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

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===Design Notes===
 
===Design Notes===
  
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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.
+
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.
  
 
[[Image:Rapcr.jpg|thumb|none|600px]]
 
[[Image:Rapcr.jpg|thumb|none|600px]]

Revision as of 08:37, 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 [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.

Rapcr.jpg

Source

To be updated

References