Difference between revisions of "Part:BBa K1723003:Design"

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__NOTOC__
 
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<partinfo>BBa_K1723003 short</partinfo>
 
<partinfo>BBa_K1723003 short</partinfo>
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===Design Notes===
 
===Design Notes===
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This part was design on the model for sgRNAs on the paper from Alec AK Nielsen & Christopher A Voigt [3] and the specific sequence for the gRNA Z4 was designed from the sequence of the plasmid pWJ89 D.Bikard [1] sent us.
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This part was design on the model for sgRNAs on the paper from Alec AK Nielsen & Christopher A Voigt [2] and the specific sequence for the gRNA Z4 was designed from the sequence of the PAM rich URS J23117 (BBa_K1723001) promoter of the plasmid pWJ89 D.Bikard [1] sent us.
  
 
this part contains a deletion, the 15th base of the terminator is missing. The original sequence is ATTGCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTT. All of our tests were performed with the deletion and it didn't affect the sgRNA function.
 
this part contains a deletion, the 15th base of the terminator is missing. The original sequence is ATTGCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTT. All of our tests were performed with the deletion and it didn't affect the sgRNA function.
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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.
 
[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.
  
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[2] Qi, L. S., Larson, M. H., Gilbert, L. A., Doudna, J. A., Weissman, J. S., Arkin, A. P., & Lim, W. A. (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell, 152(5), 1173-1183.
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[2] Alec AK Nielsen & Christopher A Voigt (2014). Multi-input CRISPR/Cas circuits that interface host regulatory network. Molecular systems biology, 10(11), 763.
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[3] Alec AK Nielsen & Christopher A Voigt (2014). Multi-input CRISPR/Cas circuits that interface host regulatory network. Molecular systems biology, 10(11), 763.
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Latest revision as of 17:55, 18 September 2015

sgRNA Z4 expressing cassette


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 185
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 185
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 126
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 185
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 185
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was design on the model for sgRNAs on the paper from Alec AK Nielsen & Christopher A Voigt [2] and the specific sequence for the gRNA Z4 was designed from the sequence of the PAM rich URS J23117 (BBa_K1723001) promoter of the plasmid pWJ89 D.Bikard [1] sent us.

this part contains a deletion, the 15th base of the terminator is missing. The original sequence is ATTGCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTT. All of our tests were performed with the deletion and it didn't affect the sgRNA function.

Source

This sequence was fully synthesized.

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.

[2] Alec AK Nielsen & Christopher A Voigt (2014). Multi-input CRISPR/Cas circuits that interface host regulatory network. Molecular systems biology, 10(11), 763.