Difference between revisions of "Part:BBa K3102044:Design"
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AWS: E.coli, sequence found through Biocyc https://biocyc.org/gene?orgid=ECOLI&id=EG11448 | AWS: E.coli, sequence found through Biocyc https://biocyc.org/gene?orgid=ECOLI&id=EG11448 | ||
Revision as of 08:34, 16 October 2019
Electron transfer improvement (ACS, ZWF, GND, PGL)
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2933
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 2933
Illegal NotI site found at 2814 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3490
Illegal XhoI site found at 408 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2933
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2933
Illegal NgoMIV site found at 2206
Illegal NgoMIV site found at 2923
Illegal NgoMIV site found at 5493
Illegal AgeI site found at 837
Illegal AgeI site found at 4245 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 3975
Design Notes
In the ACS, we mutated a Cytosine into Thymine at the 1563bp position (C1563T) because of the Esp3I enzyme that we used for our Golden Gate Assembly (GGA) construct. (e.g BBa_K3102021)
In the ZWF, we mutated a Thymine into Cysteine at the 844bp position (T844C) because of the PstI enzyme (e.g. BBa_K3102008)
Source
AWS: E.coli, sequence found through Biocyc https://biocyc.org/gene?orgid=ECOLI&id=EG11448
ZWF: Corynebacterium glutamicum, sequence found through Ebi https://www.ebi.ac.uk/ena/data/view/BAB98969
GND: Corynebacterium glutamicum, sequence found through GenBank https://biocyc.org/gene?orgid=CORYNE&id=G18NG-11035-MONOMER#tab=TU
GLD: E.coli, sequence found through GenBank https://www.ncbi.nlm.nih.gov/nuccore/NC_000913.3?from=798586&to=799581&report=fasta
Promoter (BBa_I719005), RBS (BBa_B0034) and Terminator (BBa_B0015) are from the iGEM Registry.
References
Bacher A. et al., "Riboflavin synthases of Bacillus subtilis. Purification and properties." Biol. Chem. 255:632-637(1980)
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.
Chen Y.Y. et al., "Conformational changes associated with cofactor/substrate binding of 6-phosphogluconate dehydrogenase from Escherichia coli and Klebsiella pneumoniae: Implications for enzyme mechanism.", J. Struct. Biol. 169:25-35(2010)
Eberhardt S.M.R. et al., J"Cloning, sequencing, mapping and hyperexpression of the ribC gene coding for riboflavin synthase of Escherichia coli." . Biochem. 242:712-719(1996)
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.
Lin Z, et al., "Metabolic engineering of Escherichia coli for the production of riboflavin". Microb Cell Fact. 2014;13(1):1–12.
Thomason L.C., Court D.L. et al., "Identification of the Escherichia coli K-12 ybhE gene as pgl, encoding 6-phosphogluconolactonase." Bacteriol. 186:8248-8253(2004)
Zhao J. et al., "Effect of zwf gene knockout on the metabolism of Escherichia coli grown on glucose or acetate." Metab. Eng. 6:164-174(2004)
Zhenquan Lin et al., “Metabolic engineering of Escherichia coli for the production of riboflavin," Microbial Cell Factories, 2014, 13:104.