Difference between revisions of "Part:BBa K3192025:Design"

 
 
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===Design Notes===
 
===Design Notes===
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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>
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+
  
  
 
===Source===
 
===Source===
  
−
Pseudomonas putida  
+
<i>Pseudomonas putida </i>
−
Escherichia coli
+
<br>
 +
<i>Escherichia coli </i>
 +
<br>
 
Synthetic  
 
Synthetic  
  
 
===References===
 
===References===
 +
<p><i> Pseudomonas putida</i> strain SN1 StyA (styA), StyB (styB), StyC (styC), a - Nucleotide - NCBI. (n.d.). Retrieved from https://www.ncbi.nlm.nih.gov/nuccore/DQ177365.1.</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>
 +
 +
<p> <i>Pseudomonas putida influx</i> porin (styE) gene, complete cds - Nucleotide - NCBI. (n.d.). Retrieved from https://www.ncbi.nlm.nih.gov/nuccore/AY450871.1</p>

Latest revision as of 02:53, 22 October 2019


genes styABCDE


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2440
    Illegal NheI site found at 5360
    Illegal NheI site found at 5383
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1062
    Illegal BamHI site found at 3841
    Illegal BamHI site found at 5017
    Illegal XhoI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 143
    Illegal NgoMIV site found at 3001
    Illegal NgoMIV site found at 3368
    Illegal AgeI site found at 3191
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 3779


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

Pseudomonas putida
Escherichia coli
Synthetic

References

Pseudomonas putida strain SN1 StyA (styA), StyB (styB), StyC (styC), a - Nucleotide - NCBI. (n.d.). Retrieved from https://www.ncbi.nlm.nih.gov/nuccore/DQ177365.1.

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).

Pseudomonas putida influx porin (styE) gene, complete cds - Nucleotide - NCBI. (n.d.). Retrieved from https://www.ncbi.nlm.nih.gov/nuccore/AY450871.1