Difference between revisions of "Part:BBa K2043001:Design"
 
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===Design Notes===
 
===Design Notes===
−
His-tag is added in the C-terminal for optimised protein purification
+
His-tag is added in the C-terminal for protein purification<br>
−
<i>catA</i> was codon optimised for <i>E. coli</i>.
+
The sequence was codon optimized for E. coli and restriction sites for the restriction enzymes BpiI, BsaI and BsmBI were avoided.<br>
−
 
+
The sequence was confirmed by sequencing and no mutations were observed.<br>
−
 
+
The construction was done by Golden Gate technique using BsaI as restriction enzyme to obtain the desired cohesive ends.
  
 
===Source===
 
===Source===
  
−
<i>Acinetobacter pittii</i>'s genome
+
<i>Acinetobacter pittii</i>'s genome,
 +
NCBI Reference Sequence for the gene: YP_004995593.1
  
 
===References===
 
===References===
 +
Lin, J., & Milase, R. N. (2015). Purification and Characterization of Catechol 1, 2-Dioxygenase from Acinetobacter sp. Y64 Strain and Escherichia coli Transformants. The protein journal, 34(6), 421-433. <br><br>

Latest revision as of 02:01, 27 October 2016


catA from Acinetobacter pittii, codon optimized for E. coli


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
    COMPATIBLE WITH RFC[1000]


Design Notes

His-tag is added in the C-terminal for protein purification
The sequence was codon optimized for E. coli and restriction sites for the restriction enzymes BpiI, BsaI and BsmBI were avoided.
The sequence was confirmed by sequencing and no mutations were observed.
The construction was done by Golden Gate technique using BsaI as restriction enzyme to obtain the desired cohesive ends.

Source

Acinetobacter pittii's genome, NCBI Reference Sequence for the gene: YP_004995593.1

References

Lin, J., & Milase, R. N. (2015). Purification and Characterization of Catechol 1, 2-Dioxygenase from Acinetobacter sp. Y64 Strain and Escherichia coli Transformants. The protein journal, 34(6), 421-433.