Difference between revisions of "Part:BBa K2457005:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | + | <b>Cas9</b> | |
− | + | The sequence was ordered as two gBlocks from Integrated DNA Technologies and it was assemble by PvuII restriction enzyme site. | |
− | + | <p>• NheI (GCTAGC), which in the alanine GCT codon (18% de codon usage) was replaced by GCG (32%)</p> | |
+ | <p>• EcoRI (C-GAATTC), which in arginine CGA codon (30%) was replaced by CGT (24.7%) </p> | ||
+ | <p>• First HindIII (AAGCTT) changed to AAACTT</p> | ||
+ | <p>• Second HindIII (AAGCTT) changed to AGGCTT</p> | ||
+ | <p>• NdeI: CATATG changed to CACATG; CATATG changed to CATACG</p> | ||
+ | <p>• NcoI: CCATGG changed to CCGTGG</p> | ||
+ | <p>• BamHI: GGATCC changed to GGACCC</p> | ||
===Source=== | ===Source=== | ||
− | + | <b>Cas9</b> M1GAS Streptococcus pyogenes | |
− | + | ||
===References=== | ===References=== | ||
+ | <p>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.</p> | ||
+ | <p>Church, George M.; Mali, Prashant; Esvelt, Kevin M. 2013. Cas9 as a versatile tool for engineering biology. Nature Methods, 10: 957-963.</p> |
Latest revision as of 01:46, 2 November 2017
Standardized pBAD_Cas9 device
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1042
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1024
Design Notes
Cas9 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
Cas9 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.