Difference between revisions of "Part:BBa K2570001:Design"

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We ordered the part DNA from a synthesis company and had codon optimization of the sequence.
 
We ordered the part DNA from a synthesis company and had codon optimization of the sequence.
 
Mutation of restriction sites EcoRⅠ, SpeⅠand PstⅠ.
 
Mutation of restriction sites EcoRⅠ, SpeⅠand PstⅠ.
 +
 +
Mutation 1: Aro10-Pst1 （Pro 619-621）cctgca-ccggca
 +
 +
Sense primer: TATATTTTTGTTCCGGCAGATTTTG
 +
 +
Anti-sense primer: AATCTGCCGGAACAAAAATATAACC
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 +
Mutation 2: Aro10-Spe1 （Thr 741-744）actagt- accagt
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 +
Sense primer: ACAAGATTACCAGTTGGATATATTC
 +
 +
Anti-sense primer: ATATATCCAACTGGTAATCTTGTTG
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Mutation 3: Aro10-EcoR1（Asn 1791-1794）gaattc-gaactc
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Sense primer: CTTAAGAACTCAAACAAAAGAAGCG
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Anti-sense primer: TTTGTTTGAGTTCTTAAGCTCCTCC
  
 
===Source===
 
===Source===

Revision as of 09:22, 5 October 2018


Aro10


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

We ordered the part DNA from a synthesis company and had codon optimization of the sequence. Mutation of restriction sites EcoRⅠ, SpeⅠand PstⅠ.

Mutation 1: Aro10-Pst1 （Pro 619-621）cctgca-ccggca

Sense primer: TATATTTTTGTTCCGGCAGATTTTG

Anti-sense primer: AATCTGCCGGAACAAAAATATAACC

Mutation 2: Aro10-Spe1 （Thr 741-744）actagt- accagt

Sense primer: ACAAGATTACCAGTTGGATATATTC

Anti-sense primer: ATATATCCAACTGGTAATCTTGTTG

Mutation 3: Aro10-EcoR1（Asn 1791-1794）gaattc-gaactc

Sense primer: CTTAAGAACTCAAACAAAAGAAGCG

Anti-sense primer: TTTGTTTGAGTTCTTAAGCTCCTCC

Source

The phenylpyruvate decarboxylase from S. cerevisiae (Aro10, NP 010668.3).

https://www.ncbi.nlm.nih.gov/protein/NP_010668.3?report=genbank&log$=protalign&blast_rank=1&RID=V0V04K3B015

References

Wang, P., Yang, X., Lin, B., Huang, J., & Tao, Y. (2017). Cofactor self-sufficient whole-cell biocatalysts for the production of 2-phenylethanol. Metabolic Engineering, 44(August), 143–149.