CAS9_STRP1
ID CAS9_STRP1 Reviewed; 1368 AA.
AC Q99ZW2;
DT 11-JUL-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2001, sequence version 1.
DT 03-AUG-2022, entry version 129.
DE RecName: Full=CRISPR-associated endonuclease Cas9/Csn1 {ECO:0000255|HAMAP-Rule:MF_01480};
DE EC=3.1.-.- {ECO:0000255|HAMAP-Rule:MF_01480};
DE AltName: Full=SpCas9 {ECO:0000303|PubMed:23287718};
DE AltName: Full=SpyCas9 {ECO:0000303|PubMed:24505130};
GN Name=cas9 {ECO:0000255|HAMAP-Rule:MF_01480, ECO:0000303|PubMed:22745249};
GN Synonyms=csn1 {ECO:0000303|PubMed:21455174}; OrderedLocusNames=SPy_1046;
OS Streptococcus pyogenes serotype M1.
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=301447;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=11296296; DOI=10.1073/pnas.071559398;
RA Ferretti J.J., McShan W.M., Ajdic D.J., Savic D.J., Savic G., Lyon K.,
RA Primeaux C., Sezate S., Suvorov A.N., Kenton S., Lai H.S., Lin S.P.,
RA Qian Y., Jia H.G., Najar F.Z., Ren Q., Zhu H., Song L., White J., Yuan X.,
RA Clifton S.W., Roe B.A., McLaughlin R.E.;
RT "Complete genome sequence of an M1 strain of Streptococcus pyogenes.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:4658-4663(2001).
RN [2]
RP FUNCTION IN CRISPR-MEDIATED PLASMID DEFENSE, AND DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=21455174; DOI=10.1038/nature09886;
RA Deltcheva E., Chylinski K., Sharma C.M., Gonzales K., Chao Y.,
RA Pirzada Z.A., Eckert M.R., Vogel J., Charpentier E.;
RT "CRISPR RNA maturation by trans-encoded small RNA and host factor RNase
RT III.";
RL Nature 471:602-607(2011).
RN [3]
RP FUNCTION AS A DNA ENDONUCLEASE, FUNCTION AS AN EXONUCLEASE, COFACTOR,
RP ACTIVITY REGULATION, DNA-BINDING, POSSIBLE BIOTECHNOLOGY, AND MUTAGENESIS
RP OF ASP-10 AND HIS-840.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=22745249; DOI=10.1126/science.1225829;
RA Jinek M., Chylinski K., Fonfara I., Hauer M., Doudna J.A., Charpentier E.;
RT "A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial
RT immunity.";
RL Science 337:816-821(2012).
RN [4]
RP BIOTECHNOLOGY IN HUMAN AND MOUSE CELLS.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=23287718; DOI=10.1126/science.1231143;
RA Cong L., Ran F.A., Cox D., Lin S., Barretto R., Habib N., Hsu P.D., Wu X.,
RA Jiang W., Marraffini L.A., Zhang F.;
RT "Multiplex genome engineering using CRISPR/Cas systems.";
RL Science 339:819-823(2013).
RN [5]
RP BIOTECHNOLOGY IN HUMAN CELLS.
RX PubMed=23287722; DOI=10.1126/science.1232033;
RA Mali P., Yang L., Esvelt K.M., Aach J., Guell M., Dicarlo J.E.,
RA Norville J.E., Church G.M.;
RT "RNA-guided human genome engineering via Cas9.";
RL Science 339:823-826(2013).
RN [6]
RP BIOTECHNOLOGY IN ZEBRAFISH EMBRYOS.
RX PubMed=23360964; DOI=10.1038/nbt.2501;
RA Hwang W.Y., Fu Y., Reyon D., Maeder M.L., Tsai S.Q., Sander J.D.,
RA Peterson R.T., Yeh J.R., Joung J.K.;
RT "Efficient genome editing in zebrafish using a CRISPR-Cas system.";
RL Nat. Biotechnol. 31:227-229(2013).
RN [7]
RP BIOTECHNOLOGY IN BACTERIA, AND MUTAGENESIS OF GLY-1132.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=23360965; DOI=10.1038/nbt.2508;
RA Jiang W., Bikard D., Cox D., Zhang F., Marraffini L.A.;
RT "RNA-guided editing of bacterial genomes using CRISPR-Cas systems.";
RL Nat. Biotechnol. 31:233-239(2013).
RN [8]
RP BIOTECHNOLOGY IN HUMAN CELLS.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=23360966; DOI=10.1038/nbt.2507;
RA Cho S.W., Kim S., Kim J.M., Kim J.S.;
RT "Targeted genome engineering in human cells with the Cas9 RNA-guided
RT endonuclease.";
RL Nat. Biotechnol. 31:230-232(2013).
