshi; Hsu, Patrick D.; Shehata, Soraya; Nureki, Osamu. et al. 2014. Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell, 156(5): 935–949. Church, George M.; Mali, Prashant; Esvelt, Kevin M. 2013. Cas9 as a versatile tool for engineering biology. Nature Methods, 10: 957-963.