Difference between revisions of "Part:BBa K203110:Design"
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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]] | + | [[Image:Rapcr.jpg|thumb|none|600px]] |
===Source=== | ===Source=== | ||
Revision as of 08:37, 16 October 2009
Constitutive promoter; 0.4 REU
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
Source
To be updated