RN [9]
RP BIOTECHNOLOGY IN HUMAN CELLS.
RX PubMed=23386978; DOI=10.7554/elife.00471.009;
RA Jinek M., East A., Cheng A., Lin S., Ma E., Doudna J.;
RT "RNA-programmed genome editing in human cells.";
RL Elife 2:E00471-E00471(2013).
RN [10]
RP FUNCTION, ACTIVITY REGULATION, AND IMPORTANCE OF PAM SEQUENCES.
RX PubMed=24476820; DOI=10.1038/nature13011;
RA Sternberg S.H., Redding S., Jinek M., Greene E.C., Doudna J.A.;
RT "DNA interrogation by the CRISPR RNA-guided endonuclease Cas9.";
RL Nature 507:62-67(2014).
RN [11]
RP FUNCTION IN CRISPR-MEDIATED PLASMID DEFENSE, FUNCTION AS AN ENDONUCLEASE,
RP AND MUTAGENESIS OF ASP-10; GLU-762; HIS-840; ASN-854; ASN-863;
RP 982-HIS-HIS-983 AND ASP-986.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=24270795; DOI=10.1093/nar/gkt1074;
RA Fonfara I., Le Rhun A., Chylinski K., Makarova K.S., Lecrivain A.L.,
RA Bzdrenga J., Koonin E.V., Charpentier E.;
RT "Phylogeny of Cas9 determines functional exchangeability of dual-RNA and
RT Cas9 among orthologous type II CRISPR-Cas systems.";
RL Nucleic Acids Res. 42:2577-2590(2014).
RN [12]
RP RE-ENGINEERING, AND BIOTECHNOLOGY.
RX PubMed=26098369; DOI=10.1038/nature14592;
RA Kleinstiver B.P., Prew M.S., Tsai S.Q., Topkar V.V., Nguyen N.T., Zheng Z.,
RA Gonzales A.P., Li Z., Peterson R.T., Yeh J.R., Aryee M.J., Joung J.K.;
RT "Engineered CRISPR-Cas9 nucleases with altered PAM specificities.";
RL Nature 523:481-485(2015).
RN [13]
RP X-RAY CRYSTALLOGRAPHY (2.62 ANGSTROMS) IN COMPLEX WITH MANGANESE, STRUCTURE
RP BY ELECTRON MICROSCOPY IN COMPLEX WITH SGRNA AND TARGET DNA OR SGRNA ALONE,
RP FUNCTION, COFACTOR, ACTIVITY REGULATION, SUBUNIT, DOMAIN, DNA-BINDING, AND
RP MUTAGENESIS OF 475-PRO--ASN-477 AND 1125-ASP--ASP-1127.
RC STRAIN=ATCC 700294 / SF370 / Serotype M1;
RX PubMed=24505130; DOI=10.1126/science.1247997;
RA Jinek M., Jiang F., Taylor D.W., Sternberg S.H., Kaya E., Ma E., Anders C.,
RA Hauer M., Zhou K., Lin S., Kaplan M., Iavarone A.T., Charpentier E.,
RA Nogales E., Doudna J.A.;
RT "Structures of Cas9 endonucleases reveal RNA-mediated conformational
RT activation.";
RL Science 343:1247997-1247997(2014).
RN [14]
RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH SSGRNA AND
RP SINGLE-STRAND TARGET DNA, FUNCTION, SUBUNIT, DOMAIN, DNA-BINDING,
RP RNA-BINDING, AND MUTAGENESIS OF ASP-10; SER-15; ARG-66; ARG-70; ARG-74;
RP ARG-78; 97-PHE--ASP-150; ARG-165; 175-ASN--ARG-307; 312-ILE--SER-409;
RP GLU-762; HIS-840; ASN-854; ASN-863; HIS-982; HIS-983; ASP-986 AND
RP 1099-GLU--ASP-1368.
RX PubMed=24529477; DOI=10.1016/j.cell.2014.02.001;
RA Nishimasu H., Ran F.A., Hsu P.D., Konermann S., Shehata S.I., Dohmae N.,
RA Ishitani R., Zhang F., Nureki O.;
RT "Crystal structure of Cas9 in complex with guide RNA and target DNA.";
RL Cell 156:935-949(2014).
RN [15]
RP X-RAY CRYSTALLOGRAPHY (2.37 ANGSTROMS) IN COMPLEX WITH MAGNESIUM AND WITH
RP SGRNAS AND SINGLE-STRAND TARGET DNA, MUTAGENESIS OF HIS-840;
RP 1333-ARG--ARG-1335; ARG-1333 AND ARG-1335, DNA-BINDING, AND RNA-BINDING.
