Composite

Part:BBa_K2457006:Design

Designed by: Maria Astolfi, Giovanna Maklouf, Juliana Vitoriano   Group: iGEM17_Amazonas_Brazil   (2017-09-23)
Revision as of 01:34, 2 November 2017 by Giomaklouf (Talk | contribs) (→‎Source)


Cas9 device + Optimized sgRNA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1207
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1042
  • 1000
    INCOMPATIBLE 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