Difference between revisions of "Part:BBa K1373001:Design"
 
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<partinfo>BBa_K1373001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1373001 SequenceAndFeatures</partinfo>
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===Design Notes===
 
===Design Notes===
 
cloning via omega-PCR
 
cloning via omega-PCR
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overexpression intracellular redox state of EAC is one of the most important physiological traits of extracellular electron transfer efficiency.In particular, the NAD+(H) pool size plays a central role of most metabolic pathways. By overexpressing the NAD synthetase, encoded be gene nadE and catalyzes the final step in de novo synthesis and salvage pathway of NAD biosynthesis (Fig. 1), the NAD+ level is increased thereby up-regulating genes whose products catalyze NADH synthesis. Therefore the augmented pool size of NAD+(H) result in promotion of NADH(the carrier of electrons)level, leading to high generation of intracellular releasable electrons and better electricity performance of EAC
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Primers
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F-nadE:ctagctactagagaaagaggagaaaATGACATTGCAACAACAAATAATAAAG
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R-nadE:GATGATTTCTGGAAAAAGTAAtactagtagcggccgctgcagtc
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<html>
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<p></p>
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</html>
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1373000 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K1373000 parameters</partinfo>
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===References===
 
===References===
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1.Yong, X.-Y. et al. Enhancement of bioelectricity generation by cofactor manipulation in microbial fuel cell. Biosensorsand Bioelectronics 56, 19-25, doi:http://dx.doi.org/10.1016/j.bios.2013.12.058 (2014).
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2.Chen L, Wang F, Wang X, Liu YG. (2013) Robust one-tube Ω-PCR strategy accelerates precise sequence modification of plasmids for functional genomics. Plant Cell Physiology

Latest revision as of 07:56, 17 October 2014

nadE with strong promter and strong RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

cloning via omega-PCR

Primers

F-nadE:ctagctactagagaaagaggagaaaATGACATTGCAACAACAAATAATAAAG

R-nadE:GATGATTTCTGGAAAAAGTAAtactagtagcggccgctgcagtc

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Source

the promoter is from iGEM distribution kit and the nadE gene is from the genome of E.coli MG1655

References

1.Yong, X.-Y. et al. Enhancement of bioelectricity generation by cofactor manipulation in microbial fuel cell. Biosensorsand Bioelectronics 56, 19-25, doi:http://dx.doi.org/10.1016/j.bios.2013.12.058 (2014).

2.Chen L, Wang F, Wang X, Liu YG. (2013) Robust one-tube Ω-PCR strategy accelerates precise sequence modification of plasmids for functional genomics. Plant Cell Physiology