RX PubMed=25079318; DOI=10.1038/nature13579;
RA Anders C., Niewoehner O., Duerst A., Jinek M.;
RT "Structural basis of PAM-dependent target DNA recognition by the Cas9
RT endonuclease.";
RL Nature 513:569-573(2014).
RN [16]
RP X-RAY CRYSTALLOGRAPHY (2.90 ANGSTROMS) OF ACTIVE OR INACTIVE ENZYME IN
RP COMPLEX WITH SGRNA, DOMAIN, AND RNA-BINDING.
RX PubMed=26113724; DOI=10.1126/science.aab1452;
RA Jiang F., Zhou K., Ma L., Gressel S., Doudna J.A.;
RT "A Cas9-guide RNA complex preorganized for target DNA recognition.";
RL Science 348:1477-1481(2015).
RN [17]
RP X-RAY CRYSTALLOGRAPHY (3.40 ANGSTROMS) IN COMPLEX WITH SGRNA AND
RP DOUBLE-STRAND TARGET DNA, STRUCTURE BY ELECTRON MICROSCOPY IN COMPLEX WITH
RP SGRNA WITH AND WITHOUT DOUBLE-STRAND TARGET DNA, ACTIVITY REGULATION, AND
RP DOMAIN.
RX PubMed=26841432; DOI=10.1126/science.aad8282;
RA Jiang F., Taylor D.W., Chen J.S., Kornfeld J.E., Zhou K., Thompson A.J.,
RA Nogales E., Doudna J.A.;
RT "Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage.";
RL Science 351:867-871(2016).
CC -!- FUNCTION: CRISPR (clustered regularly interspaced short palindromic
CC repeat) is an adaptive immune system that provides protection against
CC mobile genetic elements (viruses, transposable elements and conjugative
CC plasmids) (PubMed:21455174). CRISPR clusters contain spacers, sequences
CC complementary to antecedent mobile elements, and target invading
CC nucleic acids. CRISPR clusters are transcribed and processed into
CC CRISPR RNA (crRNA). In type II CRISPR systems correct processing of
CC pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous
CC ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide
CC for ribonuclease 3-aided processing of pre-crRNA; Cas9 only stabilizes
CC the pre-crRNA:tracrRNA interaction and has no catalytic function in RNA
CC processing (PubMed:24270795). Subsequently Cas9/crRNA/tracrRNA
CC endonucleolytically cleaves linear or circular dsDNA target
CC complementary to the spacer; Cas9 is inactive in the absence of the 2
CC guide RNAs (gRNA). The target strand not complementary to crRNA is
CC first cut endonucleolytically, then trimmed 3'-5' exonucleolytically.
CC DNA-binding requires protein and both gRNAs, as does nuclease activity.
CC Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR
CC repeat sequences to help distinguish self versus nonself, as targets
CC within the bacterial CRISPR locus do not have PAMs. DNA strand
CC separation and heteroduplex formation starts at PAM sites; PAM
CC recognition is required for catalytic activity (PubMed:24476820).
CC Confers immunity against a plasmid with homology to the appropriate
CC CRISPR spacer sequences (CRISPR interference) (PubMed:21455174).
CC {ECO:0000269|PubMed:21455174, ECO:0000269|PubMed:22745249,
CC ECO:0000269|PubMed:24270795, ECO:0000269|PubMed:24476820,
CC ECO:0000269|PubMed:24505130, ECO:0000269|PubMed:24529477}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:22745249, ECO:0000269|PubMed:24505130};
CC Note=Endonuclease activity on target dsDNA requires Mg(2+)
CC (PubMed:22745249). The RuvC-like nuclease domain should have 2 divalent
CC cations, while the HNH domain should have 1. Crystals are often soaked
CC in MgCl(2) or MnCl(2+). {ECO:0000269|PubMed:22745249,
CC ECO:0000269|PubMed:24505130, ECO:0000269|PubMed:25079318};
CC -!- ACTIVITY REGULATION: Only has nuclease activity when bound to both
CC gRNAs (crRNA plus tracrRNA), which results in conformational changes in
CC the protein and formation of a central channel which binds target DNA
CC (PubMed:24505130). Also requires interaction with PAM to trigger
CC catalytic activity (PubMed:24476820). Nuclease activity is inhibited by
CC EDTA (PubMed:26841432). {ECO:0000269|PubMed:22745249,
CC ECO:0000269|PubMed:24476820, ECO:0000269|PubMed:24505130,
CC ECO:0000269|PubMed:26841432}.
CC -!- SUBUNIT: Monomer. Binds crRNA and tracrRNA. {ECO:0000255|HAMAP-
CC Rule:MF_01480, ECO:0000269|PubMed:24505130,
CC ECO:0000269|PubMed:24529477}.
