Difference between revisions of "Part:BBa K3102046:Design"

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===Design Notes===
 
===Design Notes===
−
The ACS sequence is mutated a Cytosine into Guanine at the 1609bp position (C1609T) because of the Esp3I enzyme that we used for our Golden Gate Assembly (GGA) construct. (e.g. BBa_K3102022).
+
The ACS sequence is mutated a Cytosine into Guanine at the 1566bp position (C1566G) because of the Esp3I enzyme that we used for our Golden Gate Assembly (GGA) construct. (BBa_K3102022).
  
−
The MDH sequence is mutated an Adenine into Guanine at the 829bp position (A829G) because of the EcoRI enzyme. (e.g. BBa_K3102018)
+
The MDH sequence is mutated an Adenine into Guanine at the 786bp position (A786G) because of the EcoRI enzyme. (BBa_K3102018)
  
 
===Source===
 
===Source===

Latest revision as of 10:24, 16 October 2019


Electrons production (pflB, ACS, FDH, MDH)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6180
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 5516
    Illegal AgeI site found at 534
    Illegal AgeI site found at 3413
    Illegal AgeI site found at 6024
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 836


Design Notes

The ACS sequence is mutated a Cytosine into Guanine at the 1566bp position (C1566G) because of the Esp3I enzyme that we used for our Golden Gate Assembly (GGA) construct. (BBa_K3102022).

The MDH sequence is mutated an Adenine into Guanine at the 786bp position (A786G) because of the EcoRI enzyme. (BBa_K3102018)

Source

PFLB: sequence comes from Shewanella Oneidensis and was found through Biocyc: https://biocyc.org/gene?orgid=SONE211586&id=G1GMP-2686

ACS: sequence comes from Shewanella Oneidensis and was found through Biocyc: https://biocyc.org/gene?orgid=SONE211586&id=G1GMP-2522#tab=TU

FDH: sequence comes from Saccharomyces cerevisiae was found through Yeastgenome: https://www.yeastgenome.org/locus/S000005915

MDH: sequence comes from Shewanella Oneidensis and was found through Biocyc: https://biocyc.org/gene?orgid=SONE211586&id=G1GMP-721

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

References

Brown TD. et al., The enzymic interconversion of acetate and acetyl-coenzyme A in Escherichia coli, J Gen Microbiol. 1977 Oct;102(2):327-36.

Kumari S. et al., Cloning, characterization, and functional expression of acs, the gene which encodes acetyl coenzyme A synthetase in Escherichia coli, J Bacteriol. 1995 May;177(10):2878-86.

Li F, Li YX, Cao YX, Wang L, Liu CG, Shi L, et al. Modular engineering to increase intracellular NAD(H/+) promotes rate of extracellular electron transfer of Shewanella oneidensis. Nat Commun [Internet]. 2018;9(1):1–13. Available from: http://dx.doi.org/10.1038/s41467-018-05995-8

Sutherland P. et al., "Isolation and expression of the Escherichia coli gene encoding malate dehydrogenase.", J. Bacteriology, 163:1074-1079(1985)

Zhenquan Lin et al., “Metabolic engineering of Escherichia coli for the production of riboflavin, Microbial Cell Factories, 2014, 13:104.