Difference between revisions of "Part:BBa K3352001:Design"

 
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===Design Notes===
 
===Design Notes===
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We attached a 6x his-tag upstream of the phi29 DNA Polymerase for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF.  
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We obtained the amino acid sequence of Φ29 DNA polymerase from <i>Bacillus phage Φ29</i>. We optimized the DNA sequence for expression in <i>E. coli</i> and removed the EcoRI cutting site to conform to BioBrick assembly standards.
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Revision as of 06:37, 24 October 2020


Φ29 DNA Polymerase with His-Tag and GS linker Sequence


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 595
    Illegal SapI.rc site found at 993


Design Notes

We obtained the amino acid sequence of Φ29 DNA polymerase from Bacillus phage Φ29. We optimized the DNA sequence for expression in E. coli and removed the EcoRI cutting site to conform to BioBrick assembly standards.


Source

We obtained the amino acid sequence of phi29 DNA polymerase from Bacillus phage phi29.

References