CC -!- DOMAIN: Has 2 endonuclease domains. The discontinuous RuvC-like domain
CC (approximately residues 1-62, 718-765 and 925-1102) recognizes and
CC cleaves the target DNA noncomplementary to crRNA while the HNH nuclease
CC domain (residues 810-872) cleaves the target DNA complementary to crRNA
CC (PubMed:22745249, PubMed:24529477). {ECO:0000269|PubMed:22745249,
CC ECO:0000269|PubMed:24529477}.
CC -!- DOMAIN: Has a bilobed architecture with a recognition lobe (REC,
CC residues 60-718) and a discontinuous nuclease lobe (NUC, residues 1-59
CC and 719-1368) (PubMed:24529477, PubMed:24505130). The crRNA-target DNA
CC lies in a channel between the 2 lobes (PubMed:24529477,
CC PubMed:26841432). Binding of sgRNA induces large conformational changes
CC further enhanced by target DNA binding (PubMed:26113724,
CC PubMed:26841432). REC recognizes and binds differing regions of an
CC artificial sgRNA in a sequence-independent manner. Deletions of parts
CC of this lobe abolish nuclease activity (PubMed:24529477).
CC {ECO:0000269|PubMed:24505130, ECO:0000269|PubMed:24529477,
CC ECO:0000269|PubMed:26113724, ECO:0000269|PubMed:26841432}.
CC -!- DOMAIN: The PAM-interacting domain (PI domain, approximately residues
CC 1099-1368) recognizes the PAM motif; swapping the PI domain of this
CC enzyme with that from S.thermophilus St3Cas9 (AC Q03JI6) prevents
CC cleavage of DNA with the endogenous PAM site (5'-NGG-3') but confers
CC the ability to cleave DNA with the PAM site specific for St3 CRISPRs.
CC {ECO:0000269|PubMed:24529477}.
CC -!- DISRUPTION PHENOTYPE: Loss of correct processing of pre-crRNA and
CC tracrRNA. Loss of immunity against a plasmid with homology to CRISPR
CC spacer sequences. {ECO:0000269|PubMed:21455174}.
CC -!- BIOTECHNOLOGY: Coexpression of Cas9 with an artificial single guide RNA
CC (sgRNA) which fuses the crRNA with the tracrRNA in human cells has
CC shown it is possible to target and modify DNA sequences of interest
CC (PubMed:23287722, PubMed:23360966, PubMed:23386978). Cas9 plus the 2
CC sgRNAs have also been expressed individually in human and mouse cells
CC to achieve DNA targeting; cleavage efficiencies of the artificial sgRNA
CC were lower that those for systems with the 2 sgRNAs expressed
CC separately (PubMed:23287718). Microinjection of Cas9-encoding mRNA and
CC a synthetic sgRNA into zebrafish embryos induces targeted mutations
CC (PubMed:23360964). In all cases introduction of multiple sgRNAs leads
CC to multiplexed editing of genomic loci; DNA has also been inserted into
CC a mammalian locus of interest. In S.pneumoniae and E.coli it has been
CC used to generate markerless mutations; mutiple changes can be made
CC simultaneously (PubMed:23360965). Studies to make mutations that alter
CC the PAM-specificity and thus recognition possibilities have been made,
CC but are not annotated in this database (PubMed:26098369).
CC {ECO:0000269|PubMed:23287718, ECO:0000269|PubMed:23287722,
CC ECO:0000269|PubMed:23360964, ECO:0000269|PubMed:23360965,
CC ECO:0000269|PubMed:23360966, ECO:0000269|PubMed:23386978,
CC ECO:0000269|PubMed:26098369}.
CC -!- SIMILARITY: Belongs to the CRISPR-associated protein Cas9 family.
CC Subtype II-A subfamily. {ECO:0000305}.
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DR EMBL; AE004092; AAK33936.1; -; Genomic_DNA.
DR RefSeq; NP_269215.1; NC_002737.2.
DR PDB; 4CMP; X-ray; 2.62 A; A/B=1-1368.
DR PDB; 4CMQ; X-ray; 3.09 A; A/B=1-1368.
DR PDB; 4OO8; X-ray; 2.50 A; A/D=1-1368.
DR PDB; 4UN3; X-ray; 2.59 A; B=1-1368.
DR PDB; 4UN4; X-ray; 2.37 A; B=1-1368.
DR PDB; 4UN5; X-ray; 2.40 A; B=1-1368.
DR PDB; 4ZT0; X-ray; 2.90 A; A/C=1-1368.
DR PDB; 4ZT9; X-ray; 3.10 A; A/C=1-1368.
DR PDB; 5B2R; X-ray; 2.00 A; B=1-1368.
DR PDB; 5B2S; X-ray; 2.20 A; B=1-1368.
DR PDB; 5B2T; X-ray; 2.20 A; B=1-1368.
