Difference between revisions of "Part:BBa K206000:Design"

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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K206000 short</partinfo>
 
  
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{|
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|<div style="align: center; valign: center; font-family: Arial; font-size: 12pt">
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{|
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|-
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|width="60px"|''Name'':
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|pBAD strong
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|-
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|width="60px"|''Input'':
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|width="100px"|[http://openwetware.org/wiki/Arabinose L-arabinose]
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|-
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|''Output'':
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| PoPS
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|}
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</div>
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<hr width="800px">
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<div class="noprint" style="padding: 10px; color: #ffffff; background-color: #C0C0C0; width: 800px; align: center">
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<center>
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[[Part:BBa_K206000 |Part Main Page]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[Part:BBa_K206000:Design |Part Design]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[Part:BBa_K206000:Characterization |Characterization]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[pBAD Promoter Family |Family]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[Part:BBa_K206000:Experience |Add Data]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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</center>
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</div>
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<br>
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<br>
 
<partinfo>BBa_K206000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K206000 SequenceAndFeatures</partinfo>
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<br>
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==Design Notes==
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Niland et al. found that certain mutations in the AraI1 operator site increased binding of the DNA to the AraC protein [[Part:BBa_K206000:Design#References|[1]]]. We applied all of these mutations with the goal of creating a stronger version of the pBAD promoter. See [[pBAD Promoter Family]] for more details.
  
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==Source==
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Site-directed mutagenesis was performed on <partinfo>I13453</partinfo> using the following primers:<br>
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Forward: 5'-PO4-TAATCTTATGGACTATCTTGCTATGGCATAGC-3'<br>
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Reverse: 5'-PO4-GCGGATCCTACCTGACGCTTTTTATC-3'
  
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===Design Notes===
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''Site-directed mutagenesis:'' We used the Finnzyme Phusion Site-directed Mutagenesis Kit in accordance with the manufacturer's directions.
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Niland et al. found that certain mutations in the AraI1 site increased binding of the DNA to the AraC protein. We applied all of these mutations in an attempt to create a stronger version of the pBAD promoter.
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<br>
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''Primers: ''Phosphorylated oligos were purchased from Integrated DNA Technologies (IDT) and resuspended in water.
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===Source===
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<br>
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Site-directed mutagenesis was performed on <partinfo>I13453</partinfo> using the following primers:<br>
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''Template:'' <partinfo>I13453</partinfo> was obtained from the 2009 iGEM Spring Distribution according to Registry instructions and used as the template for site-directed mutagenesis.
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Forward: TAATCTTATGGACTATCTTGCTATGGCATAGC<br>
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Reverse: GCGGATCCTACCTGACGCTTTTTATC
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===References===
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==References==
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Niland P, Hühne R, Müller-Hill B. (1996). How AraC Interacts Specifically with its Target DNAs. ''J. Mol. Biol.'' 264:667-674. [http://www.ncbi.nlm.nih.gov/pubmed/8980677]
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[http://www.ncbi.nlm.nih.gov/pubmed/8980677 [1]]  Niland P, Hühne R, and Müller-Hill B. (1996). How AraC Interacts Specifically with its Target DNAs. J. Mol. Biol. '''264''':667-674.

Latest revision as of 23:57, 21 October 2009


Name: pBAD strong
Input: [http://openwetware.org/wiki/Arabinose L-arabinose]
Output: PoPS

Part Main Page        Part Design        Characterization        Family        Add Data       




Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Niland et al. found that certain mutations in the AraI1 operator site increased binding of the DNA to the AraC protein [1]. We applied all of these mutations with the goal of creating a stronger version of the pBAD promoter. See pBAD Promoter Family for more details.

Source

Site-directed mutagenesis was performed on BBa_I13453 using the following primers:
Forward: 5'-PO4-TAATCTTATGGACTATCTTGCTATGGCATAGC-3'
Reverse: 5'-PO4-GCGGATCCTACCTGACGCTTTTTATC-3'

Site-directed mutagenesis: We used the Finnzyme Phusion Site-directed Mutagenesis Kit in accordance with the manufacturer's directions.
Primers: Phosphorylated oligos were purchased from Integrated DNA Technologies (IDT) and resuspended in water.
Template: BBa_I13453 was obtained from the 2009 iGEM Spring Distribution according to Registry instructions and used as the template for site-directed mutagenesis.

References

[http://www.ncbi.nlm.nih.gov/pubmed/8980677 [1]] Niland P, Hühne R, and Müller-Hill B. (1996). How AraC Interacts Specifically with its Target DNAs. J. Mol. Biol. 264:667-674.