Difference between revisions of "Part:BBa K1699005:Design"

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===Design Notes===
 
===Design Notes===
  
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gRNA in SaCas9 scaffold under U6 promoter. Designed to complement 3 sites on MLP synthetic promoter.
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Designed using Benchling to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restriction sites.
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<br /> Cloned into pSBC13 using EcoRI and PstI restriction sites.
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<br /> F. tagtcatggcggccgcgtcgacG
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<br /> R. actgacatgcggccgcttaattaaC
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===Source===
 
===Source===
  
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gMLP from IDT de-novo synthesis. Cloning with U6 to pSBC13 was done by team BGU 2015.
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Sequence of gRNA:
  
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===References===
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Tunable and multifunctional eukaryotic transcription factors based on CRISPR/Cas. Farzadfard F, Perli SD, Lu TK. ACS Synth Biol. 2013 Oct 18;2(10):604-13. doi: 10.1021/sb400081r. Epub 2013 Sep 11. http://www.ncbi.nlm.nih.gov/pubmed/23977949
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1. In vivo genome editing using Staphylococcus aureus Cas9.  
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<br />http://10.1038/nature14299  
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Sequence of U6 promoter:
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2. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
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<br />http://pubs.acs.org/doi/full/10.1021/sb400081r
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In vivo genome editing using Staphylococcus aureus Cas9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. Nature. 2015 Apr 9;520(7546):186-91. doi: 10.1038/nature14299. Epub 2015 Apr 1. http://www.ncbi.nlm.nih.gov/pubmed/25830891

Latest revision as of 11:38, 13 September 2015

gRNA for dCas9-VP64 targeting synthetic activation promoter pMLPm under U6 promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 250
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Designed using Benchling to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restriction sites.

Source

Sequence of gRNA:

Tunable and multifunctional eukaryotic transcription factors based on CRISPR/Cas. Farzadfard F, Perli SD, Lu TK. ACS Synth Biol. 2013 Oct 18;2(10):604-13. doi: 10.1021/sb400081r. Epub 2013 Sep 11. http://www.ncbi.nlm.nih.gov/pubmed/23977949

Sequence of U6 promoter:

In vivo genome editing using Staphylococcus aureus Cas9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. Nature. 2015 Apr 9;520(7546):186-91. doi: 10.1038/nature14299. Epub 2015 Apr 1. http://www.ncbi.nlm.nih.gov/pubmed/25830891