Difference between revisions of "Part:BBa K2457005:Design"

 
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===Design Notes===
 
===Design Notes===
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descrever
+
<p><b>Cas9</p></b>
−
 
+
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===
 
===Source===

Revision as of 01:31, 2 November 2017


Standardized pBAD_Cas9 device


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1207
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 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</p> 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

===References===