F SK TGI A QF C L DYPAEKV NP DQ T AK GT F PY D L DPF WKTIK HESRKH LEG F H

VI L VV V S N E I N EL I V V VIA HDDN.PY G N IV GK N E Y L E I NFNGIRIKTD HSL DKK FE QN TS E A K CE EK D L D II I VI L S Q E V T DL L I M VVV EHPE.TP G N IT ST K E R L T Q F........D QKK DNR VA QA SV G Q E VD IH Q L N IL A LV I T N D I N EM L V V IVV EKANDVH S H YT GK N K D F I G D....RMKTN HDK AAI DS KD KK N E E AK IE N F D

Figure S4

1 10 20 30 40 50 60 70 80 90 100

FnCpf1 M F N Y SKTLRF IP GKT I D R YK K D L L K N I S SL L I D A II K FI V I Q Y D T K IYQE V K E Q LEN KARGL L EK AKD K Q YHQF EEI SS C SEDL NYSDV F LKKS DD LQKDFKSAKD I KQ AsCpf1 M F N Y SKTLRF IP GKT I D R YK K D L L K N I T QV L I E L II R YA V L S Y E T R QFEG T L E Q LKH QEQGF E KA NDH E P IYKT DQC QL Q DWEN AAIDS R EKTE TR ...ALIEEQA Y NA LbCpf1 M F N Y SKTLRF IP GKT I D R YK K D L L K N I S SL A L E V LL R FI I L N F E N R KLEK T C K V QEN DNKRL V EK AED G K YYLS NDV HS K KN.. NYISL R KTRT KE ..KELENLEI L KE

110 120 130 140 150 160 170 180 190 200

FnCpf1 E K LF SF TT F GF NR N S T I R F L II W E VY V SEYIKD.............S K N NQN IDAKKGQESDLILWLKQSKDNGIELFKANSDITDIDEALE K KG Y K H K SNDIP S IY I DAsCpf1 E K LF SF TT F GF NR N S T I R Y L LL F E VF V HDYFIGRTDNLTDAINKRHA I G KAE FNG....................KVLKQLGTVTTTEHENA R DK Y S Y K AEDIS A PH I QLbCpf1 E K LF SF TT F GF NR N S T I R Y I LV F D MF I AKAFKG.............N G S KKD IET........................ILPEFLDDKDEIA N NG A T F E EEAKS S AF C N

........................

210 220 230 240 250 260 270 280 290 300 310

FnCpf1 N N F N L Q I N GG K KG NE NL Q D KF Y L LDE F N F II I I LP LE KAK ES KDKAPEAINYEQIKKDLAEELTFDIDYKTSEVNQRVFS V EIAN NY SG TK T KFVNGENT R Y YS QINDKAsCpf1 N N F N L Q I N GG K KG NE NL Q D KF F L IEE Y T Y LL L L FP KE CHI TR ITAVP...SLREHFENVKKAIGIFVSTS.......... V SFPF QL TQ DL Q ISREAGTE I V AI KNDETLbCpf1 N N F N L Q I N GG K KG NE NL Q E RY F V VED F T Y II L I LT IS MDI EK D......AIFDKHEVQEIKEKILNSDYD.......... F EGEF FV EG DV A .FVTESGE I Y YN KTKQK

......

.... 320 330 340 350 360 370 380 390 400 410

FnCpf1 L KQ LSD SF D V LF S K F I S D D V S L N L L V Y I T....LKKY MSV TE K VI KLED S TTMQSFYEQIAAFKTVEEK IKET SL DDLKAQKLDLSKIYFK DKS TD QQ FDD SV GTAVLEAsCpf1 L KQ LSD SF D V LF S R F I T E E I N A S L I L W L AHIIASLPH FIP RN L IL EFKS E QSFCKYKT......LLRNE VLET EA NELNSIDLT...HIFI HKK ET SA CDH DT R.....LbCpf1 L KQ LSD SF D V LF S K Y V S E E L S L N I I I W I .....LPKF P.. RE L YG GYTS E EVFRNTLN.....KNSEIF SIKK EK KNFDEYSS..AGIFVK GPA ST KD FGE NV R.....

. 420 430 440 450 460 470 480 490 500 510 520

FnCpf1 I K I I DI A D AI YITQQIAPKNLDNPSKKEQEL AKKTEKAKYLSLET K.LALEEFNKHR DKQCRFEEIL NFAAIPM FDEIAQNK NLAQISIKYQNQGKKDLLQASAEDDVK AsCpf1 I K I V EI A D IL ..............NALYERR SELTGKITKSAKEK QRSLKHEDINLQ ISAAGKELSE FKQKTSE LSHAHAAL QPLPTTLKKQ.............EEKE LbCpf1 I K V I DA I E IM ....DKWNAEYDDIHLKKKAV TEKYEDDRRKSFKK GSFSLEQLQEYA DLSVVEKLKE IIQKVDE YKVYGSS. KLFDADFVLEKSLKK......NDAVVA

530 540 550 560 570 580 590 600 610 620 630

FnCpf1 LD F D F Y RNY T KPYS KFKL F GWD KE A YYL M K Q L L I I E T A L V DL TNNLLHK KI HISQSEDKANILDK EH YLVFEECYFE ANIVPL NK Q D N ENS L N KN PDNT I FIKDDK G N .AsCpf1 LD F D F Y RNY T KPYS KFKL F GWD KE A YYL M K S L M A A E T A L I SQ LLGLYHL DW AVDESN......EV PE SARLTGIKLE EPSLSF NK K V N QMP L S VN KNNG I FVKNGL G P QLbCpf1 LD F D F Y RNY T KPYS KFKL F GWD KE A YYL M K S I L I V D Q M I I DL VKSFENY KA FGEGKE.....TNR ES YGDFVLAYDI LKVDHI DA Q K Y QNP F G KD TDYR T LRYGSK A D .

