Difference between revisions of "Part:BBa K2457005:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | <b>Cas9</ | + | <b>Cas9</b> |
The sequence was ordered as two gBlocks from Integrated DNA Technologies and it was assemble by PvuII restriction enzyme site. | 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>• NheI (GCTAGC), which in the alanine GCT codon (18% de codon usage) was replaced by GCG (32%)</p> | ||
Revision as of 01:33, 2 November 2017
Standardized pBAD_Cas9 device
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1207
- 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
descrever