DR PDB; 5F9R; X-ray; 3.40 A; B=1-1368.
DR PDB; 5FQ5; X-ray; 2.14 A; B=1-1368.
DR PDB; 5FW1; X-ray; 2.50 A; B=1-1368.
DR PDB; 5FW2; X-ray; 2.68 A; B=1-1368.
DR PDB; 5FW3; X-ray; 2.70 A; B=1-1368.
DR PDB; 5VW1; X-ray; 2.60 A; A=1-1368.
DR PDB; 5VZL; EM; 3.90 A; A=1-1368.
DR PDB; 5XBL; X-ray; 3.05 A; A=1-1368.
DR PDB; 5Y36; EM; 5.20 A; A=1-1368.
DR PDB; 6AEB; X-ray; 3.00 A; B/F=1-1368.
DR PDB; 6AEG; X-ray; 2.70 A; B=1-1368.
DR PDB; 6AI6; X-ray; 2.70 A; A=1-1368.
DR PDB; 6IFO; X-ray; 3.31 A; A/B=1-1368.
DR PDB; 6K3Z; X-ray; 3.20 A; B=1-1368.
DR PDB; 6K4P; X-ray; 2.90 A; B=1-1368.
DR PDB; 6K4Q; X-ray; 2.70 A; B=1-1368.
DR PDB; 6K4S; X-ray; 3.01 A; B=1-1368.
DR PDB; 6K4U; X-ray; 3.20 A; B=1-1368.
DR PDB; 6K57; X-ray; 2.98 A; B=1-1368.
DR PDB; 6O0X; EM; 3.28 A; A=1-1368.
DR PDB; 6O0Y; EM; 3.37 A; A=1-1368.
DR PDB; 6O0Z; EM; 3.30 A; A=1-1368.
DR PDB; 6O56; X-ray; 1.90 A; A/B=775-908.
DR PDB; 6VPC; EM; 3.20 A; B=4-1364.
DR PDB; 7OX7; X-ray; 2.60 A; B=1-1368.
DR PDB; 7OX8; X-ray; 2.75 A; B=1-1368.
DR PDB; 7OX9; X-ray; 2.45 A; B=1-1368.
DR PDB; 7OXA; X-ray; 2.15 A; B=1-1368.
DR PDB; 7S4U; EM; 3.56 A; A=1-1368.
DR PDB; 7S4V; EM; 3.28 A; A=1-1368.
DR PDB; 7S4X; EM; 2.76 A; A=1-1368.
DR PDB; 7VK9; X-ray; 2.90 A; A=1-1368.
DR PDBsum; 4CMP; -.
DR PDBsum; 4CMQ; -.
DR PDBsum; 4OO8; -.
DR PDBsum; 4UN3; -.
DR PDBsum; 4UN4; -.
DR PDBsum; 4UN5; -.
DR PDBsum; 4ZT0; -.
DR PDBsum; 4ZT9; -.
DR PDBsum; 5B2R; -.
DR PDBsum; 5B2S; -.
DR PDBsum; 5B2T; -.
DR PDBsum; 5F9R; -.
DR PDBsum; 5FQ5; -.
DR PDBsum; 5FW1; -.
DR PDBsum; 5FW2; -.
DR PDBsum; 5FW3; -.
DR PDBsum; 5VW1; -.
DR PDBsum; 5VZL; -.
DR PDBsum; 5XBL; -.
DR PDBsum; 5Y36; -.
DR PDBsum; 6AEB; -.
DR PDBsum; 6AEG; -.
DR PDBsum; 6AI6; -.
DR PDBsum; 6IFO; -.
DR PDBsum; 6K3Z; -.
DR PDBsum; 6K4P; -.
DR PDBsum; 6K4Q; -.
DR PDBsum; 6K4S; -.
DR PDBsum; 6K4U; -.
DR PDBsum; 6K57; -.
DR PDBsum; 6O0X; -.
DR PDBsum; 6O0Y; -.
DR PDBsum; 6O0Z; -.
DR PDBsum; 6O56; -.
DR PDBsum; 6VPC; -.
DR PDBsum; 7OX7; -.
DR PDBsum; 7OX8; -.
DR PDBsum; 7OX9; -.
DR PDBsum; 7OXA; -.
DR PDBsum; 7S4U; -.
DR PDBsum; 7S4V; -.
DR PDBsum; 7S4X; -.
DR PDBsum; 7VK9; -.
DR AlphaFoldDB; Q99ZW2; -.
DR BMRB; Q99ZW2; -.
DR SMR; Q99ZW2; -.
DR DIP; DIP-61504N; -.
DR IntAct; Q99ZW2; 5.
DR STRING; 1314.HKU360_00834; -.
DR PaxDb; Q99ZW2; -.
