Difference between revisions of "Part:BBa K1773003:Design"
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<partinfo>BBa_K1773003 SequenceAndFeatures</partinfo>
 
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[[File:Cas3 1.JPG|300px|thumb|(Restriction with EcoRI (E), XbaI (X) and PstI (P) of Wild type (WT) Cas3 gene compared to mutated Cas3 genes. The first mutant plasmid has two successful mutations, whereas the second mutant plasmid only has one successfully mutated restriction site.)|left]]
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LALALALA
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===Design Notes===
 
===Design Notes===
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no
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Gene mutagenesis was performed using Invitrogen GeneArt® Site-Directed Mutagenesis PLUS System kit.
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These mutagenic primers were used:
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EcoRI mutagenesis: <br />
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F: cgttttcaatggcagaatccggcatttgatttggca<br />
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R: tgccaaatcaaatgccggattctgccattgaaaacg<br />
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XbaI Mutagenesis:<br />
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F: cgatcatcattattcttctctggatgctgatgttaatttggg<br />
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R: cccaaattaacatcagcatccagagaagaataatgatgatcg<br />
  
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PstI Mutagenesis:<br />
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F: gaagactgaactgcctgcggttaaacaacataaac<br />
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Re: gtttatgttgtttaaccgcaggcagttcagtcttc<br />
  
  
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===Source===
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Primers for adding Biobrick prefix and suffix:
  
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A.actinomycetemcomitans D7-S
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Fw: ttaaggaattcgcggccgcttctagatgaatatcttacttgttagccaatgccaaaaaaa
  
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===References===
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Rev: cggctgcagcggccgctactagtagagatcttcctgccaaaaacccagcgttt

Latest revision as of 15:11, 10 September 2015

I-F type CRISPR-Cas Cas3 gene


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2623
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1344
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2307
    Illegal AgeI site found at 2680
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Gene mutagenesis was performed using Invitrogen GeneArt® Site-Directed Mutagenesis PLUS System kit.

These mutagenic primers were used:

EcoRI mutagenesis:
F: cgttttcaatggcagaatccggcatttgatttggca
R: tgccaaatcaaatgccggattctgccattgaaaacg

XbaI Mutagenesis:
F: cgatcatcattattcttctctggatgctgatgttaatttggg
R: cccaaattaacatcagcatccagagaagaataatgatgatcg

PstI Mutagenesis:
F: gaagactgaactgcctgcggttaaacaacataaac
Re: gtttatgttgtttaaccgcaggcagttcagtcttc


Primers for adding Biobrick prefix and suffix:

Fw: ttaaggaattcgcggccgcttctagatgaatatcttacttgttagccaatgccaaaaaaa

Rev: cggctgcagcggccgctactagtagagatcttcctgccaaaaacccagcgttt