Difference between revisions of "Part:BBa K1773005"

 
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<partinfo>BBa_K1773005 short</partinfo>
 
<partinfo>BBa_K1773005 short</partinfo>
  
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I-F type CRISPR-Cas Cascade complex mutated for biobrick assembly.  
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I-F type CRISPR-Cas Cascade complex, which has a mutated EcoRI restriction site to make it biobrick compatible. This protein complex needs to be coexpressed with a crRNA region, to make a functional Cascade ribonucleoprotein complex. Together With Cas3 (BBa_K1773003) degrades DNA.
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===Usage and Biology===
 
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1773005 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1773005 SequenceAndFeatures</partinfo>
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===Usage and Biology===
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Cascade (CRISPR Associated Complex for Antiviral Defence) complex is an essential component of I-type CRISPR-Cas systems. This ribonucleoprotein complex is coded by 4 genes in an operon : Csy1, Csy2, Csy3 and Cas6f, with stoichiometry of 1:1:6:1 respectively. Also it has a crRNA molecule, witch acts like a guide, that brings the whole complex to the target dsDNA sequence. Each Cascade complex has a unique Protospacer adjacent motive (PAM) recognition site. The recognisable PAM sequence of this specific I-F Cascade complex from  Aggregatibacter actinomycetemcomitans recognises CC nucleotides directly upstream of the protospacer (the target sequence). In other words, a target sequence needs to have a CC pair on the non-complementary strand of DNA -2 -1 positions of the protospacer. When Cascade complex finds and recognises the DNA target, then Cas3 (BBa_K1773003) is recruited for DNA degradation.
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The wild type Cascade complex had EcoRI restriction site inside the coding region of Csy1 gene.
  
  

Revision as of 14:20, 10 September 2015

I-F type CRISPR-Cas Cascade complex

I-F type CRISPR-Cas Cascade complex, which has a mutated EcoRI restriction site to make it biobrick compatible. This protein complex needs to be coexpressed with a crRNA region, to make a functional Cascade ribonucleoprotein complex. Together With Cas3 (BBa_K1773003) degrades DNA.



Sequence and Features


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 NgoMIV site found at 1790
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2307

Usage and Biology

Cascade (CRISPR Associated Complex for Antiviral Defence) complex is an essential component of I-type CRISPR-Cas systems. This ribonucleoprotein complex is coded by 4 genes in an operon : Csy1, Csy2, Csy3 and Cas6f, with stoichiometry of 1:1:6:1 respectively. Also it has a crRNA molecule, witch acts like a guide, that brings the whole complex to the target dsDNA sequence. Each Cascade complex has a unique Protospacer adjacent motive (PAM) recognition site. The recognisable PAM sequence of this specific I-F Cascade complex from Aggregatibacter actinomycetemcomitans recognises CC nucleotides directly upstream of the protospacer (the target sequence). In other words, a target sequence needs to have a CC pair on the non-complementary strand of DNA -2 -1 positions of the protospacer. When Cascade complex finds and recognises the DNA target, then Cas3 (BBa_K1773003) is recruited for DNA degradation.

The wild type Cascade complex had EcoRI restriction site inside the coding region of Csy1 gene.