DR PRIDE; Q99ZW2; -.
DR EnsemblBacteria; AAK33936; AAK33936; SPy_1046.
DR KEGG; spy:SPy_1046; -.
DR PATRIC; fig|160490.10.peg.902; -.
DR HOGENOM; CLU_005604_0_0_9; -.
DR OMA; TDRHSIK; -.
DR Proteomes; UP000000750; Chromosome.
DR GO; GO:0008408; F:3'-5' exonuclease activity; IDA:UniProtKB.
DR GO; GO:0003677; F:DNA binding; IDA:UniProtKB.
DR GO; GO:0004520; F:endodeoxyribonuclease activity; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-UniRule.
DR GO; GO:0043571; P:maintenance of CRISPR repeat elements; IDA:UniProtKB.
DR Gene3D; 3.30.420.10; -; 1.
DR HAMAP; MF_01480; Cas9; 1.
DR InterPro; IPR028629; Cas9.
DR InterPro; IPR032239; Cas9-BH.
DR InterPro; IPR032237; Cas9_PI.
DR InterPro; IPR032240; Cas9_REC.
DR InterPro; IPR033114; HNH_CAS9.
DR InterPro; IPR003615; HNH_nuc.
DR InterPro; IPR036397; RNaseH_sf.
DR Pfam; PF16593; Cas9-BH; 1.
DR Pfam; PF16595; Cas9_PI; 1.
DR Pfam; PF16592; Cas9_REC; 1.
DR Pfam; PF13395; HNH_4; 1.
DR TIGRFAMs; TIGR01865; cas_Csn1; 1.
DR PROSITE; PS51749; HNH_CAS9; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiviral defense; DNA-binding; Endonuclease; Exonuclease;
KW Hydrolase; Magnesium; Manganese; Metal-binding; Nuclease;
KW Reference proteome; RNA-binding.
FT CHAIN 1..1368
FT /note="CRISPR-associated endonuclease Cas9/Csn1"
FT /id="PRO_0000418437"
FT DOMAIN 770..921
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT REGION 1..62
FT /note="RuvC-I"
FT /evidence="ECO:0000303|PubMed:24505130"
FT REGION 56..718
FT /note="Recognition lobe"
FT /evidence="ECO:0000303|PubMed:24505130"
FT REGION 56..73
FT /note="ARM"
FT /evidence="ECO:0000303|PubMed:24505130"
FT REGION 718..765
FT /note="RuvC-II"
FT /evidence="ECO:0000303|PubMed:24505130"
FT REGION 925..1102
FT /note="RuvC-III"
FT /evidence="ECO:0000303|PubMed:24505130"
FT REGION 1099..1368
FT /note="PAM-interacting domain (PI)"
FT /evidence="ECO:0000303|PubMed:24529477"
FT MOTIF 1333..1335
FT /note="PAM substrate-binding"
FT /evidence="ECO:0000269|PubMed:25079318"
FT ACT_SITE 10
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000305|PubMed:24529477"
FT ACT_SITE 840
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000305|PubMed:24529477"
FT BINDING 10
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 10
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 762
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 766
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 766
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 983
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 1297
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 1328
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:24505130"
FT MUTAGEN 10
FT /note="D->A: Target DNA noncomplementary to the crRNA is
FT not cleaved; nickase activity. Processes guide RNAs. In
FT vivo, loss of Cas9-mediated CRISPR interference in plasmid
FT transformation. Able to bind guide RNAs and target DNA but
FT not cleave DNA; when associated with A-840."
FT /evidence="ECO:0000269|PubMed:22745249,
FT ECO:0000269|PubMed:24270795, ECO:0000269|PubMed:24529477"
FT MUTAGEN 15
FT /note="S->A: Decreased DNA cleavage."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 66
FT /note="R->A: Significantly decreased DNA cleavage."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 70
FT /note="R->A: No DNA cleavage."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 74
FT /note="R->A: Significantly decreased DNA cleavage."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 78
FT /note="R->A: Moderately decreased DNA cleavage."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 97..150
FT /note="Missing: No nuclease activity."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 165
FT /note="R->A: Moderately decreased DNA cleavage."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 175..307
FT /note="Missing: About 50% nuclease activity."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 312..409
FT /note="Missing: No nuclease activity."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 475..477
FT /note="PWN->AAA: Slight decrease in target DNA cleavage and
FT DNA-binding. Almost complete loss of DNA cleavage and
FT binding; when associated with 1125-A--A-1127."
FT /evidence="ECO:0000269|PubMed:24505130"
FT MUTAGEN 762
FT /note="E->A: Only cleaves 1 DNA strand, probably the
FT noncomplementary strand. Processes guide RNAs correctly. In
FT vivo, loss of Cas9-mediated CRISPR interference in plasmid
FT transformation."
