In vitro validation methods for car-t cell therapies

Abstract

The invention includes improved methods and compositions for detecting balanced reciprocal chromosomal translocations in the genome of an engineered cell. Methods are useful as quality control for CAR-T cell or CAR-NK cell preparations.

Description

[0001]. - - e onus a.caon camsroro..rovsona acaon sera no. , one. - n . ormea a ancaon conans aeuencesn wcaseen sume eecronca onu, ,s nsameree “ncororae reerencens enre.a . co, creae conanens . esar o.e.x sme”c ancason, a esn sze.e seuencesn erenns enre. nserencororae reerence reaeo a meenoven oo vna reaaneoeneecae e on cneeereermam.ore secca,envenon eecn anuann cromosomaransocaonsne ceunse. ces usen ceeraes reeae seromse o aove ceerasoever enneeremmune cesa w ennaeee anesroumor ces or oerarmu orsease cesneaen’so.mmune ce cs ao “oxvcoenaersssrouee oe ce ansroen, man aove ces are armee w saen- co m-e meer-ac crne an a.”oenv reecemmorusne cess conmserrese mnuoee ceeunoamree enoms.rs,e e cesncue one or moreenomc mocaonsa enanceoenc o e.econ, e ces.uc aona mocaons mancueenenocous w our wroon survva o enenoc-ns.oue as ssem aowsorrecseenome enneoeurn a vona secc cus,aren mue ses wneenomeeaves cesroneoenao vmerc ^ . r eenarrnaenerenmen ans- suc as,or- ex caemse w, crowmoosomaransocaons.aseen reorea socoma comex ass aonaenenocousa sencee maor arorn cromosomar an ass resusn. o enneere ces nuceasesor smuaneousannsococ-ano anns.asaes eer en a.r,evensransoomcanonnsn meren - ces,enomeoo, uex-ee rearaonsoer eensusre a neeor r:er.ouce meos o vaan enneeremmune ce aens. sae anoenc oerearaonsroroemen amnsereo recrocarnan osnoec eamonomen,e.n:ven.onns a a meenoome o o aene ecneresence o aaance meo comrsn: conacn a same nneere mammaan ce,e amcaonrmers comrsn one or more enneere ces w seuencen , wereners amcaonrmers caae o rzno a seuencenee enee,n aen, an aoem seecroasne, a amnercoarmonnr amne arms cacaaoen oerozno a can reacon orouce an amcon, werenere merasencave oeresence or a sence or asence oe amcons asence oe amconseseenccee oeeraencsnocaouno.rensc seonmcee o em aoameens,eresence or emomens,eransocaonseween a se eroe.n some emomens,ers amcaonrmerne ene ane ene.n some amcaonrmer consss e consss essena o :,e secon :.n some emossmena o :, aneroe consss essena o :,e seco ens,ers amcaonrmer consss essena o consss n amcaonrmer consss essena o :, aneroe amn ess aen coanro o nucec ac se:ue.ncne soremseen emnoemeennosm,e oe m weoeur cer comrses enome o enneere ces aneecneresence o es anne roe.n some emomens,e e conro nucec ac w a conro roeeecneransocaon meou.r:er. com anrsese coumoreasrcennce oeueorersoceenceee ocne ^ . ameo cnon ernon neuecreec c aecs.,ne soremeermnne numer oransocaons .:. n one emomee emomens,e sa . aance recrocaransocano,nenvenon.s: a re.aconn a mexnoumreeo or aene ecnnneereer mesaemnmcea oan a cee,rse a remaconca moxnurrem coerm carsane: o nucecr acznssoo aae seureonm one or more enneere ces, secon amcaonrmer caae orzno a seuencceene ene, ane emomens,ers amcaonrmer consss essen ne ene.n some amcaonrmer consss essena o :a. on some e: ane secon amcaonrmer consss essena o : ane seconm aomens,ers consss essena o :.n some emomens,e reacmocaonrmer comrses one or moreaeeroe seecerom a se n mxureurer : an a seuence conssn essena o uence conssn essena o mxureurer comrses ars an :.n some emomens,e reacon a conro nucesc, a anc a s secon conro amcaonrmers caae o amn enneere ce ceonueronceroreese cnaaneeeeecnonme am o wcaone o cese a cnonrno neuceenco amce o. rec n one emomen,envenons aoreecneresence o aaance cormocarsrnan:socearosn amcao.n:rm.en aenome o an enneere mammaan ce,e en r caae orzno a seuencene n seo,m aen emeo secmoenns a,mercsa aomnrmcaero canarmee or consrssz esnseno aa se ouencene: anene. secon amcaonrmer consss essena o :.n some emomens e amcaonrmer consss essena o : ane secon amcao,nerrs consss essena o :.n some emomens,eurer comrse meraeeroes seecerom a seuence conssn esse s one or more conssn essena o :.n some emonmaens o,e : an a seuence an secon conro amcaonrmers caae o a urer comrses ars resenneenome o w e ces an mn a conro nucec ac seuence caaeeecn amcao neenome o enneere ces, an a conroroe comrses nuceoserosn oaes,e a conerromo nsucaeec a ncuc.ecn s aocme eommoermaseen asn,euerusr aner ^ . cuoraceror cso nmecressseasr reaoe sunssaonr enrzemaranc s aecuvenc oneraorems aenrase se.nue snocmne em nucoecm aecnss,.e ..eesc aaram oeaance recroca cromosomaransocaon useraesee meeson o oee anssvaenoorn.eecn ... een e eoo ermonws “anoevnons an unersann osscosure. c e ce” reerso a cea caneeneca moeor usen a ceeras.reamen.xames o aove cesncue-ces, macroaes, an nauraer eneca moeerem “ ccees.ereaer”m re “aerosovee cereaemraen o as”e raeseer osros aoreerra uzes eneca moe aove ces.xames o ncue- aa uses eraes, nauraer ceeraes an - ceeraes, - ce eerm “moce” reerso aeuoce ceaesraaes. ssem.mocesncue-ce+ +r oe vereraemmune ammaea-ces, an reuaor-sce susc. amsoces or asoncu-ece nasu,ra aaerea-c cees, nauraer ces, conenuce er ces, an anen s, s, suc asenrc ces.moces asonc resenn ces eerm “nucec ac” or “ ueumornranmocess. or ronuc onuceoe” reersoeoxronucec acs seccaecm aces,eerm an encoommearsssese nrueon eer sne- oroue-sraneorm.ness nuceoesaave smarnnroerecse asc aec rese croenncaen nuncenown anaoues o nauran a manner smaro naura occurrn nuceoes.ness oerwcs aecn anca aere, m aeaarocuze nucec ac seuence asomc encomasses co ar eenerae co nservave moe varansereoe.., ase seuenceo enx sucsunonsca, aee.es, orooues,s, an comemenar seuences as we ^ . ase oeraeneremo “umro conveen nuocnaeoe”sa ussees coenrsesnno oesc areneo as nnuec,eouanoesn w a an cosne.eoxurac aneoxnosne are moe nuceo ne,mne nuceoes ma comrse mocaons oe suar moe.n , mesone.oe comrse a suar moe oeraeoxrose.or exame, r nuceoes “moe nuceoes”n .oe nuceoes ma onuceoes are consere me-, a-e-, a-me-, a-e-, c aom,r-se moease moees.