Part:BBa_K2570001:Design

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.

Site-directed mutation site <style="font-weight:bold;">

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.