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A plasmid toolset for CRISPR-mediated genome editing and CRISPRi gene regulation in Escherichia coli.
Jervis AJ, Hanko EKR, Dunstan MS, Robinson CJ, Takano E, Scrutton NS
Microb Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
153034pSIMcpf1For CRISPR-Cas12a genome editing in E. coli. Contains; constituitively expressed Ascpf1 gene, a heat-inducible lambda red recombination system, an arabinose-inducible CRISPR array targeting pMB1.
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153035pTF-ahpC-rfpFor CRISPR-Cas12a genome editing in E. coli. Contains a constituitively expressed CRISPR array targeting the ahpC gene in E. coli and donor DNA to insert an rfp translational fusion in the ahpC gene.
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153036pTF-lacZFor CRISPR-Cas12a genome editing in E. coli for the insertion of large DNA fragments at the lacZ locus. Contains a constitutively expressed CRISPR array targeting the lacZ gene . No cargo DNA.
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153037pTF-lacZ-rfpCRISPR-Cas12a genome editing in E. coli. Contains a constitutively CRISPR array targeting the lacZ gene in E. coli genome and donor DNA to insert an constitutively expressed rfp reporter gene.
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153038pBbS8c-ddcpf1-rfpFor CRISPRi. Arabinose-inducible ddcpf1 gene with CRISPR array targeting the mrfp reporter gene.
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153039pBbS8c-ddcpf1-ΔFor CRISPRi. Arabinose-inducible ddcpf1 gene with CRISPR array lacking a gene-specific spacer.
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