Part:BBa_K3192029:Design

sty plasmid with neokanamycin resistance

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 2490
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1076
  Illegal BamHI site found at 3897
  Illegal BamHI site found at 5087
  Illegal XhoI site found at 1
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 157
  Illegal NgoMIV site found at 3057
  Illegal NgoMIV site found at 3424
  Illegal AgeI site found at 3247
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4172
  Illegal BsaI.rc site found at 1935
  Illegal SapI site found at 3835
  

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.

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