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pBT375 evoAPOBEC1-BE4max-NG
  • Sequence Map
    Editing is disabled because sequence edits have been locked.
    tagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgaatcaataattatcattagttaatgccccagtaatcaagtatcgggtatatacctcaaggcgcaatgtattgaatgccatttaccgggcggaccgactggcgggttgctgggggcgggtaactgcagttattactgcatacaagggtatcattgcggttatccctgaaaggtaactCMV promoter, full20406080100120140160
    cgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcagcagttacccacctcataaatgccatttgacgggtgaaccgtcatgtagttcacatagtatacggttcatgcgggggataactgcagttactgccatttaccgggcggaccgtaatacgggtcatgtactggaataccctgaaaggatgaaccgtcatgtagatgcataatcagtNdeISnaBICMV promoter, full180200220240260280300320340
    tcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatagcgataatggtaccactacgccaaaaccgtcatgtagttacccgcacctatcgccaaactgagtgcccctaaaggttcagaggtggggtaactgcagttaccctcaaacaaaaccgtggttttagttgccctgaaaggttttacagcattgttgaggcggggtaactgcgtttaNcoICspCICMV promoter, fullCMV promoter360380400420440460480500520
    gggcggtaggcgtgtacggtgggaggtctatataagcagagctggtttagtgaaccgtcagatccgctagagatccgcggccgctaatacgactcactatagggagagccgccaccatgaaacggacagccgacggaagcgagttcgagtcaccaaagaagaagcggaaagtcagcccgccatccgcacatgccaccctccagatatattcgtctcgaccaaatcacttggcagtctaggcgatctctaggcgccggcgattatgctgagtgatatccctctcggcggtggtactttgcctgtcggctgccttcgctcaagctcagtggtttcttcttcgcctttcagtcEagINotISacIIS1M1K2R3T4A5D6G7S8E9F10E11S12P13K14K15K16R17K18V19642-698CMV promoter, fullCMV promoterBipartite SV40 NLST7 promotercore promoter540560580600620640660680700
    ttcaaagactgggcctgtcgccgtcgatccaaccctgcgccgccggattgaacctcacgagtttgaagtgttctttgacccccgggagctgagaaaggagacatgcctgctgtacgagatcaactggggaggcaggcactccatctggaggcacacctctcagaacacaaataagaagtttctgacccggacagcggcagctaggttgggacgcggcggcctaacttggagtgctcaaacttcacaagaaactgggggccctcgactctttcctctgtacggacgacatgctctagttgacccctccgtccgtgaggtagacctccgtgtggagagtcttgtgtttattcSmaIXmaITspMISK2TG4PV6AV8DP10TL12RR14RI16EP18HE20FE22VF24FD26PR28EL30RK32ET34CL36LY38EI40NW42GG44RH46SI48WR50HT52SQ54NT56NK58702-1382evoAPOBEC1, Genscript codonsdispensible, de...ve (Teng 1999)dispen...1998)720740760780800820840860
    cacgtggaggtgaacttcatcgagaagtttaccacagagcggtacttctgccccaataccagatgtagcatcacatggtttctgagctggtccccttgcggagagtgtagcagggccatcaccgagttcctgtccagatatccaaatgtgacactgtttatctacatcgccAggcgtgcacctccacttgaagtagctcttcaaatggtgtctcgccatgaagacggggttatggtctacatcgtagtgtaccaaagactcgaccaggggaacgcctctcacatcgtcccggtagtggctcaaggacaggtctataggtttacactgtgacaaatagatgtagcggTccgPmlIHV60EV62NF64IE66KF68TT70ER72YF74CP76NT78RC80SI82TW84FL86SW88SP90CG92EC94SR96AI98TE100FL102SR104YP106NV108TL110FI112YI114AR116702-1382evoAPOBEC1, Genscript codonshelixhelixbeta sheetbeta sheet8809009209409609801,0001,0201,040