- , a-ron-, a-ron-, a-ron-, aam-rnooaurrne, a-ron- roaramno-, a-roaramno-, a-roaramno-,amno-, a- eoxxanosne, arazoormne anao, aseuo-, a n a-eaza-- me o-,′--me o-, an-e-, an-merorroe, a nronoe,′-- ee are useu moe nuceoesn amcaonrmers-, enzurnes an ac mocaon anormaon ormerme a reuce non-secc nucec eerm “rmer” reerso an oorns,u sceeeo.e. wa.co.,,. sne-sraneemae nucec ac moecue an nso a secc reon o a omerase-meae enzmac reacon naes nucec ac sness va a nuceoes anreera com . ca, armer comrseseweran aou secca rzeso rseseweran aou nuceoes.are-seccrmer rzaon conons ca aareonuceoe uner rzaon conons.uc amcaonuer mnrnsc-ue,,u m are nomeo,rzaonnsoermaen oe,rmer., aerec ° com a aememenaerraurse n oo a4reou24ure °, oro m aeoru, mer ° m o s.eecencan4, on.e eare seuence unerrzaon conons.eerm “susana co nooeeree o comemenar suceno aceve secc memenar reers conons.n aonoeare-nn reon, arme rzaon unerrzaon reons,cane’-oron.e aona re r maave one or more aona se or aarcoe.eresence ons mancue a unversarmernn e a oermeror oze aona reonsne’-oronoes no neave aec eerm “amcaone coonsonneneare unerrzaon conons. reacone.., amca s” reerso cononsn a nucec ac amcaon oermers.eerms “amoncon”a an ao “wamorcaronzaroonuc an” reeemroa ae n-ueceenc aecn e mxoenescuone ^ . oan c vonraon asm a orc aaroanmen oeare nucec ac seuence anasorme aserouc eerm “ an suae amcaon meo. nn se.nversaunvremrsearrnmner” s reeesr csano ae nramuerra oar a crancar sezueeno a unversarmero aare seuen arcoes caneenrc eme ra “n cesca ae e aar nco eon-e o”ar r r mee oe- rrs ese aoc n a nc uuc m oean acn ae ocr. u se nuuecnceeoaes caonne.eece anene.oave aeas a mnmum numer oerencesro arcoes areesne cane unueo eac moecu m oerarcoesn aouaon.arcoes moecuesne same en a same or unueoe same ane sare mue arcoeaenes a. sameeer omr a “m souurcee oxeen samer,”e. “s su”c or, a “sa omr suesaarcnoae” r aeero a arcoeonuceoesrom a sne source or same w sare an o , we a, or susana ae.., aeas or , arcoeoe same seuence eren sources or sames wave aeren arcoe nuceoesrom eren sourcesavneren s cane seuence.onuceoesrom e samenormaon enco mxe an seuencenarae we manann “,” reero aa ene arcoe.eerm “unue moecuarener” or susana aercoeaenes aonuceoeo wcs aace.ca, a, or onuceoes are. u.n, aeas or , arcoesn a mxure o arcoe ces resuneronmve mnuonuen. evoenvoesmee eecsn a s ceo-msmecon cc nruomosomaransocaonn enneeres a meo o assessn weer ar ceases.orerecse,envenon nserenoe ce receeoarra aona o - ces or - cea comrseenacvaon oe enesreeora consan ocus anurer comrsnransocaoneween s a ceran amoun o aaance cromosoma oe ocus anee ocuosc ane ocus resunrom smuaneous en se oue asu ss.sem aowsorrecseenome enneern va ver rearcrance cmuesn,s saurce ans,o mru examee s, cesro wmosonmaeranesnoocmeeaves cesroneoenomc ecenecn ona aenveernsome o en. reaeraw reaasrna a moan wms am.a maons cesnv reosvuesnn reessara on ^ . o nesnu eorme oe s n eran onrea e exsoenous s as ecucc uuerna a ces e aoem oone eos eon sursro e reu sccoe emnnaeoe cneuar.-as eno re nsuucesasnesruercnse accuraeawas o non-omooous enonn an me.owever,essonn are asonvoveseeue anreene mcro-omoo-meae en -as meaeenome en,rensene.,:..ur rearawa cocesn awas resun cromosomaransoc ermore,e error-rone o a ce.aseen reor aonn case o mue cusnroucenoeenome nocousa sencee meaoa enneern an- - ces wwo aonaene resusn. o enneere cerssoarcoomra comex ass an ass omnn eren nucea n cromosomaransocaons.aser e a.,ransocaonsn sesor smuaneousnoc-n anase enrevens ecau mseu oenxn-aeee unre-ca ces, oenome roeo,:. sueceo mueenome es, enneeremmun arnvn cesaaveeen - cerearaons reureua co e cerearaonssuc as - ces or ne wao ensureaee nroroceureseoree cane usene cnc. seuencn.ouease,nomse as oroa encnseemree c coenssum arenn aanc resuor ceoneu scacr wcoee o-enome numer o enneere ces.o aresssroem,envenorsa aare a sma auo oe cerearaono ra eec cromoveevse a meo o usnwo cu sesne ceuarenome.n s somaransocaonseweene eresence oeaance ome emomens,envenons a meo oeecn oc cromosomaransocaoneweene ce receor aa consan ae as aus va ane ocusneenome o a mammaan ce.e meo cane - ces orao-n or c aesuaror coon arom mnesaseurrneore a ceresaora aoan oen e.nene mereeo ce casne a.., e aeo enneere cese.., - ces or - cessoaerom a receso aen asar o monorneaen’s resonseoera we enneere - cn - ces.e meos acaeo au es or mmun ooous - or - ceouaons ase ceserverom aaen an uon en, renusenoeaen.e meosmmou ane cceaseerov aeoreonmecea-o onrors an- ma cee reaouoarnonussoon-noe- asneareenar.aons o ^ . aance crnomroosaomaermrasn,socea