Difference between revisions of "Part:BBa K3738021"

 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
<partinfo>BBa_K3738019 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3738021 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
<partinfo>BBa_K3738019 parameters</partinfo>
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<partinfo>BBa_K3738021 parameters</partinfo>
 
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Latest revision as of 17:04, 20 October 2021


Lbu-Cas13a with an N-terminal Anionic Tag and C-Terminal 6XHistidine Tag

Cas13a is an enzyme originating from Leptotrichia buccalis (Lbu) which functions to cleave single-stranded RNAs (ssRNAs); particularly mRNAs. This function is achieved following protein-RNA complex formation with CRISPR RNA (crRNA) via crRNA backbone contacts with residues from the Helical-2, HEPN1, and Linker domains of Cas13a. The crRNA contains a spacer region coding for a direct repeat stem loop as well as a region complementary to target ssRNAs. Once the enzyme complex interacts with a target ssRNA, a structural conformation change occurs within the domains of the protein that permits active site formation for non-discriminate ssRNA cleavage (O’Connel et al., 2019).

O'Connell MR. Molecular Mechanisms of RNA Targeting by Cas13-containing Type VI CRISPR–Cas Systems. Journal of Molecular Biology. 2019;431(1):66-87. doi: https://doi.org/10.1016/j.jmb.2018.06.029.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1117
    Illegal PstI site found at 1972
    Illegal PstI site found at 2911
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1117
    Illegal PstI site found at 1972
    Illegal PstI site found at 2911
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 398
    Illegal BglII site found at 1334
    Illegal BglII site found at 1544
    Illegal BglII site found at 1598
    Illegal BglII site found at 1829
    Illegal BglII site found at 2102
    Illegal BglII site found at 2588
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1117
    Illegal PstI site found at 1972
    Illegal PstI site found at 2911
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1117
    Illegal PstI site found at 1972
    Illegal PstI site found at 2911
    Illegal NgoMIV site found at 2050
    Illegal NgoMIV site found at 2686
  • 1000
    COMPATIBLE WITH RFC[1000]