Part:BBa_K3102046:Design
Electrons production (pflB, ACS, FDH, MDH)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 6180
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 836
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 MDH sequence is mutated an Adenine into Guanine at the 829bp position (A829G) because of the EcoRI enzyme. (e.g. 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.