moenoew ceoemnrseseoc suesso on aremec on cromeosresence oe ocuson arm o cromosome.ore secca,e meo comome ane rmers anroes seccoe seuencesne ene ane resese use o e cu ses.n some emomens,e meourer comrse ne aacenoransocaonsne cerearaon.n some emo suanne numer o or more reerenceenoe mens,euannsn reerenceo one envenon cnoomusr sseesuenece ussene ceuarenome. a cane useunmm ouman or mammaan ces suc asmmune ces - ces, n unoera.emmune ces oensannvenonncue ces, enneere auraer ces,nuce nauraer ces, macroaes aner ner sovmaev eems soucm aesns,-e c ces, - ces anee. cuure,.e., mana es anaze e meo oenvenon areresenn n somen eemno amreonws, mee ceums o aurse oeumano. comrses a suae excen eresenn aeraeuc comosona aso anmcroa a comrsn one or more o caroraes,noranc sas, cern, veeaeen os,s a,nooxsaons, sus,r saucraancsa,nsu, suersa,rs a,c as,ases, waer, acoos,oos, n some emomens,e ces are rerevenromervaze suars. receve annuson o enneeremmune ces aaen woasrevous emomens,e ces arerese asar oereamen remen.n some ereraoo an coroon amonmocesa aresoaeromoo,ncun m noes, an m , orrommo orans suc asemus,one marrow, mocesrom suucosa-assocaemossues .ecnuesorsoanecnues a cssues are wenownne ar, see, e..,m,.. eress nn s coemeua ermmomumnoemnos,uaon,er.er.:-. secc,reuenc, an esoaemoces are caracerzenerms o enrceor se uncon.n some emomens,esoaemoceouaons n socmec e smusoesm oen ces,s, seucso as ces or ces. suses o ces, suc as+ +ae+moc+eouaons enrceor secc –ncocs : .,n som,e emo, omre .ee, e..,an e a.,o.era enrceor+enoe reresenn cesn.s,esoaemoceouaons ^ . aen’s son somsse emomense - ces or - ces maesoaerom a o us oeruea,n or aoo soumor. soumor same mae oane os.e ce-ree erverom ma aso comrsee - ces, - ces or nucec acs swea suc ces, e.., urne, suum,oo serum,m, sava, suum, eura,eaur, c,e cresrcosuna,ue,,a asmrncouc,un, snovau,ercarau,eroneau, n some esnau, aneca sames. rom aaen wo emaomens,e meo oenvenons aeo ce-ree oane ma comrse ce-rseeeenreae w enneere - an - ces an woseoonormave oneenec caraceerverome - or - ces ansereore exracerome anv saemnoen.nveovoesr s ms oanc o nuuca suc ecn c acseos ea. xerac nuocnec acsaaveeensoae or amroo e a.,oecuaronn:aoraoranu are wenownne ar.ee. aoraorress:ewor,... vare os area c,om,n.,ornaror nucec acs or romooca samese..,merca avaaeor exracn euencnouons,easanon,a., oscencesaxressxracoce ecnooesason,sc.enrasems,nneaos,nno. o,a.,cenre monusn,ex.oaaoraoresercues,a., an aenaenca,a., eecen seom mee emooesmcrenes,eere snecnvocveesne cecav maoe oca ono m eno o ore. r m ao creeue.arenomeoe secc sesne ceuarenome w a seuence secc eno .,wo or more exoenous nucec ac“onor nucec nucease.on ceavae, an emomens,ensere n ac” canenroucenoe ceave se.n some emomens, “nae” nuceucce acc asc co emncrosens a ex comeneroucs anroeenn re ecxereosrson con.srnuc som aenroucenoe ce eecrooraon asescre e..,n..aeno.,, re emomens, a vecors use, e.., a vra vecorervero .n some conssn o an aenovruse an an aeno m a vrus seeceromerou assocae vrus , a s vruse, a rerovrus, aenvrus, an aeno- , aner man vrus -, vaccna vrus,ena vrus,sen-arr vrus vrusnces smex vrus .n some emomens,e vruss an vrus aneranscron anororraanessaoen o neucec ac ex sreessueonnce conavsrnuc aanee eemensom neoceossar armosr ^ . emomooomoeunss,o eace soemoueonce a orm coemersseresnse-rona sseeane ceuarenome.n some asears o seuence caae orznoe rs or- asears or aounseron oe enesesre. seuencene ceuarenome were encoene some emoocums.ense,e enonuucseearseese au naurcence arsca-ceueor enaonnucroease eeasenomcocussoun manroarocenomes anrove mc ooren nucec acs.rucure, nom s ressanceonvasonnaarova e a., encaure an casscaon o oc are revewe crooo vouon an casscaon oe -as ssems.aureevewsnersace wr.e, s aune acers.ca:-. e ocoucsusn acsuoensc au neusm coern o s seourenc reeeas reuar assocaeasenes. sacer-reea seuence un enc sor - a maure crs arerocesserom aocsro oes a cr .n vvo, cr arra.e reeas ncranscr reerreo asre-cr orre- no an c nere-cr arra are reconze as-encoeroensaare nucece aacve wee rreeneasaseroraenns a mnau crre crorsm. a s rseomnsuceeroorrmo ceavae o a e cr moecueuese cr oeare nucec ace.., aorenscr .nvan aacera ce aneas nuceaseroens ce en nucec ac e ssemsncue meansorroceasvseneeare nucec ac. a mu-roen comex cae -assocae core-cr arraancue comrse o suunsas,,, an.easca mexor anvraeense nucec acrou rzaon oear e-cr comex reconzeseare comex recrusease e nucec ac w cr.eoun nuceoroen e scasseemnsuceaseoacae ceavae oare nucec ac.racr rzeso a cr reeancnuee arrans-acvan racr .eo ceaveere-cr a e-cr arra an recrus enoenousase as.e cr-racrrra.