    tgtaTcacctggcaaacccaaggaataggcagggcctgcgcgatctgatcagctccggcgtgaccatccagatcatgacagagcaggagtccggctactgctggcacaActtcgtgaaTtattctcctagcaacgagtcccactggcctaggtacccacacctgtgggtgcgcctacatAgtggaccgtttgggttccttatccgtcccggacgcgctagactagtcgaggccgcactggtaggtctagtactgtctcgtcctcaggccgatgacgaccgtgtTgaagcacttAataagaggatcgttgctcagggtgaccggatccatgggtgtggacacccacgcggaAleIKpnIAcc65ILY118HL120AN122PR124NR126QG128LR130DL132IS134SG136VT138IQ140IM142TE144QE146SG148YC150WH152NF154VN156YS158PS160NE162SH164WP166RY168PH170LW172VR174L702-1382evoAPOBEC1, Genscript codons117-153 linker between catalytic domainshelix?specificity loophelixLeu-...ionUS...91,0601,0801,1001,1201,1401,1601,1801,2001,220
    gtacgtgctggagctgtattgcatcatcctgggcctgcccccttgtctgaatatcctgcggagaaagcagagccagctgacctcctttacaatcgccctgcagtcttgtcactatcagaggctgccaccccacatcctgtgggccacaggcctgaagtctggcggatctAgcggacatgcacgacctcgacataacgtagtaggacccggacgggggaacagacttataggacgcctctttcgtctcggtcgactggaggaaatgttagcgggacgtcagaacagtgatagtctccgacggtggggtgtaggacacccggtgtccggacttcagaccgcctagaTcgcctStuIY176VL178EL180YC182II184LG186LP188PC190LN192IL194RR196KQ198SQ200LT202SF204TI206AL208QS210CH212YQ214RL216PP218HI220LW222AT224GL226K702-1382S1G2G3S4S5G61383-1478evoAPOBEC1, Genscript codonsLeu-rich C-terminus, implicated in homodimer formationdeletion has high activity (Teng 1999)helix at dimer interfacehelixBE4 li...odonshelixbet...et1,2401,2601,2801,3001,3201,3401,3601,3801,400
    ggaTcctctggcagcgagacaccaggaacaagcgagtcagcaacaccagagagcagtggcggcagcagcggcggcagcgacaagaagtacagcatcggcctggccatcggcaccaactctgtgggctgggccgtgatcaccgacgagtacaaggtgcccagcaagaaattcaaggcctAggagaccgtcgctctgtggtccttgttcgctcagtcgttgtggtctctcgtcaccgccgtcgtcgccgccgtcgctgttcttcatgtcgtagccggaccggtagccgtggttgagacacccgacccggcactagtggctgctcatgttccacgggtcgttctttaagttccBamHIG7S8S9G10S11E12T13P14G15T16S17E18S19A20T21P22E23S24S25G26G27S28S29G30G31S321383-1478DK2KY4SI6GL8AI10GT12NS14VG16WA18VI20TD22EY24KV26PS28KK30FK321479-5579nCas9-NGBE4 linker Genscript codons1,4201,4401,4601,4801,5001,5201,5401,560
    tgctgggcaacaccgaccggcacagcatcaagaagaacctgatcggagccctgctgttcgacagcggcgaaacagccgaggccacccggctgaagagaaccgccagaagaagatacaccagacggaagaaccggatctgctatctgcaagagatcttcagcaacgagatggccaaacgacccgttgtggctggccgtgtcgtagttcttcttggactagcctcgggacgacaagctgtcgccgctttgtcggctccggtgggccgacttctcttggcggtcttcttctatgtggtctgccttcttggcctagacgatagacgttctctagaagtcgttgctctaccggttBglIIVL34GN36TD38RH40SI42KK44NL46IG48AL50LF52DS54GE56TA58EA60TR62LK64RT66AR68RR70YT72RR74KN76RI78CY80LQ82EI84FS86NE88MA90K1479-5579nCas9-NG1,5801,6001,6201,6401,6601,6801,7001,7201,740
