Composite
Part:BBa_K2457006:Design
Designed by: Maria Astolfi, Giovanna Maklouf, Juliana Vitoriano Group: iGEM17_Amazonas_Brazil (2017-09-23)
Cas9 device + Optimized sgRNA
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