Difference between revisions of "Part:BBa K1723001"
 
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<partinfo>BBa_K1723001 short</partinfo>
 
<partinfo>BBa_K1723001 short</partinfo>
  
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PAM rich URS J23117 is a promoter made from the existing J23117 (BBa_J23117) promoter. This promoter has PAM (PAM = NGG sequence) rich Upstream Regulatory Sequence to enable the use of protein dCas9-&#969; (BBa_K1723000) as a gene transcription regulator when in complex with one sgRNA Z0 (BBa_K1723002), Z35 (BBa_K1723003), or Z4 (BBa_K1723004).  
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All this system is operating in the special cell strain JEN202. In our experiments this promoter is 27 base pairs upstream the ATG start codon.  
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<b>PAM rich URS J23117 is an improvement of the J23117 (BBa_J23117) weak promoter</b> [1]. This promoter is the promoter J23117 flanked with a PAM (PAM = NGG sequence) rich Upstream Regulatory Sequence (URS) to enable the use of protein dCas9-&#969; (BBa_K1723000) as a gene transcription regulator when in complex with one sgRNA targeting the promoter such as: the activator sgRNA Z4 (BBa_K1723003), the inhibitor sgRNA Z0 (BBa_K1723002), or the other inhibitor sgRNA Z35 (BBa_K1723004). dCas9 can only bind if the DNA target sequence is preceded by a PAM sequence.  
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<b>This part was experimentally validated, see </b> https://parts.igem.org/Part:BBa_K1723001:Experience
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Discover more parts that can work with this one:
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http://2015.igem.org/Team:EPF_Lausanne/Part_Collection
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https://static.igem.org/mediawiki/2015/f/f9/EPFL_Lausanne_promoter.png
  
 
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<partinfo>BBa_K1723001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1723001 SequenceAndFeatures</partinfo>
  
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===References===
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[1] Bikard, D., Jiang, W., Samai, P., Hochschild, A., Zhang, F., & Marraffini, L. A. (2013). Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic acids research, 41(15), 7429-7437.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
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Latest revision as of 00:47, 19 September 2015

PAM rich URS j23117 promoter


PAM rich URS J23117 is an improvement of the J23117 (BBa_J23117) weak promoter [1]. This promoter is the promoter J23117 flanked with a PAM (PAM = NGG sequence) rich Upstream Regulatory Sequence (URS) to enable the use of protein dCas9-ω (BBa_K1723000) as a gene transcription regulator when in complex with one sgRNA targeting the promoter such as: the activator sgRNA Z4 (BBa_K1723003), the inhibitor sgRNA Z0 (BBa_K1723002), or the other inhibitor sgRNA Z35 (BBa_K1723004). dCas9 can only bind if the DNA target sequence is preceded by a PAM sequence.

This part was experimentally validated, see https://parts.igem.org/Part:BBa_K1723001:Experience

Discover more parts that can work with this one:

http://2015.igem.org/Team:EPF_Lausanne/Part_Collection

EPFL_Lausanne_promoter.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 56
    Illegal NheI site found at 79
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 43
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Bikard, D., Jiang, W., Samai, P., Hochschild, A., Zhang, F., & Marraffini, L. A. (2013). Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic acids research, 41(15), 7429-7437.