Difference between revisions of "Part:BBa K2457006"
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2457006 short</partinfo>
 
<partinfo>BBa_K2457006 short</partinfo>
 +
<p> This BioBrick is composed by BBa_K2457000 (AraC + pBAD), BBa_K2457001 (Cas9) and BBa_K2457002 (simple guide RNA). Which AraC + pBAD regules the Cas9 expression through your repressor and your inductor: Glucose and L-Arabinose, respectively. Cas9 is a protein that with the assistance of sgRNA cleaves a target-site in the bacterial genome. This construction was developed by Amazonas_Brazil team to be a fundamental part of the CRISPeasy toolbox, divided in CRISPReasy-out and CRISPReasy-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-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>
  
−
descrever
 
  
 
[[File:BBa K2457006 circuit.png]]
 
[[File:BBa K2457006 circuit.png]]

Revision as of 01:02, 2 November 2017


Cas9 device + Optimized sgRNA

This BioBrick is composed by BBa_K2457000 (AraC + pBAD), BBa_K2457001 (Cas9) and BBa_K2457002 (simple guide RNA). Which AraC + pBAD regules the Cas9 expression through your repressor and your inductor: Glucose and L-Arabinose, respectively. Cas9 is a protein that with the assistance of sgRNA cleaves a target-site in the bacterial genome. This construction was developed by Amazonas_Brazil team to be a fundamental part of the CRISPeasy toolbox, divided in CRISPReasy-out and CRISPReasy-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 K2457006 circuit.png

Figure 1: BBa K2457006 circuit

Usage and Biology

Design

Characterization

Sequencing

BBa K2457006 electropherogram.png

Figure 2:Sequencing electropherogram from BBa_K2457006.

BBa K2457006 alignment.png

Figure 3:Alignment of the designed sequence and our final construction from BBa_K2457006.

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