{"entryType":"UniProtKB reviewed (Swiss-Prot)","primaryAccession":"Q46896","secondaryAccessions":["Q2MA74"],"uniProtkbId":"CAS1_ECOLI","entryAudit":{"firstPublicDate":"2001-06-01","lastAnnotationUpdateDate":"2026-09-02","lastSequenceUpdateDate":"1996-11-01","entryVersion":154,"sequenceVersion":1},"annotationScore":5.0,"organism":{"scientificName":"Escherichia coli (strain K12)","taxonId":83333,"lineage":["Bacteria","Pseudomonadati","Pseudomonadota","Gammaproteobacteria","Enterobacterales","Enterobacteriaceae","Escherichia"]},"proteinExistence":"1: Evidence at protein level","proteinDescription":{"recommendedName":{"fullName":{"value":"CRISPR-associated endonuclease Cas1"},"ecNumbers":[{"value":"3.1.-.-"}]}},"genes":[{"geneName":{"value":"ygbT"},"synonyms":[{"value":"cas1"}],"orderedLocusNames":[{"value":"b2755"},{"value":"JW2725"}]}],"comments":[{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21255106"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24920831"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26284603"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"value":"CRISPR (clustered regularly interspaced short palindromic repeat), is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids) (PubMed:21255106, PubMed:24793649, PubMed:24920831). CRISPR clusters contain sequences complementary to antecedent mobile elements and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). The Cas1-Cas2 complex is involved in CRISPR adaptation, the first stage of CRISPR immunity, being required for the addition/removal of CRISPR spacers at the leader end of the CRISPR locus (PubMed:24793649, PubMed:24920831, PubMed:25707795). The Cas1-Cas2 complex introduces staggered nicks into both strands of the CRISPR array near the leader repeat and joins the 5'-ends of the repeat strands with the 3'-ends of the new spacer sequence (PubMed:24920831). Spacer DNA integration requires supercoiled target DNA and 3'-OH ends on the inserted (spacer) DNA and probably initiates with a nucleophilic attack of the C 3'-OH end of the protospacer on the minus strand of the first repeat sequence (PubMed:25707795). Expression of Cas1-Cas2 in a strain lacking both genes permits spacer acquisition (PubMed:24793649, PubMed:24920831). Non-specifically binds DNA; the Cas1-Cas2 complex preferentially binds CRISPR-locus DNA (PubMed:24793649). Highest binding is seen to a dual forked DNA complex with 3'-overhangs and a protospacer-adjacent motif-complement specifically positioned (PubMed:26478180). The protospacer DNA lies across a flat surface extending from 1 Cas1 dimer, across the Cas2 dimer and contacting the other Cas1 dimer; the 23 bp-long ds section of the DNA is bracketed by 1 Tyr-22 from each of the Cas1 dimers (PubMed:26478180, PubMed:26503043). Cas1 cuts within the 3'-overhang, to generate a 33-nucleotide DNA that is probably incorporated into the CRISPR leader by a cut-and-paste mechanism (PubMed:26478180). Cas1 alone endonucleolytically cleaves linear ssRNA, ssDNA and short (34 base) dsDNA as well as branched DNA substrates such as Holliday junctions, replication forks and 5'-flap DNA substrates (PubMed:21219465). In vitro catalyzes a concerted transesterification reaction on branched DNA, as would be expected during integration of protospacers into the CRISPR leader sequence; Cas2 is not required in vitro. This reaction requires a 3'-OH group at the branch point (PubMed:26284603). Genetic interactions suggest Cas1 interacts with components of the RecBC and RuvB DNA repair systems (PubMed:21219465)"}],"commentType":"FUNCTION"},{"commentType":"COFACTOR","cofactors":[{"name":"Mg(2+)","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26284603"}],"cofactorCrossReference":{"database":"ChEBI","id":"CHEBI:18420"}}],"note":{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26284603"}],"value":"Protospacer integration in vitro also occurs with Mn(2+) and also requires low concentrations of KCl (PubMed:25707795). The transesterification function works equally well with Mg(2+), Mn(2+) or Co(2+) in vitro (PubMed:26284603)."