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, an average of 30bp is recommended.  
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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.