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

(→‎Design Notes)
(→‎Design Notes)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
<p><b>Cas9</p></b>
+
<b>Cas9</p>
 
The sequence was ordered as two gBlocks from Integrated DNA Technologies and it was assemble by PvuII restriction enzyme site.
 
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%)
+
<p>• NheI (GCTAGC), which in the alanine GCT codon (18% de codon usage) was replaced by GCG (32%)</p>
−
• EcoRI (C-GAATTC), which in arginine CGA codon (30%) was replaced by CGT (24.7%)
+
<p>• EcoRI (C-GAATTC), which in arginine CGA codon (30%) was replaced by CGT (24.7%) </p>
−
• First HindIII (AAGCTT) changed to AAACTT
+
<p>• First HindIII (AAGCTT) changed to AAACTT</p>
−
• Second HindIII (AAGCTT) changed to AGGCTT
+
<p>• Second HindIII (AAGCTT) changed to AGGCTT</p>
−
• NdeI: CATATG changed to CACATG; CATATG changed to CATACG
+
<p>• NdeI: CATATG changed to CACATG; CATATG changed to CATACG</p>
−
• NcoI: CCATGG changed to CCGTGG
+
<p>• NcoI: CCATGG changed to CCGTGG</p>
−
• BamHI: GGATCC changed to GGACCC
+
<p>• BamHI: GGATCC changed to GGACCC</p>
  
 
===Source===
 
===Source===

Revision as of 01:32, 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
    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</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===