}]}},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"}],"value":"Nuclease activity partially inhibited by CasE (PubMed:21219465)"}],"commentType":"ACTIVITY REGULATION"},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24920831"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"},{"evidenceCode":"ECO:0000305","source":"Reference","id":"Ref.11"}],"value":"Homodimer (PubMed:21219465). Part of the Cas1-Cas2 complex (PubMed:24793649, PubMed:24920831, PubMed:25707795, PubMed:26478180, PubMed:26503043, Ref.11). Interacts with RecB, RecC, RuvB, CasC and CasE (PubMed:21219465). Forms a hexamer with 2 Cas1 dimers sandwiching a Cas2 dimer (PubMed:24793649, PubMed:26478180). The DNA lies across a flat surface extending from 1 Cas1 dimer, across the Cas2 dimer and contacting the other Cas1 dimer. Only 1 Cas1 protein from each dimer is catalytic, the other interacts with the Cas2 dimer and possibly target DNA (PubMed:26478180, PubMed:26503043)"}],"commentType":"SUBUNIT"},{"commentType":"INTERACTION","interactions":[{"interactantOne":{"uniProtKBAccession":"Q46896","intActId":"EBI-1130209"},"interactantTwo":{"uniProtKBAccession":"P76339","geneName":"hprS","intActId":"EBI-554869"},"numberOfExperiments":3,"organismDiffer":false},{"interactantOne":{"uniProtKBAccession":"Q46896","intActId":"EBI-1130209"},"interactantTwo":{"uniProtKBAccession":"P45956","geneName":"ygbF","intActId":"EBI-9150552"},"numberOfExperiments":8,"organismDiffer":false},{"interactantOne":{"uniProtKBAccession":"Q46896","intActId":"EBI-1130209"},"interactantTwo":{"uniProtKBAccession":"Q46896","geneName":"ygbT","intActId":"EBI-1130209"},"numberOfExperiments":4,"organismDiffer":false}]},{"commentType":"SUBCELLULAR LOCATION","note":{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"}],"value":"In 15% of cell localizes to discrete nucleoid foci (probable DNA damage sites) upon treatment with mitomycin C (MMC) for 2 hours (PubMed:21219465)"}]},"subcellularLocations":[{"location":{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"}],"value":"Cytoplasm","id":"SL-0086"}}]},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"19429622"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"20132443"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21255106"}],"value":"Repressed by H-NS (PubMed:20132443). Activated by LeuO (PubMed:19429622). Activated by the BaeSR two-component regulatory system, possibly due to envelope stress (PubMed:21255106). Part of the casABCDE-ygbT-ygbF operon (PubMed:19429622)"}],"commentType":"INDUCTION"},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"value":"Substrate DNA-binding induces large structural changes that generate a surface for DNA-binding across the Cas2 dimer and formation of an optimal catalytic site (PubMed:26478180)"}],"commentType":"DOMAIN"},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21255106"}],"value":"Not essential. Increased sensitivity to MMC and UV light; double ygbT-ruvA, ruvB or ruvC disruptions have no further phenotype suggesting Cas1 functions in the same DNA repair pathway (PubMed:21219465). Function in DNA repair also seems to require CRISPRs (PubMed:21219465). Cells elongate after 2 hours growth in MMC; they are even longer in double ygbT-ruvA, ruvB or ruvC disruptions, suggesting Cas1 may also function in chromosome segregation (PubMed:21219465). Loss of plasmid silencing (PubMed:21255106)"}],"commentType":"DISRUPTION PHENOTYPE"},{"texts":[{"evidences":[{"evidenceCode":"ECO:0000305"}],"value":"Belongs to the CRISPR-associated endonuclease Cas1 family"}],"commentType":"SIMILARITY"}],"features":[{"type":"Chain","location":{"start":{"value":1,"modifier":"EXACT"},"end":{"value":305,"modifier":"EXACT"}},"description":"CRISPR-associated endonuclease Cas1","featureId":"PRO_0000169315"},{"type":"Region","location":{"start":{"value":96,"modifier":"EXACT"},"end":{"value":278,"modifier":"EXACT"}},"description":"Sufficient for cleavage of ssRNA, ssDNA and Holliday junction DNA"},{"type":"Region","location":{"start":{"value":278,"modifier":"EXACT"},"end":{"value":305,"modifier":"EXACT"}},"description":"Disordered","evidences":[{"evidenceCode":"ECO:0000256","source":"SAM","id":"MobiDB-lite"}]},{"type":"Binding site","location":{"start":{"value":141,"modifier":"EXACT"},"end":{"value":141,"modifier":"EXACT"}},"description":"","featureCrossReferences":[{"database":"ChEBI","id":"CHEBI:18420"}],"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"},{"evidenceCode":"ECO:0007744","source":"PDB","id":"5DS5"}],"ligand":{"name":"Mg(2+)","id":"ChEBI:CHEBI:18420"}},{"type":"Binding