Difference between revisions of "Part:BBa K1699004:Design"

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1699004 short</partinfo>
 
<partinfo>BBa_K1699004 short</partinfo>
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===Design Notes===
 
===Design Notes===
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Cloned into pSBC13 using EcoRI and PstI restriction sites.
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Designed using Benchling to target human UBB gene and to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restriction sites.
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<br />F. tagtcatggcggccgcgtcgacG
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<br />R. actgacatgcggccgcttaattaaCTGC
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===Source===
 
===Source===
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Sequence of gRNA: designed using CRISPR tool in Benchling to target human UBB gene.
  
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gRNA for UBB by IDT de-novo syntesis.
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Sequence of U6 promoter:
  
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===References===
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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
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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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Latest revision as of 12:28, 13 September 2015

gRNA for SaCas9 targeting human ubiquitin B gene 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 target human UBB gene and to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restriction sites.

Source

Sequence of gRNA: designed using CRISPR tool in Benchling to target human UBB gene.

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