Difference between revisions of "Part:BBa K2457001:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
−
Assembly by Gibson/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>• 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===

Revision as of 01:29, 2 November 2017


Standardized Cas9 coding sequence + B0015 terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 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