site","location":{"start":{"value":208,"modifier":"EXACT"},"end":{"value":208,"modifier":"EXACT"}},"description":"","featureCrossReferences":[{"database":"ChEBI","id":"CHEBI:18420"}],"evidences":[{"evidenceCode":"ECO:0000305","source":"PubMed","id":"26503043"}],"ligand":{"name":"Mg(2+)","id":"ChEBI:CHEBI:18420"}},{"type":"Binding site","location":{"start":{"value":221,"modifier":"EXACT"},"end":{"value":221,"modifier":"EXACT"}},"description":"","featureCrossReferences":[{"database":"ChEBI","id":"CHEBI:18420"}],"evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"},{"evidenceCode":"ECO:0007744","source":"PDB","id":"5DS5"}],"ligand":{"name":"Mg(2+)","id":"ChEBI:CHEBI:18420"}},{"type":"Mutagenesis","location":{"start":{"value":22,"modifier":"EXACT"},"end":{"value":22,"modifier":"EXACT"}},"description":"Slightly decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"Y","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":22,"modifier":"EXACT"},"end":{"value":22,"modifier":"EXACT"}},"description":"Nearly wild-type spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"Y","alternativeSequences":["F"]}},{"type":"Mutagenesis","location":{"start":{"value":41,"modifier":"EXACT"},"end":{"value":41,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["E"]}},{"type":"Mutagenesis","location":{"start":{"value":59,"modifier":"EXACT"},"end":{"value":59,"modifier":"EXACT"}},"description":"Loss of spacer acquisition in vivo, decreased protospacer binding.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":59,"modifier":"EXACT"},"end":{"value":59,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vitro, 250-fold decreased affinity for protospacer DNA.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["D"]}},{"type":"Mutagenesis","location":{"start":{"value":66,"modifier":"EXACT"},"end":{"value":66,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vitro, 250-fold decreased affinity for protospacer DNA.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["D"]}},{"type":"Mutagenesis","location":{"start":{"value":66,"modifier":"EXACT"},"end":{"value":66,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["E"]}},{"type":"Mutagenesis","location":{"start":{"value":84,"modifier":"EXACT"},"end":{"value":84,"modifier":"EXACT"}},"description":"Decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":84,"modifier":"EXACT"},"end":{"value":84,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["E"]}},{"type":"Mutagenesis","location":{"start":{"value":141,"modifier":"EXACT"},"end":{"value":141,"modifier":"EXACT"}},"description":"No cleavage of any substrates, no restoration of UV or mitomycin C (MMC) resistance (PubMed:21219465). Loss of spacer acquisition in vivo (PubMed:24793649).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"}],"alternativeSequence":{"originalSequence":"E","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":149,"modifier":"EXACT"},"end":{"value":149,"modifier":"EXACT"}},"description":"No effect on in vitro protospacer integration.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"}],"alternativeSequence":{"originalSequence":"Y","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":165,"modifier":"EXACT"},"end":{"value":165,"modifier":"EXACT"}},"description":"No effect on in vitro protospacer integration (PubMed:25707795). Alone significantly decreased protospacer acquisition in vivo (PubMed:26478180). Loss of protospacer acquisition, decreased protospacer binding; in association with A-170, significantly decreased protospacer binding; in association with A-217 (PubMed:26478180).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"Y","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":170,"modifier":"EXACT"},"end":{"value":170,"modifier":"EXACT"}},"description":"Alone significantly decreased protospacer acquisition in vivo (PubMed:26478180). Decreased protospacer binding; in association with A-170 (PubMed:26478180).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"W","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":184,"modifier":"EXACT"},"end":{"value":184,"modifier":"EXACT"}},"description":"No cleavage of any substrates.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"}],"alternativeSequence":{"originalSequence":"T","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":188,"modifier":"EXACT"},"end":{"value":188,"modifier":"EXACT"}},"description":"Partial inhibition of cleavage (PubMed:21219465). No effect on in vitro protospacer integration (PubMed:25707795). Significantly decreased protospacer acquisition in vivo (PubMed:26478180).