Part:BBa_K3102045:Design

Synthesis and externalization of lactate (DLD-II Operon, MtrC)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 4365
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4949
  

Design Notes

We mutated the MtrC sequence with a Guanine into Cysteine and a Guanine into Adenine at the 1795bp and 1798bp position (G1795C & G1798A), respectively, because of the PstI enzyme (BBa_K3102020).

The DLD-II operon sequence was mutated as followed :

- a Cysteine into Thymine and a Guanine into Adenine at the 428bp and 433bp position (C428T & G433A), respectively, because of the PstI enzyme.

- a Adenine into Guanine and a Cysteine into Thymine at the 2799bp and 2802bp position (A2799G & C2802T), respectively, because of the EcoRI enzyme. (BBa_K3102019)

Source

DLD-II: sequence comes from Shewanella Oneidensis was found through Biocyc: https://biocyc.org/gene?orgid=SONE211586&id=G1GMP-1405-MONOMER#tab=TU

MtrC: sequence comes from Shewanella Oneidensis was found through Biocyc: https://biocyc.org/gene?orgid=SONE211586&id=G1GMP-1627

Promoter (BBa_I719005), RBS (BBa_B0034) and Terminator (BBa_B0015) are from the iGEM Registry.

References

Kouzuma A, Kasai T, Hirose A, Watanabe K. "Catabolic and regulatory systems in shewanella oneidensis MR-1 involved in electricity generation in microbial fuel cells". Front Microbiol. 2015;6(JUN):1–11.