Part:BBa_K2457005
Standardized pBAD_Cas9 device
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.
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).
Usage and Biology
Design
Figure 1: BBa K2457005 circuit
Characterization
Sequencing
Figure 2: Sequencing electropherogram from BBa_K2457005.
Figure 3: Alignment of the designed sequence and our final construction from BBa_K2457005.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1207
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1042
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1024
| None |