    ggtggacgacagcttcttccacagactggaagagtccttcctggtggaagaggataagaagcacgagcggcaccccatcttcggcaacatcgtggacgaggtggcctaccacgagaagtaccccaccatctaccacctgagaaagaaactggtggacagcaccgacaaggccgacccacctgctgtcgaagaaggtgtctgaccttctcaggaaggaccaccttctcctattcttcgtgctcgccgtggggtagaagccgttgtagcacctgctccaccggatggtgctcttcatggggtggtagatggtggactctttctttgaccacctgtcgtggctgttccggctgPflMIV92DD94SF96FH98RL100EE102SF104LV106EE108DK110KH112ER114HP116IF118GN120IV122DE124VA126YH128EK130YP132TI134YH136LR138KK140LV142DS144TD146KA148D1479-5579nCas9-NG1,7601,7801,8001,8201,8401,8601,8801,9001,920
    ctgcggctgatctatctggccctggcccacatgatcaagttccggggccacttcctgatcgagggcgacctgaaccccgacaacagcgacgtggacaagctgttcatccagctggtgcagacctacaaccagctgttcgaggaaaaccccatcaacgccagcggcgtggacgccagacgccgactagatagaccgggaccgggtgtactagttcaaggccccggtgaaggactagctcccgctggacttggggctgttgtcgctgcacctgttcgacaagtaggtcgaccacgtctggatgttggtcgacaagctccttttggggtagttgcggtcgccgcacctgcggtBmgBIL150RL152IY154LA156LA158HM160IK162FR164GH166FL168IE170GD172LN174PD176NS178DV180DK182LF184IQ186LV188QT190YN192QL194FE196EN198PI200NA202SG204VD206A1479-5579nCas9-NG1,9401,9601,9802,0002,0202,0402,0602,0802,100
    aggccatcctgtctgccagactgagcaagagcagacggctggaaaatctgatcgcccagctgcccggcgagaagaagaatggcctgttcggaaacctgattgccctgagcctgggcctgacccccaacttcaagagcaacttcgacctggccgaggatgccaaactgcagctgagtccggtaggacagacggtctgactcgttctcgtctgccgaccttttagactagcgggtcgacgggccgctcttcttcttaccggacaagcctttggactaacgggactcggacccggactgggggttgaagttctcgttgaagctggaccggctcctacggtttgacgtcgactcK208AI210LS212AR214LS216KS218RR220LE222NL224IA226QL228PG230EK232KN234GL236FG238NL240IA242LS244LG246LT248PN250FK252SN254FD256LA258ED260AK262LQ264LS2661479-5579nCas9-NG2,1202,1402,1602,1802,2002,2202,2402,260
    caaggacacctacgacgacgacctggacaacctgctggcccagatcggcgaccagtacgccgacctgtttctggccgccaagaacctgtccgacgccatcctgctgagcgacatcctgagagtgaacaccgagatcaccaaggcccccctgagcgcctctatgatcaagagatacgttcctgtggatgctgctgctggacctgttggacgaccgggtctagccgctggtcatgcggctggacaaagaccggcggttcttggacaggctgcggtaggacgactcgctgtaggactctcacttgtggctctagtggttccggggggactcgcggagatactagttctctatgKD268TY270DD272DL274DN276LL278AQ280IG282DQ284YA286DL288FL290AA292KN294LS296DA298IL300LS302DI304LR306VN308TE310IT312KA314PL316SA318SM320IK322RY3241479-5579nCas9-NG2,2802,3002,3202,3402,3602,3802,4002,4202,440
