Difference between revisions of "Part:BBa K3192031:Design"

 
 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
na
+
<p>This composite part was assembled using Golden Gate Assembly. The 2019 Virginia iGEM team designed these composite parts for assembly using Teselagen Software. Each coding sequence was codon optimized for <i>E. coli</i> K12 strains. The DNA was synthesized in fragments ranging from 1-1.4kb, containing BsaI restriction sites required for Golden Gate Assembly and adjacent 5 base pair complementary overlaps for assembly specificity. The sequences were checked for illegal sites and conservative base pair substitutions were made as necessary. The composite part was then assembled and sequenced for identity confirmation.</p>
−
 
+
  
  
 
===Source===
 
===Source===
  
−
C. nectar
+
<i>Cupriavidus necator </i>
−
E. coli  
+
<br>
 +
<i>Escherichia coli </i>
 +
<br>
 
Synthetic  
 
Synthetic  
  
 
===References===
 
===References===
 +
<p> <i>Cupriavidus necator</i> H16 PhaC1 (phaC1), PhaA (phaA), and PhaB1 (phaB1). (n.d.). Retrieved from https://www.ncbi.nlm.nih.gov/nuccore/MH558939.1.</p>
 +
 +
<p> Sclavi, B. <i>et al</i>. Real-time characterization of intermediates in the pathway to open complex formation by <i>Escherichia coli</i> RNA polymerase at the T7A1 promoter. <i>Proceedings of the National Academy of Sciences </i> 13, 4706–4711 (2005). </p>

Latest revision as of 02:59, 22 October 2019


pha plasmid with neokanamycin resistance


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1249
    Illegal NheI site found at 3712
    Illegal NheI site found at 3735
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2354
    Illegal BamHI site found at 394
    Illegal BamHI site found at 2646
    Illegal BamHI site found at 3216
    Illegal XhoI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 354
    Illegal NgoMIV site found at 4240
    Illegal AgeI site found at 2853
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 4089
    Illegal SapI.rc site found at 4299


Design Notes

This composite part was assembled using Golden Gate Assembly. The 2019 Virginia iGEM team designed these composite parts for assembly using Teselagen Software. Each coding sequence was codon optimized for E. coli K12 strains. The DNA was synthesized in fragments ranging from 1-1.4kb, containing BsaI restriction sites required for Golden Gate Assembly and adjacent 5 base pair complementary overlaps for assembly specificity. The sequences were checked for illegal sites and conservative base pair substitutions were made as necessary. The composite part was then assembled and sequenced for identity confirmation.


Source

Cupriavidus necator
Escherichia coli
Synthetic

References

Cupriavidus necator H16 PhaC1 (phaC1), PhaA (phaA), and PhaB1 (phaB1). (n.d.). Retrieved from https://www.ncbi.nlm.nih.gov/nuccore/MH558939.1.

Sclavi, B. et al. Real-time characterization of intermediates in the pathway to open complex formation by Escherichia coli RNA polymerase at the T7A1 promoter. Proceedings of the National Academy of Sciences 13, 4706–4711 (2005).