Difference between revisions of "Part:BBa K5325000:Design"

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===Design Notes===
 
===Design Notes===
−
Addition of a FLAG tag that aids protein purification and visualization on Western Blot.<br>
+
Addition of a C-terminal FLAG tag that aids protein purification and visualization on Western Blot.<br>
−
DNA sequence was codon optimized for expression in <i>E. coli</i>, which included the change in the start codon from GTG to ATG.
+
Original DNA sequence was codon optimized for expression in <i>E. coli</i> and <i>Shewanella oneidensis</i> MR-1.
  
 
===Source===
 
===Source===
  
−
The sequence was taken from <i>Alcanivorax borkumensis</i> genome
+
The part originated from <i>Alcanivorax borkumensis</i> SK2 sequence, which is an identical sequence to the PLA depolymerase DNA sequence mentioned in the referenced study.
 +
<br>NCBI ID and link to NCBI profile: <HTML><a href="https://www.ncbi.nlm.nih.gov/nuccore/NC_008260.1?from=2773448&to=2774365&report=genbank">NC_008260.1</a></html>
  
 
===References===
 
===References===
 +
Hajighasemi, M., Nocek, B.P., <i>et al.</i> (2016). "Biochemical and Structural Insights into Enzymatic Depolymerization of Polylactic Acid and Other Polyesters by Microbial Carboxylesterases." Biomacromolecules 17(6):2027-2039

Latest revision as of 02:48, 2 October 2024


ABO_plaA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 757
    Illegal AgeI site found at 673
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Addition of a C-terminal FLAG tag that aids protein purification and visualization on Western Blot.
Original DNA sequence was codon optimized for expression in E. coli and Shewanella oneidensis MR-1.

Source

The part originated from Alcanivorax borkumensis SK2 sequence, which is an identical sequence to the PLA depolymerase DNA sequence mentioned in the referenced study.
NCBI ID and link to NCBI profile: NC_008260.1

References

Hajighasemi, M., Nocek, B.P., et al. (2016). "Biochemical and Structural Insights into Enzymatic Depolymerization of Polylactic Acid and Other Polyesters by Microbial Carboxylesterases." Biomacromolecules 17(6):2027-2039