FT /evidence="ECO:0000269|PubMed:24270795,
FT ECO:0000269|PubMed:24529477"
FT MUTAGEN 840
FT /note="H->A: Target DNA complementary to the crRNA is not
FT cleaved; nickase activity. In vivo, loss of Cas9-mediated
FT CRISPR interference in plasmid transformation. Able to
FT process and bind guide RNAs and target DNA but not cleave
FT DNA; when associated with A-10."
FT /evidence="ECO:0000269|PubMed:22745249,
FT ECO:0000269|PubMed:24270795, ECO:0000269|PubMed:24529477,
FT ECO:0000269|PubMed:25079318"
FT MUTAGEN 854
FT /note="N->A: Decreased DNA cleavage. Processes guide RNAs
FT correctly. In vivo, retains Cas9-mediated CRISPR
FT interference in plasmid transformation."
FT /evidence="ECO:0000269|PubMed:24270795,
FT ECO:0000269|PubMed:24529477"
FT MUTAGEN 863
FT /note="N->A: Only cleaves 1 DNA strand, probably the
FT complementary strand. Processes guide RNAs correctly. In
FT vivo, loss of Cas9-mediated CRISPR interference in plasmid
FT transformation."
FT /evidence="ECO:0000269|PubMed:24270795,
FT ECO:0000269|PubMed:24529477"
FT MUTAGEN 982..983
FT /note="HH->AA: Processes guide RNAs correctly."
FT /evidence="ECO:0000269|PubMed:24270795"
FT MUTAGEN 982
FT /note="H->A: Decreased DNA cleavage. In vivo, loss of Cas9-
FT mediated CRISPR interference in plasmid transformation."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 983
FT /note="H->A: Only cleaves 1 DNA strand, probably the
FT noncomplementary strand."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 986
FT /note="D->A: Only cleaves 1 DNA strand, probably the
FT noncomplementary strand. Processes guide RNAs correctly. In
FT vivo, loss of Cas9-mediated CRISPR interference in plasmid
FT transformation."
FT /evidence="ECO:0000269|PubMed:24270795,
FT ECO:0000269|PubMed:24529477"
FT MUTAGEN 1099..1368
FT /note="Missing: No nuclease activity."
FT /evidence="ECO:0000269|PubMed:24529477"
FT MUTAGEN 1125..1127
FT /note="DWD->AAA: No change in target DNA cleavage, slight
FT decrease in DNA-binding. Almost complete loss of DNA
FT cleavage and binding; when associated with 475-A--A-477."
FT /evidence="ECO:0000269|PubMed:24505130"
FT MUTAGEN 1132
FT /note="G->C: Probably inactivates protein."
FT /evidence="ECO:0000269|PubMed:23360965"
FT MUTAGEN 1333..1335
FT /note="RKR->AKA: Nearly complete loss of target DNA
FT cleavage."
FT /evidence="ECO:0000269|PubMed:25079318"
FT MUTAGEN 1333
FT /note="R->A: Dramatically reduced target DNA binding,
FT slightly decreased target cleavage."
FT /evidence="ECO:0000269|PubMed:25079318"
FT MUTAGEN 1335
FT /note="R->A: Dramatically reduced target DNA binding,
FT slightly decreased target cleavage."
FT /evidence="ECO:0000269|PubMed:25079318"
FT STRAND 6..11
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 13..21
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 23..25
FT /evidence="ECO:0007829|PDB:6K4U"
FT STRAND 29..39
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 42..52
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 60..93
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 97..102
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 103..105
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 108..110
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 117..119
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 122..131
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 135..144
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 151..163
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 178..180
FT /evidence="ECO:0007829|PDB:4UN3"
FT HELIX 185..195
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 196..198
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 205..207
FT /evidence="ECO:0007829|PDB:6K4S"
FT HELIX 208..212
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 214..216
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 218..227
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 229..231
FT /evidence="ECO:0007829|PDB:7OX9"
FT STRAND 234..236
FT /evidence="ECO:0007829|PDB:6K57"
FT HELIX 237..246
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 253..257
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 266..268
FT /evidence="ECO:0007829|PDB:5FQ5"
FT HELIX 271..282
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 284..286
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 287..299
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 300..305
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 310..312
FT /evidence="ECO:0007829|PDB:7OXA"
FT HELIX 316..342
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 345..351
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 356..358
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 359..363
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 369..382
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 383..385
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 387..394
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 402..404
FT /evidence="ECO:0007829|PDB:5VW1"
FT HELIX 405..409
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 412..426
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 427..429
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 431..435
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 437..445
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 450..452
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 467..471
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 475..477
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 478..481
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 484..493
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 500..502
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 505..509
FT /evidence="ECO:0007829|PDB:7OX8"
FT HELIX 513..524
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 528..530
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 532..534
FT /evidence="ECO:0007829|PDB:4ZT0"
FT HELIX 542..551
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 552..555
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 556..558
FT /evidence="ECO:0007829|PDB:5FW1"
FT HELIX 561..567
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 568..573
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 579..582
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 584..586
FT /evidence="ECO:0007829|PDB:5FQ5"
FT HELIX 592..601
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 604..608
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 610..612
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 613..625
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 629..636
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 637..642
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 645..652
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 659..663
