Difference between revisions of "Part:BBa K676002:Design"

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===Design Notes===
 
===Design Notes===
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The mNARK promoter used was from the UCL 2009 Stress Light device
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First Stage: Performed 3A assembly between mNark promoter ([[Part:BBa_K239006]]) and RBS ([[Part:BBa_B0034]]) and pSB1K3 plasmid backbone.
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Performed 3A assembly between YFP.LVA ([[Part:BBa_E0032]]) and terminator ([[Part:BBa_B0012]]) and pSB1T3 plasmid backbone.
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Second Stage: Performed 3A assembly between mNark promoter/RBS ligation ([[Part:BBa_S04716]]) and YFP.LVA/Ter ligation ([[Part:BBa_S04719]]) and pSB1C3 plasmid backbone.
  
 
[[Image:Stress_Lights_2.0_Experiment.jpg]]
 
[[Image:Stress_Lights_2.0_Experiment.jpg]]
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===Source===
 
===Source===
  
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mNarK promoter is designed to have better affinity to activated Fnr than the NarK promoter (BBa_K239005) and hence also stronger activity under anaerobic conditions. It is also shorter than the NarK promoter (BBa_K239005).
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All the parts used to construct this device was obtained from the registry.
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===References===
 
===References===
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1) Kolesnikow, T., I. Schroder, and R.P. Gunsalus, Regulation of narK gene expression in Escherichia coli in response to anaerobiosis, nitrate, iron, and molybdenum. J Bacteriol, 1992. 174(22): p. 7104-11.
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2) Unden, G. and M. Trageser, Oxygen regulated gene expression in Escherichia coli: control of anaerobic respiration by the FNR protein. Antonie Van Leeuwenhoek, 1991. 59(2): p. 65-76.
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3) Spiro, S. and J.R. Guest, FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev, 1990. 6(4): p. 399-428.

Latest revision as of 21:45, 9 October 2011

mNARK with YFP.LVA


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]


Design Notes

First Stage: Performed 3A assembly between mNark promoter (Part:BBa_K239006) and RBS (Part:BBa_B0034) and pSB1K3 plasmid backbone. Performed 3A assembly between YFP.LVA (Part:BBa_E0032) and terminator (Part:BBa_B0012) and pSB1T3 plasmid backbone.

Second Stage: Performed 3A assembly between mNark promoter/RBS ligation (Part:BBa_S04716) and YFP.LVA/Ter ligation (Part:BBa_S04719) and pSB1C3 plasmid backbone.

Stress Lights 2.0 Experiment.jpg

Source

All the parts used to construct this device was obtained from the registry.

References

1) Kolesnikow, T., I. Schroder, and R.P. Gunsalus, Regulation of narK gene expression in Escherichia coli in response to anaerobiosis, nitrate, iron, and molybdenum. J Bacteriol, 1992. 174(22): p. 7104-11.

2) Unden, G. and M. Trageser, Oxygen regulated gene expression in Escherichia coli: control of anaerobic respiration by the FNR protein. Antonie Van Leeuwenhoek, 1991. 59(2): p. 65-76.

3) Spiro, S. and J.R. Guest, FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev, 1990. 6(4): p. 399-428.