.

640 650 660 670 680 690 700 710 720

FnCpf1 K K Y P KM PK K K IDF E Y I L S I S I NNKIFDDKAIK NKGEG K V KLL GAN VFF A S KFYNP EDILR RNHSTHTKNGSPQKGYEK....................FEFNIEDCR F YAsCpf1 K K Y P KM PK K K IDF E F M I Q V T L GRYKALSFEPT KTSEG D Y DYF DAA CST L A TAHFQ HTTPI LSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALC W TLbCpf1 K K Y P KM PK K K IDF D Y I L S M S I YAKCLQKIDKD VNGN. E N KLL GPN VFF K W AYYNP EDIQK YKNGTFKKGD.............................MFNLNDCH L F

730 740 750 760 770 780 790 800 810 820 830

FnCpf1 S Y Y E Y F V GKLY FQIYNKDF G PNLHT Y LF N L G AELF K I K T I F V KLT I I V L K L W N QS HPEWK.DFGFRFSD QR NS DE R ENQG EN SESY DS NQ SAYS R KA DER LQDVVYK E YAsCpf1 S Y Y E Y F V GKLY FQIYNKDF G PNLHT Y LF N L G AELF R L K S L Y L HIS I I A L H L W N DF YTKTTSIDLSSLRP SQ KD GE A NPLL QR AEKE MD ET AKGH K TG SPE LAKTSIK Q YLbCpf1 S Y Y E Y F V GKLY FQIYNKDF G PNLHT Y LF N L G AELF K I R T I F V KVS A V L M H M F S DS YPKWSNAYDFNFSE EK KD AG R EEQG ES SKKE DK EE SDKS T KL DEN HGQIR.. G M

840 850 860 870 880 890 900

FnCpf1 R S K H NK K KD RF D H PI N K N L I A IA DL K T F I I L KQ IP K.. T P KEA NKDNP K..................................ESVFEY I E K FF C T FKSSG.AN F DE N L AsCpf1 R S K H NK K KD RF D H PI N K N L M L ML EI R T F L V A PK RM R.. A R GEK KLKDQ TPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSH I S K FF V T YQAANSPS F QR N Y LbCpf1 R S K H NK K KD RF D H PI N K N L V A IA DV K S Y I V V RA LK EEL V P NSP NPDNP K..................................TTTLSY Y E Q EL I A KCPKN.IF I TE R L

910 920 930 940 950 960 970 980 990 1000

FnCpf1 K IDRGER L Y D G I Q N I Y L EK R AR W IK K GY SQV H I L Y A E LNAsCpf1 K IDRGER L Y D G I Q N I Y L EK R AR W IK K GY SQV H I L Y A E LNLbCpf1 K IDRGER L Y D G I Q N I Y L EK R AR W IK K GY SQV H I L Y A E LN

1010 1020 1030 1040 1050 1060 1070 1080 1090 1100 1110

FnCpf1 V E L V L T T IY V L S I F RG .FK Q KL Y F DNEFDKT RA AP E KK GK I GF C V N.Q YPKYE VSKSQE FSK K CAsCpf1 A D L V L S S LF V V N L LbCpf1 V D M A I S N IF I L S I S NS .VK Q KF Y D KSNPCAT KG NK E KS ST F WL D S N.L KTKYT IADSKK ISS R M

1120 1130 1140 1150 1160 1170 1180 1190 1200

FnCpf1 Y F F K G R L I G I D D L K EV Y I NLDKGY EFSFDY..................KN G.. KAAKG WTIASF S INFRNSD NHNWDTR YPTKELEK LKDYS E GH EC KAA CGES KKFFAAsCpf1 Y F F K G R L I G I D E I R DL F L DVKTGD ILHFKMNRNLSFQRGLPGFMPAWDIV EKN TQFDA GTPFIA K VPVIENH .FTGRYR YPANELIA LEEKG V RD SN LPK LEND SHAIDLbCpf1 Y F F K G R L I G I D D I K EV Y L VPEEDL EFALDY..................KN SRT ADYIK WKLYSY N RIFRNPK NNVFDWE CLTSAYKE FNKYG N QQ D. RAL CEQS KAFYS

α1

1210 1220 1230 1240 1250 1260 1270 1280 1290 1300

FnCpf1 VL I YL F LL E L EY FV KLTS NT KTG. EL I ADVN NF ....QAPKNM QD H GL GLM GRI NNQ G ..K NLV K E FE NRNN. AsCpf1 LI V YI F LL D L DW YI TMVA RS NAA. GE N RDLN VC ....FQNPEW MD H AL GQL NHL ESK L ... QNG S Q LA ELRN. LbCpf1 LM M FL Y AI D V EW YA SFMA SL ITGR DV I KNSD IF NYEAQENAIL KN N AR VLW GQF KAE E LDK KIA S K LE TSVKH

RuvC I RuvC II

RuvC II

Zn Finger-like

RuvC III

Bridge Helix

GFK R EK VYQ EKMLI KLN V K GG L YQ T F F M Q G Y PA TSKI P TGFV F FD GFK R EK VYQ EKMLI KLN V K GG L YQ T F F M Q G Y PA TSKI P TGFV F FD GFK R EK VYQ EKMLI KLN V K GG L YQ T F F M Q G Y PA TSKI P TGFV F FD

LQMRNS T D SPV G DSR P ADANGAY I K K K I N Q LQMRNS T D SPV G DSR P ADANGAY I K K K I N Q LQMRNS T D SPV G DSR P ADANGAY I K K K I N Q