FT /evidence="ECO:0007829|PDB:7OX8"
FT HELIX 664..668
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 673..675
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 679..684
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 687..689
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 693..698
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 700..703
FT /evidence="ECO:0007829|PDB:5FW1"
FT HELIX 704..711
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 720..725
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 727..729
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 731..750
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 751..753
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 757..763
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 771..774
FT /evidence="ECO:0007829|PDB:4ZT0"
FT HELIX 777..791
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 795..798
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 803..807
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 809..816
FT /evidence="ECO:0007829|PDB:6O56"
FT TURN 817..819
FT /evidence="ECO:0007829|PDB:6O56"
FT STRAND 822..827
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 830..835
FT /evidence="ECO:0007829|PDB:6O56"
FT STRAND 836..842
FT /evidence="ECO:0007829|PDB:6O56"
FT TURN 844..846
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 852..854
FT /evidence="ECO:0007829|PDB:6O56"
FT STRAND 855..859
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 861..864
FT /evidence="ECO:0007829|PDB:6O56"
FT STRAND 867..871
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 873..888
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 894..900
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 902..905
FT /evidence="ECO:0007829|PDB:6O56"
FT HELIX 910..921
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 926..939
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 945..947
FT /evidence="ECO:0007829|PDB:4ZT0"
FT STRAND 953..957
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 960..969
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 976..978
FT /evidence="ECO:0007829|PDB:5FQ5"
FT HELIX 981..1000
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1002..1004
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1005..1008
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1009..1011
FT /evidence="ECO:0007829|PDB:5VW1"
FT HELIX 1018..1021
FT /evidence="ECO:0007829|PDB:5VW1"
FT HELIX 1026..1028
FT /evidence="ECO:0007829|PDB:5VW1"
FT HELIX 1032..1040
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1042..1046
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1048..1051
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1053..1055
FT /evidence="ECO:0007829|PDB:6AEG"
FT STRAND 1057..1059
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1062..1065
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1067..1069
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1072..1075
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1076..1078
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1079..1087
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1093..1096
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1105..1111
FT /evidence="ECO:0007829|PDB:6AEG"
FT STRAND 1115..1117
FT /evidence="ECO:0007829|PDB:4OO8"
FT STRAND 1120..1123
FT /evidence="ECO:0007829|PDB:4UN4"
FT HELIX 1128..1131
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1133..1137
FT /evidence="ECO:0007829|PDB:6AEG"
FT STRAND 1139..1151
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1152..1155
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1156..1167
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1168..1170
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1171..1176
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1178..1185
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1187..1189
FT /evidence="ECO:0007829|PDB:5FQ5"
FT HELIX 1192..1194
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1196..1198
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1203..1205
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1207..1209
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1211..1218
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1220..1222
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1230..1240
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1242..1244
FT /evidence="ECO:0007829|PDB:5VW1"
FT HELIX 1251..1261
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1262..1264
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1265..1280
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1284..1296
FT /evidence="ECO:0007829|PDB:5B2R"
FT TURN 1297..1299
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1302..1316
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1317..1320
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1324..1326
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1329..1331
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1334..1336
FT /evidence="ECO:0007829|PDB:6IFO"
FT HELIX 1340..1344
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1345..1350
FT /evidence="ECO:0007829|PDB:5B2R"
FT STRAND 1352..1354
FT /evidence="ECO:0007829|PDB:4UN4"
FT STRAND 1356..1361
FT /evidence="ECO:0007829|PDB:5B2R"
FT HELIX 1362..1364
FT /evidence="ECO:0007829|PDB:5B2R"
SQ SEQUENCE 1368 AA; 158441 MW; 07D04F0B5965762F CRC64;
MDKKYSIGLD IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE
ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG
NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD
VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN
LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI
LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA
GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH
AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE
VVDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL
SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI
IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG
RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL
HEHIANLAGS PAIKKGILQT VKVVDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER
MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDH
IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEVVKKMK NYWRQLLNAK LITQRKFDNL
TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS
KLVSDFRKDF QFYKVREINN YHHAHDAYLN AVVGTALIKK YPKLESEFVY GDYKVYDVRK
MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF
ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA
YSVLVVAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK
YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE
QHKHYLDEII EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA
PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD
