Composite

Part:BBa_M36659:Design

Designed by: Taylor Marie Chavez, Maria Iglesias, Alison Jahansouz   Group: Stanford BIOE44 - S11   (2015-10-23)


Poly-3-hydroxybutyrate Growth Actuator


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 250
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 250
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 246
    Illegal XhoI site found at 1369
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 250
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 250
    Illegal AgeI site found at 1405
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Design considerations include avoiding BbsI, BsaI, and BsmBI and ensuring less than five repeats of 10 bp. Restriction sites were avoided to not conflict with DNA 2.0 synthesis process, as was the minimization of repeats in the sequence.

Source

Xanthomonas campestris pv. raphani 756C is the organism from which the amino acid sequence for poly-beta-hydroxybutyrate polymerase was found and Phaeobacter inhibens (strain ATCC 700781/DSM 17395/CIP 105210/NBRC 16654/BS107) is the organism from which the amino acid sequence for acetoacetyl-CoA reductase was found. In both cases, the amino acid sequence was obtained and the DNA sequences were generated.

References include http://www.uniprot.org/uniprot/G0CFI5, http://www.uniprot.org/uniprot/I7EIV2, and http://www.google.com/patents/WO1995020621A1?cl=en.

DNA sequence was generated from IDT Codon Optimization in E. coli K12.

References

Referenced http://www.uniprot.org/uniprot/G0CFI5, http://www.uniprot.org/uniprot/I7EIV2, and http://www.google.com/patents/WO1995020621A1?cl=en, which all contributed to the generation of the gene sequence for the composite.