Coding

Part:BBa_K3726065:Design

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-07)


CDS_AcrbV2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 331
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 60
    Illegal PstI site found at 331
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 331
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 331
    Illegal NgoMIV site found at 1467
    Illegal NgoMIV site found at 2983
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part corresponds with the codon optimized coding sequence of AcrbV2. Codon optimization has been performed using DeNovo DNA Software, adjusting the codon usage for high expression in Synechococcus elongatus PCC7942 . In addition the part sequence has been optimized to improve mRNA stability removing internal recognition sites for endonucleases

Source

Coding sequence of this membrane transporter subunit comes from directed evolution of AcrB Escherichia coli (strain K12) gen. Uniprot reference: https://www.uniprot.org/uniprot/P31224

References

"Enhancing Tolerance to Short-Chain Alcohols by Engineering the Escherichia coli AcrB Efflux Pump to Secrete the Non-native Substrate n-Butanol", ACS Publications, 2021. [Online]. Available: https://pubs.acs.org/doi/10.1021/sb400065q.