Difference between revisions of "Part:BBa K2457005"
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<partinfo>BBa_K2457005 short</partinfo>
 
<partinfo>BBa_K2457005 short</partinfo>
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<p>This BioBrick is composed by BBa_K2457000 (AraC + pBAD) and BBa_K2457001 (Cas9), where AraC + pBAD regules the Cas9 expression through your repressor and your inductor: Glucose and L-Arabinose, respectively, and Cas9 is a protein that cleaves a target sequence in the genome. This module was developed by the Amazonas_Brazil team to compose the CRISPeasy toolbox, subdivided in CRISPeasy-out and CRISPeasy-in.</p>
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<p>This BioBrick is composed by BBa_K2457000 (AraC + pBAD) and BBa_K2457001 (Cas9), where AraC + pBAD regulates the Cas9 expression through your repressor and your inductor: Glucose and L-Arabinose, respectively, and Cas9 is a protein that cleaves a target sequence in the genome. This module was developed by the Amazonas_Brazil team to compose the CRISPeasy toolbox, subdivided in CRISPeasy-out and CRISPeasy-in.</p>
 
<p><b>CRISPeasy-out</b> is a standardized knock out approach that aims to turn the current method in an easier way to engineer the bacterial genome by Non-Homologous End Joining (NHEJ). </p>
 
<p><b>CRISPeasy-out</b> is a standardized knock out approach that aims to turn the current method in an easier way to engineer the bacterial genome by Non-Homologous End Joining (NHEJ). </p>
 
<p><b>CRISPeasy-in</b> is a standardized knock-in approach that aims to turn the current method in an easier way to insert a donor template into the bacterial genome by Homology-Directed Repair (HDR).</p>
 
<p><b>CRISPeasy-in</b> is a standardized knock-in approach that aims to turn the current method in an easier way to insert a donor template into the bacterial genome by Homology-Directed Repair (HDR).</p>

Revision as of 01:03, 2 November 2017


Standardized pBAD_Cas9 device

This BioBrick is composed by BBa_K2457000 (AraC + pBAD) and BBa_K2457001 (Cas9), where AraC + pBAD regulates the Cas9 expression through your repressor and your inductor: Glucose and L-Arabinose, respectively, and Cas9 is a protein that cleaves a target sequence in the genome. This module was developed by the Amazonas_Brazil team to compose the CRISPeasy toolbox, subdivided in CRISPeasy-out and CRISPeasy-in.

CRISPeasy-out is a standardized knock out approach that aims to turn the current method in an easier way to engineer the bacterial genome by Non-Homologous End Joining (NHEJ).

CRISPeasy-in is a standardized knock-in approach that aims to turn the current method in an easier way to insert a donor template into the bacterial genome by Homology-Directed Repair (HDR).

BBa K2457005 circuit.png

Usage and Biology

Design

Figure 1: BBa K2457005 circuit

Characterization

Sequencing

BBa K2457005 electropherogram.png

Figure 2: Sequencing electropherogram from BBa_K2457005.

BBa K2457005 aligmnent.png

Figure 3: Alignment of the designed sequence and our final construction from BBa_K2457005.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1207
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1042
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1024