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"Y","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":208,"modifier":"EXACT"},"end":{"value":208,"modifier":"EXACT"}},"description":"No cleavage of any substrates, no restoration of UV or MMC resistance (PubMed:21219465). Loss of spacer acquisition in vivo (PubMed:24793649, PubMed:25707795, PubMed:26478180).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"H","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":211,"modifier":"EXACT"},"end":{"value":211,"modifier":"EXACT"}},"description":"No cleavage of any substrates.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"}],"alternativeSequence":{"originalSequence":"K","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":217,"modifier":"EXACT"},"end":{"value":217,"modifier":"EXACT"}},"description":"No effect on in vitro protospacer integration (PubMed:25707795). Alone significantly decreased protospacer acquisition in vivo (PubMed:26478180). Significantly decreased protospacer binding; in association with A-165 (PubMed:26478180).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"Y","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":218,"modifier":"EXACT"},"end":{"value":218,"modifier":"EXACT"}},"description":"No cleavage of any substrates, no restoration of UV or MMC resistance (PubMed:21219465). Loss of spacer acquisition in vivo (PubMed:24793649).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"}],"alternativeSequence":{"originalSequence":"D","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":221,"modifier":"EXACT"},"end":{"value":221,"modifier":"EXACT"}},"description":"No cleavage of any substrates (PubMed:21219465). Loss of spacer acquisition in vivo (PubMed:24793649, PubMed:24920831, PubMed:25707795). No cleavage of CRISPR leader in preparation for spacer integration (PubMed:24920831).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24920831"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"}],"alternativeSequence":{"originalSequence":"D","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":224,"modifier":"EXACT"},"end":{"value":224,"modifier":"EXACT"}},"description":"No cleavage of any substrates (PubMed:21219465). Loss of spacer acquisition in vivo (PubMed:24793649, PubMed:25707795).","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"21219465"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"},{"evidenceCode":"ECO:0000269","source":"PubMed","id":"25707795"}],"alternativeSequence":{"originalSequence":"K","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":245,"modifier":"EXACT"},"end":{"value":248,"modifier":"EXACT"}},"description":"Loss of spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26478180"}],"alternativeSequence":{"originalSequence":"REVR","alternativeSequences":["AEVA"]}},{"type":"Mutagenesis","location":{"start":{"value":245,"modifier":"EXACT"},"end":{"value":245,"modifier":"EXACT"}},"description":"No effect on spacer acquisition.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":245,"modifier":"EXACT"},"end":{"value":245,"modifier":"EXACT"}},"description":"Decreased spacer acquisition.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["D"]}},{"type":"Mutagenesis","location":{"start":{"value":245,"modifier":"EXACT"},"end":{"value":245,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["E"]}},{"type":"Mutagenesis","location":{"start":{"value":248,"modifier":"EXACT"},"end":{"value":248,"modifier":"EXACT"}},"description":"Dramatically decreased spacer acquisition in vivo.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"26503043"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["E"]}},{"type":"Mutagenesis","location":{"start":{"value":252,"modifier":"EXACT"},"end":{"value":252,"modifier":"EXACT"}},"description":"No effect on spacer acquisition.","evidences":[{"evidenceCode":"ECO:0000269","source":"PubMed","id":"24793649"}],"alternativeSequence":{"originalSequence":"R","alternativeSequences":["A"]}},{"type":"Mutagenesis","location":{"start":{"value":252,"modifier":"EXACT"},"end":{"value":252,"modifier":"EXACT"}},"description":"Loss of spacer acquisition, no Cas1-Cas2 complex formation, loss of CRISPRDNA-binding by complex. 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