Difference between revisions of "Part:BBa K2457004"

(Usage and Biology)
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Composed by the JK26 promoter, the coding sequence of the Green Fluorescent Protein (GFP) and the ilvBN terminator. It has 1810bp and was designed to contain two homologous arms with 300bp from the Operon Lac in the bacterial genome. Between the homologous arms, lies the GFP gene, flanked by NheI sites, allowing an RFC10 versatile one-step compatibility, the insertion of any other gene in GFP site. It has this structure especially for homology recognition by RecA enzyme which will break through the BioBrick to restore the cleaved sequence in the genome, having the aimed gene integrated.
 
Composed by the JK26 promoter, the coding sequence of the Green Fluorescent Protein (GFP) and the ilvBN terminator. It has 1810bp and was designed to contain two homologous arms with 300bp from the Operon Lac in the bacterial genome. Between the homologous arms, lies the GFP gene, flanked by NheI sites, allowing an RFC10 versatile one-step compatibility, the insertion of any other gene in GFP site. It has this structure especially for homology recognition by RecA enzyme which will break through the BioBrick to restore the cleaved sequence in the genome, having the aimed gene integrated.
 
<p>
 
<p>
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===Characterization===
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[[File:BBa K2457004 electropherogram.png]]
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Figure 1:Sequencing electropherogram from BBa_K2457004
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[[File:BBa K2457004 aligmnent.png]]
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Figure 2:Alignment of the designed sequence and our final construction from BBa_K2457004.
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<span class='h3bb'>Sequence and Features</span></p>
 
<span class='h3bb'>Sequence and Features</span></p>
 
<partinfo>BBa_K2457004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2457004 SequenceAndFeatures</partinfo>

Revision as of 07:54, 1 November 2017


Donor DNA for HDR into LacZ

This BioBrick is the key part of the CRISPeasy-in (CRIN) framework, developed in 2017 by the Amazonas_Brazil team to make easier and accessible the genome editing based on CRISPR/Cas9. In this construction stays the gene of the template DNA that will be inserted in the edited genome. It is the first registry vector for a Donor DNA.

Usage and Biology

Composed by the JK26 promoter, the coding sequence of the Green Fluorescent Protein (GFP) and the ilvBN terminator. It has 1810bp and was designed to contain two homologous arms with 300bp from the Operon Lac in the bacterial genome. Between the homologous arms, lies the GFP gene, flanked by NheI sites, allowing an RFC10 versatile one-step compatibility, the insertion of any other gene in GFP site. It has this structure especially for homology recognition by RecA enzyme which will break through the BioBrick to restore the cleaved sequence in the genome, having the aimed gene integrated.

Characterization

BBa K2457004 electropherogram.png Figure 1:Sequencing electropherogram from BBa_K2457004

BBa K2457004 aligmnent.png Figure 2:Alignment of the designed sequence and our final construction from BBa_K2457004.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 321
    Illegal NheI site found at 1164
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]