-aesrac cromecxr reco cnomzesex cean aasrsoeca neu wcec a n aucceaser,o e.., rcz aacon ac ovaeesareea nsuc neucce aacse. wora c u nuce rre nuc.ec arcz caeoavna oe.e cr oeare ^ . asa ceaeveere-cr ssem arsranc wueeee o s ouneer oarm more o enoronoucmeaesreasse-e..e, roens. s,s anes wc are sssemans coommoroseu aes oereans seene osnas-ee-enes,ncu snsemss, e ssems are casseno severaeren sues,ncun, e. -,-,-,-,-,-,-,-,-,-,- an-.ee, e..,aaro.., a.ev.croo.,,:- anausc e a.cence,, :-va e. a. - sue encoeseasaroenormernown as .asaas auve nuceaseomanasomooousoe resecveoman oas -e omanasresennasroens.e ssems c uacse nucease oraren oeaso aare se, o an comrse a sne cr sucen aso aare se r a cr-racr uearoraren oe nown asue e. sn.oneu ecneaosneusc reeasueres c aa naucee oc aocrmanr ae rnon nuucceeocr aocen come aso w one or moreue s.n some emomens,e enonuceas x enonucease an com es ae n some em rsesracr an cr. aeno.,,.roem,e ans, cans co smeecrsee,nrome oree erm oo′moen′,s a sesaccrere exennso.. a sacer, a mnmum reea, a sneue connecor, a mn n,racr seuence, an aracr exenson.n s mumracr, a′ ac can comrse, aracr exenson, a′racrome snesaunecnecse,, a a n mucnemcu amcr-acaren nucec ue connecor, a mnmum reea, a sacer, an a r, a sne n some emomens,eue nuc sacer exensonn an orer. sneue .e comrsec ac-aren nucec ac can comrse a rzeoeare nu es a sacer seuence wc cane enneereo comrsn a s cec ac seuence.e urer comrses a reea a s euencea canrzeo aracr seuence. ona, canave conraceure exoe snason an o aracr.e exenson. reeease an eemeenrsac crannc seue eemensa can orm aase-are,oue-srane sr uence cannerac,o enonuceaseoe . ucure.e srucure canacaenn oe ^ . ′ o arsn suomeex e wmocm coemnsr,sees s an reeonu oerza coomnresewse ae sna ace mr sneuenceocae reea an mnmumracr seuence.ersuex canenerr mum ueacaes recrumen oe enonuceaseoe .euue ce aue.e rs sem comrsn aner connecne mnmum reeaaneoowe aracr seuence.easare nuceoe ae′ en oers ane mnmumo a seconner connecnersuexo a m uex cane connece one or more -racr.e m-racr can comrse n aonaarns. srucure.e soomuee emuoemens, coemrses a s caacner c eoxmenrssoen, a a soaucere, a mune nucec ac reea, a mnmumracr seuence, a′racr seuence, mum oueue oes noncuee sneue conne an aracr exenson.e reea seuence comrses a′ reea scor.nsea,e mnmum seuence comrses a′racr seu euence ane mnmumracr mnmum reea ane mnmeunmce arneoueue s canrze vae resunes some emomerns,-sa o ac rnr enn seeereuencue.e comrsn one or more seec c .n some emomens, s or userome emomensescren..aeno.,,.re, some c srucuere wa ancuees an acva sns reemo mna comeo csoem oose an u oewrou sraenxs reoromn,n a seconar reon, aue, aaren reon, a nexus, an one o aoweruexmmeaeownsream o aare r morearns. nuceoe seuence .er c mae a snne reon ma comrse varousroorons o an comrsn aaren reon, an an acuvaen re or use w ae ssem an ueruex reon, auson reon, aue, a neoxnus c,o amno osnee o or m anore aaorwenrs.u neuxc reeoone, seuen acnemme.aoer examownes,reaemar oe anar reeonn m reao cnom marse comrs oe varousroorons o , an an acvan reon ma comrse or a mxure r a mxure o an n some emomens,e enonucease useonser o an exo an . consrucnoeenome o a ces a re enousroen exresson enonucease. srcon enonucease, e.., ae resrcon ^ . consrucnno somee emenoomeme ons a, cee enson auc caeaase useonser an exoenousroen exresson caacanacveas orasa c canacve enonuceasee.., enonuceaseo.se onuaeoe ceavaeoman oe resrconoo nuceasemroeve es..,e suecnecr,. o., een aom.,e mo.causoonn, oa cuaraeocaecnnoaocv,eas –. , consruc senu seonmcee enmooemeennomse oe e anonucease useonser an exoenousroen exresson n suc emomens ces a zncner nucease , or a-ouson. reco ,eare seuences aou-aseson an comrses aar o nuceonoens soenu.enceees, e. ea.c.,m-. nu.c,e eo a.e,son,. searrae resrc ao sna ecner,m wcs-usonsooom ceea ev es:ncner n s maoeommeanns,,reo ecnoanucceaase ucseo.nser a:n exo–enous. consrucnoeenome o a ces aranscro roen exresson a -ouson.n suc emom n acvaor-e eecor nucease , or an comrses a ens,eare seuences aou- nuceoeson sac ar o reconon seuences, eac-aseson, searae a sraner, wreacs-aseson.ee e..,rsan. e a., aren oue- ns s wome em eoemceor nuceases,enecs. :–. aa c ns,ers ceavae sene ceuarenomes-ce receor aoseosnsaennn ene ane ceavaesruse ene.sruon o e s ae anen secc oemmune cesuc as ce.n some emomens, survva annns a se corme vea en omo noe omeeanves,e secoocnus. an suseuenenomc mocaons reaeo sus enneeremmune ces.n some emomens,e secon an or reeuuaenor ceaevnaese se seecsenroem ceeuarrouen coomnesncue one or moremmune ceconenes , ce, , , , ,sn, o an, -, ,m, , secon .n some emomens,e eanvea.e sene ceuarenomese ene ane ceavaesruseocae on croemosomeen,eonso acrame on. c.rommousoamneeous, couns o arme..ocues aneenes ^ . eomcuso mmaens r,esuernans aocaaaonnces aranasaonccaeon recerwoceaenr canromosomes an.n some excanears oeron ars.n some e socaon cromosomes an ure. momens,eransocaons .. ca n s coomme ermseso anme enxs,e exoenous nucec acs a -exresson consruc.ransmemraneo raceuaroman comrsn an anennn reon, a smuaoromamnsa.nn a snom oene em or moormeennrsa,ceeuaro amsaon cso smucr asses ac avaonomans an co- emomens,e aso comrses aeaereerecn neoman.n some n exemar s sownnure: “”seeαroomoee cr,e “ memrane. eaer seuence, “ ”se an- scvseecoson, e a., onsa”se an caraceraon o auncona secc sne canvramenor rucon neaeeuaema anmoma,o.mmuno.,: , “ ”s anmmunoera os aransmemraneoman, “-”s a co-smuaonoma eoman, “ ” acvaonoman, “ ”s aoaenaon sna.en, “ exrze”ssse zea ane omoo arms“ ” comrsn reons caae orznno s consrucs e ocus.eomoo armsacaeomooous recomnao euencesn oe -exressn consrucno a cune ocus.nn resunnnseron omoo arms- asearson.n some emom some emomens, eac arson. ens,eomoo arms ase ..aeneo.re,sen,nve.none mnveoovess a amn cocaonomerase can reacon , see reacon areescren rae mosonsorerormn aomerase can aer or roocos:ueesnns e a., ,caemcress,aneo,a. a , .e amcaon uzeso aneos an caonsnns e a.,caemcress, e cmomens,ormers areare-se ucscreamrmerrms,er.e a.n,r amoerwsn csormeamrsnrme ar. senu seonmcee aorememcaenareoneearee seousenocne a osesecr comeeemreennareowo.rone ooreers oorw aven amcon rmers useor amcaon.n some emomens, o ar an reverse rmerars weren aeas oner ne or more rouns o amcaon uze rmernn ses areresennmeer’-so ar uonnve“rsaa”r omer.ena sroem-es eemcocmremnesr,s u unseversoar ^ . aroro rnoaun sesu oen acme eemceanosn.noe’ u-nave.rsna somrmee ers anare-seccrmers canave uzen oonaownsream ana momens,e aona eemens are roucs. ss ses suc as caure or seuencn oe amcaon seca suea meeoorse ano cmomeraosse coansn o reaceonnv aerenoenrmnovsoavee ao nucmec acomerase.oea orea-ressan an rean sucen en erasesa are saeemerauresoreme necess zmac acv wen exoseo eevae .n some emomens,earooow eneceernmaousraaoen oomoueraes-essr caannee n uusece:cro amcsurn secese..,.avus,. ruer,.ermous,.aceus ermus searoermous,ermooa mar ,. ruens,. auacus,acus omerase,ermoco ma,eanoermuservus, omerase, oro ccus s.erees-,,, ,rococcusurosus,. ass,. .onraarsus a,n.oerramso,coc.cus s,.. s-.o,m.e sra.ses.-e,rmoco,ccuus,.oraorsonarus,. , ,rococcusurosusu,enan: , ,roceonc,cusenan: rococcus- eeen,enan: ,ermococcusoaara woes, ,enan: , ermococcu enss ,ermoc s s. sran x,enan: , ,occeursoorronaruso,: ,uoous soaarcusenan: enan: ,roacuuumm aereornxuenan: ,rcaeousuus enan: , ,ermmococecnusan:ereem,rocum occuum ,ermococcusumcoansenan: , ena,n:ermococcu, roermasenan: ,ermoco s ermococcu ccus s. enan: , ,s sr.ococ-cuse ansasn:e ,ermococcus s. enan: ,rococcusoros nean: ,rococcuscovoransenan: ,rococcus s. ennaann:: ,,errmococcus s. enan: .,ermococcus enan: ,ococecrumsocaoccusenan: ermococcusaross,ermococcus cee ccus areans, enan: ,ermococcus renan: .,ermococcusrounus ermococcus onnurneus ,u socuouse ancaonca:arum,u.o,ouesrmoocoocacus,ororeauccueunms, ^ . caenoanns:,roa,cueusmuosroacnocccuusm seceensan:,esuurococ,cus,earnooococucsu,sraonnasc aoermus,ucansaea,eanococcusenan: an oer arcum, omerases, suc asenan caea n some emomens,e, useer, . no aare nucec acsres ensuaave,.e.,eermnes weer or o sarnare nucec a enn a same wouuanne amounor co numersuanave,.e.,eermcne msoeecureessenncee a snamee. amno soumne eomr coom neunms,ere o sa ursenearren nucec ac moecuesne same.n some emomens,e useeren e r, see..,,, an..,,.s meo uzesuorescens reae-ecmoe reorse numer o amcaon cces reureo aceve areso am n an a mxure.n some emomens,euorescen emsso oun o amconn resusromncororaon o annercaane neecene course o r n some emomens,euorescen e no new sneszeoue-srane . ceavae o auorescena mssoneecene course o r resusrom omerase, ane ceavae seeroe e’-’ nucease acv oe eonan uoroore a earaeseonoruoroorerom an acceoruencer n soowmen em meoasumraenes,uoerescenc ueso occur. meo comrsnaronne sameenoerearnsons, eaaca.r a s a reacon camer were aesace.n eacaron coonecomn a mcro nucec ac moecues.acaron comrse nans one or zeroare omerase, nuceoserosaes, c s am2+caon2+reaensncun oona,aeeeeconro o-acorse.., orn, exenaermers, an comnn sameroe es.e amcaon reaens are aeoearonse.., oeorme reacon s w reaenroes or mxe we sameroroaronn aarons conan mxurenoarons. a caneerorme as en-on so anarons conannn n aon ameeccoan ree armesoeunn oar amonsc wone rree nreoseanrea nrucoenc ac wasresen, nucec ac wasresen.ounne numer oraronsn s wereeare oe numer orare nucec acsne o eac caeorrouces an esmae reerreo ans somea emrooemens, orearr .onsna e ar s ream o weaseer. c-ann-oeorromees.uocwn aan aueouss ^ . s arseoamo own sa somae ono-e ancuancsuoanenor woecs araeroromnen.n somee. e.m,uornae rocaron oso mnameer.n some emomens,e encas omens,eroes are emomens,earons are mcroues, uan osermosae.n some aronncues mcrowes or nanowes.n some emomens, eacare nuc a so suor, e.., aeasee..,,.n some emomens,e or more amec accanosnermeersar areo anasc aeacee sooe s suoor su wore.ne so smame eme noucmeecn as, one resenn souon wnearon.n some emomens,e aacmens cs cemcarousno nucec acse..,on, a sne nuc vancororan comemenar cemcarousoeeae.., s ec ac sran an conuan sran. reavn, a comemenar nucec ac sueceon seommee ermomens,eroes are arranen a monoaer ane monoaers emomens,eeaure ccno enaee rocessnseeroes.n some rome mo econ asoaesacene monoaereecneuorescence emeransocaonnno s saoeme wer. ee enmwoo smeesnns,eeennovmene oon ensn aeere assaoreecn cromosoma ses are ceavae sesor seuenc ces.n some emomens,ewo ss maeocae oneren cesr-osmeocsomce nsu.ceaesesra unsseocnaoenno mmae enen ae reercnr.ocae ceavaeransocaon,.e.,e ceave cromosome excane ae aance arms. as onear eac oe cromosome oonuceones so omre “r emmeros.”mnens,e assanvoves seuence-seccrmer w an oona some emomens, one or more ses o amcaonrmers aesn ensureseaec anon amroceonarse aeneernaoemc on seuencee nreaanrs eoaccaoransocaonuncon.uc amcon weeneraerom a w-eenome orro nasaenace, an no co oe same cromosomen m annac cromosomee.., a secon even. e ceuarenomeaas noaenarneransocaon .n som.e emomens,eransocaons aaance recrocaransocaon ene on crom.osomseran.socasorannsnovcoavoesn waesscovereene one cnvreonmoorssomnee aennomee ^ . o se ecncneere cesa ws eenreooe ocus ane ocus werearee seuence- n one emomnuenceases. crz ,envenon comrses armerar werenersrmer se esoe ene neare se o ceavae e enonucease ane eon’-ernme orerrzmeesrosee cosesenoe neaer ceea svaee o s cee.avna oee enonucease. urer comrses a seconrme ne emomen,envenon e se o ceava rar werenersrmerrzesoe ene near ene n seararnse r seea oe ve ceoaveaeermerse enonuce eanseo annuceaese s,ecuonermrmerersrrzeszeoo oeose n emomen oersrmerar. o s wereroe-aseeecons use,e urerncues aroe seconuccaeoer or “zerooe.”e ameceosnnro anucerzaeornm sere oar. areoe reuresaeroe rze eeroe seuence oroe seuence roe maeesneo enonucease a aocaonewee neare ceavae seore n some emomens,e neneersmer ane ceavae se. suc as a cme e se onseron o an exoenous seuence anroes mar ecx asnorene reeccenore succ eexssruessnon consruc.n suc an emomen,rmers ocus.ecause seron oe exoenous nucec acnoe se sucrmers anroes ma rzeoe ene neare ceavae e usoeroeeece e cnroomnuocsoemasae,raens moceaeone. reuremensorermers anroesa can .e rmers anro n exame o suc an assas sownnure -seccrm es useoeecnseronnoe ocus areare we rmern some: emers .no.mens,e rmer :sare we roe some emomens, : an : are use we n some emo:. rmer :.n sommeen ems,oemens,rmer :sare we roe : an : are use we rznno so am ceon emroo: num. ceencs a,ce seue anscsea.nu sromeren emcuoesm aenasr, oe am conroca nuocnercm acers ^ . s eemuoencmeenss a,eeno cmoncro se seueunecneceresse an snneo w e ces an enneere ces.n some enome anwce -co seuence,.e.,sresen onceneaos neoenome oe ce.n some emomens,e conro seuence somoeca eem oon amenesr,ene cromos roeamceornomureerns ceormon ceore exresson consruc.n seuence. rses aroerznoe conro sne reacnon some emomens,wo or more amcaons occur smuaneousn a , an ane voume, e..,wo or more oe eecnuson,e eecnuson n somee emeconmeen conroenomc seuence. assa comares a s s,e serorme asuanave .euanave e c naenerae eransocae cromosomeso a snaeneraeneon cerou naurceec ac amcaonoeermne co numer oeransocae cromosomes conons.nn s aomnome. mecaon conro cane useo conroor sasacor reaens an c emomens, an amcaon conros aasm comrsn aeas one room o onee oraroe se oueunscoen.n so anme eumsoonmesnees,eu corenro.asmncues amconserve asm asoncuese conro nuc n some emomens,e conro uson an ec ac a a: co rao we amconserverom n suson. ces are w-omee c eemso wmereen no, a e connroa cseauaern samace.ens s use.n some emomens, conro ces enneereosrue en ome emomens, conro ces are ces”. euacn annseronsnoe ocus“ oreecnn soemea eamnceo rmecenrso,cae cnrvoemnosoonmsa a rreaancsoocna monxunreeor ceeruoarrmen a assa emomens,eransocaons ...e reacon nome.n some onermerar wereersrmerrzes mxure comrses aeas rzeso a seuence.n so o a seuence, ane seconrmer ars, e me emomens,e reacon mxure comrseswormer rzacnoav ane sersuencrem,erunre aznnaroa ae orena soenu wence,e an oerer semcoenrarr.mer ^ . senue snocmee, emeormmeern cso,nses rneac esosnen maxure o comrses ar:s,r amner a srzno a rzno a seuence,ermer conssn essena econrmer n some emomens,e reacon mxure comrses o arsr:. seuence,ermer conssn essena o :, amnerrzno a rzno a seuence,ermer conssn essen a seconrmer n some emomens,e reacon mxure comrases o aae :. roe maeesneorzeo eer a seuence o aeeeconroe.e emomens,eroe consss essena o seuence.n some consss esse :.n some emomens,eroe nn soame o :. rmersrzn eom a coonmroen nsu,ceec r aecac soenu menxceu arenu arreoren ccauaese aar o amcaon nucec ac s oeecne conro nuc ne soumenec eem.omens, a conro reacon mxurencues an amcaon conro oeeco acow.nn s soemuee enmceso: anm aemns,coen coenrvroe nurocmec acs aasm comrsn aeas one , an. an amconerverom a conro nucec ac.usno sonme, e amn amomceonns,ereve conrroomeunseons e.., nucecn ac someso emaeormomen ws, a- conero ce resacon mxurencues conro same nucec ac, emomenns, sosmoea eemomenrso,mee nu ecnec or ne aecrese-scr cees.eren are seuence.n some emomens, amconsre cerearaons seuence.n some n one are accornoe meosescreeren are seuence. eerm "exen oera a nounmeeru oenc senuencn"e acs unsoeoeerse onr s reeeuencn assas cane uze. aowor massvearae seuencn o c rso seuencn meosa moecues. ona ame moecues an o sne nucec ac scoseereon-mn exames o seuence assasa are suaeor use we meos ,nncue nanoore seuencn..a.u.os. , , erma cce seuencnear asn e a.,oecnu,esa,ne:r se-uencn, ca, soar- arraase s seeuueennccnn, ^ . sucmm aser mmaanrx e-a ass.,seeoasesroes.oreonoo.n,z:ao-nme-o-, seue mncan w mass secromer u e a.,aureoec.,:- , seuencnss secrrozmaeornr - e a.,aureoec.,:- , an meos,ncunu nomeo seuemanac snesse..,e™,e™, orenomenazer, eac avaaero ncn seuencn aone.., ™,eecnooes,on semconuco mumna, onorren™,eecnooes, an seuencne. r seuencne.., ommerca avaae seuencnnsrumensn.,accoscences. aormsrom merxnc.unnvae,a. cue seuencn--rzaon umnaoexaaneo,a. anec , seuencn--snessaormsrom aonaormrom eo ososcencesamre,ass., seuencn--ncue,u are nom ssemsoser,a..er seuencnnsrumens ass., an nanoore-aeseo,nseruomnenosrreronmecxnooroanroomoreeremconosoerescenxorc,aam, n some emomens,envenon uzes an aaor aeo one oro e. nucec ac or nucec ac sran.aors o varous saes an ns o a see e.., e oneruar, , un acnons arenown.nn seom ar emomens,euncon o an aaorsonrouceesre eemensno e aaor-orne eemensncue aeas one o nuce a nucec ac.e aon-ena c acarcoe,rmernn se or a some emomen e a s nae. sor maeoue-srane,ara sne srane or sne srane.n oue- , a-sae, aarn aaor or a sem-oo aaors use werene escresraneeoron oe aaorsaeoeoue srane nucec acorme as seuences.nre sn no.m oeer em emoommenens,s,ee a aaaoorr m mooeeccuues aren vro snesze arca occurrn seuences.ne oer es aren vro snesze naura- occurrn mo emomens,e aaor moecues aresoae naura eecuoeus oer-ssorae non naura-occurrn moecues. overans orun ens.ann seom oer emaroamenosu,ee-sroaunee-s araaneor oon mucaeo comer casneauvne e enmsoom wencs, aeuunn-e enneaon u cnanereoe as-eano waeere a a sunn-eene nuc aeoaoer.s an oeeor ^ . ene eroouna ecnsaoe maaco ann aeaweoenreseneo anave ae s anaeor. nuceoommeer ecxaen anvaraomeesuonr errmuns asaoorceaeonuenncue c rarre or erre an aeaae maecnouons,easanon,.n some emomens,e aaor n some emome snesa,raeenrovm excess aaors an unae . emomens,e meo oeecn enonncuese use o aarcoe.n some acrocesse ase eenec mocaonsncues seuencn.e nucec mo screerens sueceo seuencnreera, massvearae sne reuecreuse a s seeuaernacene.ven oazarnconnvoura sa mmoeecueensc maaossnv aen erarroae seuencn ca moecuararcoes suc asescren..ae r correcon.e use o ,,, an,,. unue moecuararcoens aos.e,o ea,,,,,,,,o mar moecue ansroene..,e orna m c moecueoe seuence e unue moecuararcoe as mueoecue ans amconsenerae .are moecuesne same an e usesncun counne numer o orna meooruana rror correconewman,., e a., n urasensve o:. n crcuanumor wroaaen coverae,aureecne correcon.nn smo. em ene erm. eoromenens o, u an snue meoareceua mroaerccuoeess mars are useor seuencn error orms aarcoeam. varaonne seuence n e we samearcoe an amsscare as an areac.arcoes o sare a memers oearcoeareuancaon, ase enre can asoe useorosonaeucaon an ewman,., e a., nam reresens a sne moecuene orna same crcuanumor ,a erae a error suressonormroveeecon o arcoearne so nmuec eemc aocmur cee onmso oec rseseno onno ev oern moon: r,ee. ar acoaoraeo one oro ens oe e a or a muex same or useoeesn usen seuencn.arcoe can sames are mxemuexe.earcoe ma a e source oe same were n some emomens, a snearcoes us soe a comnaon o a an an. eacarcoe comrses areene se e aso an .n some emomens, ranom seuence.n some emomensue once.ennv oeneorn e,meoamrceonse,s areeaercwoeeen co amourse-s a ^ . aasrceoseosn ar seo aaeeow aeeunman anenomc samerene. ana soomrse, e eamco wme ans,eerenar oenca e reacon canen excess oe numer o moecueso numer o sn wou reconzeae numer oarcoesee eaee.erson o ornar s exece numer o unueare moec ns one comex oe same.e., arcoesor eac exe ues an woue aeo creae a suae numer o emomenns, s aomnen emrm soen usem.enos,eeerm seneu aen cconnsen sseus senvuoves seuence ann.n some e.., auraavne same unue m encerom aura o seuences, cane aeo eac moecueror oecuar .e moecuars aarcoea amcaon se.n som o seuencn or amcaon sesncue,roroe mar, a caneere esmomens, a sresenne’-oron oe rmer. comounarcoe.n oeenne’-en oeasarcoe suunoe aeoe o ens oear r emomens, a sresenn an aaor ans aeo one or seuences an saovmnee e am nuc noe enmc ce a ancs, a. coanso eenn ses.usue sneceuenaceseermnerom aura o erverome same orna moecuero vn anenca areresumeoo emnae aracs,.e., va u amcaon.n oer emomens, s use aarcuar .uc arracaso rnessu exnsnromner eorreonrs o or s ae sunence mno eercrourse ccanareac eermze usn s. nae uannn re saomvee e nmumoermse ons s,euee nnucems wer o e eaacc seue anmcoenne same caneuane muex same .ac reresens a sne moecueouaonavne same counneren s assocae w eac seuence va ene orna same an seuence varanne orna sa ran caneermneeracon o eacne ar we aeoeme, were a moecues saree same.erson se consensus seuence.n somee ermmnoemeen nus,meer re oev saenue nnucme reerass r neeacessaroeermne a e” necessaror an accuraeuanave resu ser“seuence - reas .n some emomens,eesrees eaancener re socmre. o ecma corommeonsos,maernavnesnocoanosn aneo creeuraorrmennom ae.n so asmsae emoroemecenns, ^ . remrearnsocraoznesso a . seue.nce., anee co smecornsesr ameears onrezremserar wereers n some emomens,e reacon mxure com o a seuence. rmerrzno a seuence rseswormerars, eacavners une anarae ore , ane seconrmerrzno a seuence, n some emonmaeonns w,ee c oomerrsrems aenrar. seuence,ermer conssn ess auo oersrmerrzno a rmerrzno a seuence,eenra o :, an an auo oe secon n some emomens,e comrmseer conssn essena o :. seuence,ermer conssn e s an auo oersrmerrzno a rmerrzno a seuencessena o :, an an auo oe secon n some emomens,e, ceomrmrer conssn essena o :. esneorzeo eer a seses aaeeeeconroe.eroe mae eroe consss e uence o a seuence.n some emomens, essena o ssena o :.n some emomens,eroe consss rznno s ao cmoenr eom no:. ucemcen acs, seeuenceu arner ann acuuoes o a aaror oe c amcaonrmers conro nucec aae oeecne nucec acn.n so a smc ome s e eemu emoenc ome. menens,s a,e counrroer nuncceuce asc an as auoasm o an co ammrsncaon conro oeoown seuences: an amconerveromus aeas one , an an amconerverom a on, an amconerveromuson . conro nucec ac.n some emomens,e conroenes erormnn a soomem eemrasoe cmaenns r,eaceon, suurcer as comescrrses amcaonuers an reaensor ,caemcress,aneo,a. aaee e..,n raeesnns e a., an caonsnns e a.,caemcres r or roocos:ueoeos acs amne som ase eemscroemenerse,n.ee seurs, ueenrc cno,m rrse.s reaensor seuencne nucec aaors,arcoe aa eaensncue one or more oarcoes, reensornermeae nourcs,e sec aucencnurrcameorns, s uenvs.ersarmers, an nucec acurcaon ^ . a xame.eermnne secc oeuson anusonransocaon nssa ress. u exnamroem ae recroc aass caroareeewoosseusonsuson anuson,ure a conro sne-coene was usemo asso rmeaeransocaoneween an oc. , a’-oron oe ocus an a’-reonrceonn o a muexo assa.euson assaaree ’-oron oe ocus an a’-oron oe occuuss..euurseon. a csosaaree a or eac assa an was useo assess secc.ac conroasm conanenreoaasmransocaonuson seuence ane amcon seuencen a: rao.onroaasnce was exrace usnuceonasm,nora m asm wasueo varous sm acere-ae.e euson amcaonr comnecresnraons wn assa rane usn. uronc souon. ’-en.eroe wases were : ’-en an : uson amca neor a secc seuence :.e as’-seenm.e aeccororen woan osreasme enrse we anorre aroes we se:cc seuen’-ceen an :.:e was o re creae usneuomaeroeeneraor usn-ae,eerrcmuaes c,ca.n. coen waserormen a oucermacero-ancae oma secc oe monusnexa assea..e resu cos aerseo coes aroacn ae.’- sownnae. eaen comonens n ue concaenraon rm rmxorroeso x rmeerrrrooeex reerence assax n n . n- n maesrconnzme co unessu ^ . nazeae.ermocc mecvaoncnesroeem °°amseae m:emn enauraonc: nnznemaxen : oeeacvaon n:ne a ae.aancer umsone are ansocaon assa secc coonces,. crcoeees osves eaves ao. usoncouson ,. unsroon ,. . - conro ,. ro -e csroa -nn asrcocoecaeenoonm-scecroce.roe, oes - ---nosverrooeess. ao: rao oeraanvseocraooen amconconro amconosveroesne same nxams eexam.eeecenaan acsesar oansxoacmaoens weawse aeneo e ann rouc canae “-”enneereumanonor-erve ce neere - ces e ocus an asrue ene.onro cesncues w ces we nonsereno sneroes oe c eeavee ces ano ccounsro“ ces were” m,a annua wcur-eew coe vsenroomrse a sameononr.serweo was exrace usneeasoo ssuecue aen,anenca,.aenomc n .reacon was usen . was .. e nu erorme asescrenxame. comareoe nummeerr o o aumsoncons amromcones c,o annroe numeerno omcu sseonuence amoecon as ra waos. ^ .ranesoacraeonos a croenroe s ruamo osra mnusocaeonercenaoesero amraunssooncao anneursceonna.e.ee roa are sownn esus amesaaee..ransocaonsusnon- an us- ces -- on oar.ansocaons --- - . ..-- .-- - . .. ...---- . .. ....--.-- - .. ...-- . ..-- - . ..--- .. ....we aareneo oenenv seneonnasee aerneascr venea w reerenceo secc exames, osnvenon.us,e scoe oenarous mocaons cane mae wne scoe eren,u e camsreseneeow.venon sou noeme e examesescre ^ . : rmer ’-en: rmer ’-en: roe ’-en: rmer ’-en: rmer ’-en: roe ’-en ^

Claims

.e.am a m: eneomoe o o ane eencnneeeresence o aaance recrocaransocaon .:.n c woneraecnne a sarsm ame comre car msonnam r omna mee oa rrn s m ce coare, a ene m eneeeroe c coems wrsn a:mcaonrmers, ane secon amcao orzno a seuencene ene, an a nrmers caae orzno a seuencene ene, wereorermo nnm e aenra re as sme, an ence ocra aosnence oeoe ammerasceon csann recaacvoen oeroeserouce an amcon, ran nce or asence oe .ues or mo eseca cenoon ce o. cam, wereneresence or asence oe amconseece eecn .e meo o o c aaamee, wreoreen.eransocaonseween a sene ene ,ee m seecooe ane . ne n o. am camca, wonerremners amcaonrmer consss essena o : es er consss essena o :, aneroe consss .s ,een e m sea ecoo o n o am camca, w: oner. remners amcaonrmer consss essena o : ess er consss essena o :, aneroe consss .eeen menea omo o e o o w cam,eu:r.er comrsn amn a conro nucec ac seuenceresenn e conro nuce ces anneenome o enneere ces aneecneresence o .e c ac w a conroroe. rans moecaoon o cam,u.r:er. com anrsne cuoomrarneuorescence oeroeeecne ac,ereeermnne numer oransoecsaceonncse oeroe.:eec.n amoen co enro nucec ce nneere .es m.eo o cam, werene sa . . reacon m.:xur.eorn aeeecnonme oe anre esnencnee oe aaance recrocaransocaon comrsn: re mammaan ce,e reacon mxure ^. nucrecz acnsos ao saeeuenrcoemn onee or more enennee,e arne cee sse,ers amcaonrmer caae o r con amcaonrmer caae o.e reazcnono : m a ax se nuruee onc e se ce can onme a,m werecanene one. rrmse arm conscsas eosnsenrmer consss essena o.e reacon mxure o cam, wereners amcaonrmae o :. : ane secon amcaonrmer conss r consss essena o.e reacon mxure o cam,urer comrsn os essena o :. seuence conssn essena o : ann ae s oeru meonrcee caonesesnro eess seenecae orom a.rem reera: sc. caona mex our aem o canm a, counrroer n coumcercs anc a serusen ance serecson conro amcaon ces anneenome o enneere ces, an enneenome o w e oe conro nucec ac. a conroroe caaeeecn amcaon.enomoer oe aenc ennneeerere msence o aaance recrocaransocaon .:.n a ers ammaan ce,e comrsn: se amcaonrmer caae orzno a seuencene ene, ane.ceon a omcaonrmer caae orzno a seuencene ene. an cam, wereners amcaonrmer consss essena o :.ee o sec coanm am, wcaonrmer consss essena o :. ane seco ereners amcaonrmer consss essena o :.e o canm am,urcaeorn cormer consss essena o :. con mrsn one or moreaeeroes seecerom a seuence.essn o essena o : an a seuence conssn essena o :. amn c aa cmonro, nuurceerc c aocmr sesnuen acersre asnen sencone ceonnro amcaonrmers caae o enome o enneere ces, an a conro ome o w e ces anne nucec roe caaeeecn amcaon oe conro.oemer o ac ase c. aanmu,eursr aner c coomacrosrsn ne ncuecsseaorsoe srusaons enaes, aermosae nucec ac.e o cam,urer comrsn reaen zmac acv oeomerase. seuencn nucec acs. sorrearn seuencnrares an ^

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