Coding

Part:BBa_K2457001:Design

Designed by: Maria Astolfi, Giovanna Maklouf, Juliana Vitoriano   Group: iGEM17_Amazonas_Brazil   (2017-07-09)
Revision as of 01:29, 2 November 2017 by Giomaklouf (Talk | contribs) (→‎Design Notes)


Standardized Cas9 coding sequence + B0015 terminator


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Unknown
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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