Difference between revisions of "Part:BBa K203110:Design"

 
(Design Notes)
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__NOTOC__
 
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<partinfo>BBa_K203110 short</partinfo>
 
<partinfo>BBa_K203110 short</partinfo>
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===Design Notes===
 
===Design Notes===
To be updated
 
  
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We performed [[http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters|RA-PCR]] with oligos containing binding sites for some well known generally activating transcription factors (Sp1, Ap1, CREB, NF-Y) which we identified from literature search [1],[2],[3]. We also added NF-κB responsive oligos as NF-κB has non-specific activity and is therefore used by a variety of viral constitutive promoters, e.g. the HIV promoter [4]. We picked 24 colonies, two of which we dismissed after a test digest. We then cloned the construct into [[Part:BBa_K203112]] by SpeI and HindIII to obtain a core promoter.
  
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[[Image:Rapcr.jpg]]
  
 
===Source===
 
===Source===

Revision as of 08:36, 16 October 2009

Constitutive promoter; 0.4 REU


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

We performed RA-PCR with oligos containing binding sites for some well known generally activating transcription factors (Sp1, Ap1, CREB, NF-Y) which we identified from literature search [1],[2],[3]. We also added NF-κB responsive oligos as NF-κB has non-specific activity and is therefore used by a variety of viral constitutive promoters, e.g. the HIV promoter [4]. We picked 24 colonies, two of which we dismissed after a test digest. We then cloned the construct into Part:BBa_K203112 by SpeI and HindIII to obtain a core promoter.

Rapcr.jpg

Source

To be updated

References