    gacgagcaccaccaggacctgaccctgctgaaagctctcgtgcggcagcagctgcctgagaagtacaaagagattttcttcgaccagagcaagaacggctacgccggctacattgacggcggagccagccaggaagagttctacaagttcatcaagcccatcctggaaaagatggctgctcgtggtggtcctggactgggacgactttcgagagcacgccgtcgtcgacggactcttcatgtttctctaaaagaagctggtctcgttcttgccgatgcggccgatgtaactgccgcctcggtcggtccttctcaagatgttcaagtagttcgggtaggaccttttctaccDE326HH328QD330LT332LL334KA336LV338RQ340QL342PE344KY346KE348IF350FD352QS354KN356GY358AG360YI362DG364GA366SQ368EE370FY372KF374IK376PI378LE380KM3821479-5579nCas9-NG2,4602,4802,5002,5202,5402,5602,5802,6002,620
    acggcaccgaggaactgctcgtgaagctgaacagagaggacctgctgcggaagcagcggaccttcgacaacggcagcatcccccaccagatccacctgggagagctgcacgccattctgcggcggcaggaagatttttacccattcctgaaggacaaccgggaaaagatcgagaatgccgtggctccttgacgagcacttcgacttgtctctcctggacgacgccttcgtcgcctggaagctgttgccgtcgtagggggtggtctaggtggaccctctcgacgtgcggtaagacgccgccgtccttctaaaaatgggtaaggacttcctgttggcccttttctagctcttDG384TE386EL388LV390KL392NR394ED396LL398RK400QR402TF404DN406GS408IP410HQ412IH414LG416EL418HA420IL422RR424QE426DF428YP430FL432KD434NR436EK438IE440K1479-5579nCas9-NG2,6402,6602,6802,7002,7202,7402,7602,7802,800
    gatcctgaccttccgcatcccctactacgtgggccctctggccaggggaaacagcagattcgcctggatgaccagaaagagcgaggaaaccatcaccccctggaacttcgaggaagtggtggacaagggcgcttccgcccagagcttcatcgagcggatgaccaacttcgataagctaggactggaaggcgtaggggatgatgcacccgggagaccggtcccctttgtcgtctaagcggacctactggtctttctcgctcctttggtagtgggggaccttgaagctccttcaccacctgttcccgcgaaggcgggtctcgaagtagctcgcctactggttgaagctattcApaIPspOMII442LT444FR446IP448YY450VG452PL454AR456GN458SR460FA462WM464TR466KS468EE470TI472TP474WN476FE478EV480VD482KG484AS486AQ488SF490IE492RM494TN496FD498K1479-5579nCas9-NG2,8202,8402,8602,8802,9002,9202,9402,960
    aacctgcccaacgagaaggtgctgcccaagcacagcctgctgtacgagtacttcaccgtgtataacgagctgaccaaagtgaaatacgtgaccgagggaatgagaaagcccgccttcctgagcggcgagcagaaaaaggccatcgtggacctgctgttcaagaccaaccggaaagttggacgggttgctcttccacgacgggttcgtgtcggacgacatgctcatgaagtggcacatattgctcgactggtttcactttatgcactggctcccttactctttcgggcggaaggactcgccgctcgtctttttccggtagcacctggacgacaagttctggttggcctttcBarIN500LP502NE504KV506LP508KH510SL512LY514EY516FT518VY520NE522LT524KV526KY528VT530EG532MR534KP536AF538LS540GE542QK544KA546IV548DL550LF552KT554NR556K1479-5579nCas9-NG2,9803,0003,0203,0403,0603,0803,1003,1203,140
    tgaccgtgaagcagctgaaagaggactacttcaagaaaatcgagtgcttcgactccgtggaaatctccggcgtggaagatcggttcaacgcctccctgggcacataccacgatctgctgaaaattatcaaggacaaggacttcctggacaatgaggaaaacgaggacattctggaactggcacttcgtcgactttctcctgatgaagttcttttagctcacgaagctgaggcacctttagaggccgcaccttctagccaagttgcggagggacccgtgtatggtgctagacgacttttaatagttcctgttcctgaaggacctgttactccttttgctcctgtaagacctPasIV558TV560KQ562LK564ED566YF568KK570IE572CF574DS576VE578IS580GV582ED584RF586NA588SL590GT592YH594DL596LK598II600KD602KD604FL606DN608EE610NE612DI614LE6161479-5579nCas9-NG3,1603,1803,2003,2203,2403,2603,2803,3003,320
    agatatcgtgctgaccctgacactgtttgaggacagagagatgatcgaggaacggctgaaaacctatgcccacctgttcgacgacaaagtgatgaagcagctgaagcggcggagatacaccggctggggcaggctgagccggaagctgatcaacggcatccgggacaagcagtcctctatagcacgactgggactgtgacaaactcctgtctctctactagctccttgccgacttttggatacgggtggacaagctgctgtttcactacttcgtcgacttcgccgcctctatgtggccgaccccgtccgactcggccttcgactagttgccgtaggccctgttcgtcaggAhdIDI618VL620TL622TL624FE626DR628EM630IE632ER634LK636TY638AH640LF642DD644KV646MK648QL650KR652RR654YT656GW658GR660LS662RK664LI666NG668IR670DK672QS6741479-5579nCas9-NG3,3403,3603,3803,4003,4203,4403,4603,4803,500
    ggcaagacaatcctggatttcctgaagtccgacggcttcgccaacagaaacttcatgcagctgatccacgacgacagcctgacctttaaagaggacatccagaaagcccaggtgtccggccagggcgatagcctgcacgagcacattgccaatctggccggcagccccgccattaccgttctgttaggacctaaaggacttcaggctgccgaagcggttgtctttgaagtacgtcgactaggtgctgctgtcggactggaaatttctcctgtaggtctttcgggtccacaggccggtcccgctatcggacgtgctcgtgtaacggttagaccggccgtcggggcggtaatAjuIGK676TI678LD680FL682KS684DG686FA688NR690NF692MQ694LI696HD698DS700LT702FK704ED706IQ708KA710QV712SG714QG716DS718LH720EH722IA724NL726AG728SP730AI7321479-5579nCas9-NG3,5203,5403,5603,5803,6003,6203,6403,660
    agaagggcatcctgcagacagtgaaggtggtggacgagctcgtgaaagtgatgggccggcacaagcccgagaacatcgtgatcgaaatggccagagagaaccagaccacccagaagggacagaagaacagccgcgagagaatgaagcggatcgaagagggcatcaaagagctgggtcttcccgtaggacgtctgtcacttccaccacctgctcgagcactttcactacccggccgtgttcgggctcttgtagcactagctttaccggtctctcttggtctggtgggtcttccctgtcttcttgtcggcgctctcttacttcgcctagcttctcccgtagtttctcgacccSacIEco53kIKK734GI736LQ738TV740KV742VD744EL746VK748VM750GR752HK754PE756NI758VI760EM762AR764EN766QT768TQ770KG772QK774NS776RE778RM780KR782IE784EG786IK788EL790G1479-5579nCas9-NG3,6803,7003,7203,7403,7603,7803,8003,8203,840
    cagccagatcctgaaagaacaccccgtggaaaacacccagctgcagaacgagaagctgtacctgtactacctgcagaatgggcgggatatgtacgtggaccaggaactggacatcaaccggctgtccgactacgatgtggaccatatcgtgcctcagagctttctgaaggacgacgtcggtctaggactttcttgtggggcaccttttgtgggtcgacgtcttgctcttcgacatggacatgatggacgtcttacccgccctatacatgcacctggtccttgacctgtagttggccgacaggctgatgctacacctggtatagcacggagtctcgaaagacttcctgctgPflMIDraIIIS792QI794LK796EH798PV800EN802TQ804LQ806NE808KL810YL812YY814LQ816NG818RD820MY822VD824QE826LD828IN830RL832SD834YD836VD838HI840VP842QS844FL846KD848D1479-5579nCas9-NG3,8603,8803,9003,9203,9403,9603,9804,0004,020
    tccatcgacaacaaggtgctgaccagaagcgacaagaaccggggcaagagcgacaacgtgccctccgaagaggtcgtgaagaagatgaagaactactggcggcagctgctgaacgccaagctgattacccagagaaagttcgacaatctgaccaaggccgagagaggcggcctgaaggtagctgttgttccacgactggtcttcgctgttcttggccccgttctcgctgttgcacgggaggcttctccagcacttcttctacttcttgatgaccgccgtcgacgacttgcggttcgactaatgggtctctttcaagctgttagactggttccggctctctccgccggactS850ID852NK854VL856TR858SD860KN862RG864KS866DN868VP870SE872EV874VK876KM878KN880YW882RQ884LL886NA888KL890IT892QR894KF896DN898LT900KA902ER904GG906L1479-5579nCas9-NG4,0404,0604,0804,1004,1204,1404,1604,1804,200
    gcgaactggataaggccggcttcatcaagagacagctggtggaaacccggcagatcacaaagcacgtggcacagatcctggactcccggatgaacactaagtacgacgagaatgacaagctgatccgggaagtgaaagtgatcaccctgaagtccaagctggtgtccgatttccgcgcttgacctattccggccgaagtagttctctgtcgaccacctttgggccgtctagtgtttcgtgcaccgtgtctaggacctgagggcctacttgtgattcatgctgctcttactgttcgactaggcccttcactttcactagtgggacttcaggttcgaccacaggctaaaggcPmlICspCIS908EL910DK912AG914FI916KR918QL920VE922TR924QI926TK928HV930AQ932IL934DS936RM938NT940KY942DE944ND946KL948IR950EV952KV954IT956LK958SK960LV962SD964FR9661479-5579nCas9-NG4,2204,2404,2604,2804,3004,3204,3404,360
    gaaggatttccagttttacaaagtgcgcgagatcaacaactaccaccacgcccacgacgcctacctgaacgccgtcgtgggaaccgccctgatcaaaaagtaccctaagctggaaagcgagttcgtgtacggcgactacaaggtgtacgacgtgcggaagatgatcgccaagagccttcctaaaggtcaaaatgtttcacgcgctctagttgttgatggtggtgcgggtgctgcggatggacttgcggcagcacccttggcgggactagtttttcatgggattcgacctttcgctcaagcacatgccgctgatgttccacatgctgcacgccttctactagcggttctcgBmgBIKD968FQ970FY972KV974RE976IN978NY980HH982AH984DA986YL988NA990VV992GT994AL996IK998KY1000PK1002LE1004SE1006FV1008YG1010DY1012KV1014YD1016VR1018KM1020IA1022KS10241479-5579nCas9-NG4,3804,4004,4204,4404,4604,4804,5004,5204,540
    gagcaggaaatcggcaaggctaccgccaagtacttcttctacagcaacatcatgaactttttcaagaccgagattaccctggccaacggcgagatccggaagcggcctctgatcgagacaaacggcgaaaccggggagatcgtgtgggataagggccgggattttgccaccgtgcctcgtcctttagccgttccgatggcggttcatgaagaagatgtcgttgtagtacttgaaaaagttctggctctaatgggaccggttgccgctctaggccttcgccggagactagctctgtttgccgctttggcccctctagcacaccctattcccggccctaaaacggtggcacgAjuIEQ1026EI1028GK1030AT1032AK1034YF1036FY1038SN1040IM1042NF1044FK1046TE1048IT1050LA1052NG1054EI1056RK1058RP1060LI1062ET1064NG1066ET1068GE1070IV1072WD1074KG1076RD1078FA1080TV10821479-5579nCas9-NG4,5604,5804,6004,6204,6404,6604,6804,7004,720
    ggaaagtgctgagcatgccccaagtgaatatcgtgaaaaagaccgaggtgcagacaggcggcttcagcaaagagtctatccggcccaagaggaacagcgataagctgatcgccagaaagaaggactgggaccctaagaagtacggcggcttcgtgagccccaccgtggcctattccctttcacgactcgtacggggttcacttatagcactttttctggctccacgtctgtccgccgaagtcgtttctcagataggccgggttctccttgtcgctattcgactagcggtctttcttcctgaccctgggattcttcatgccgccgaagcactcggggtggcaccggataagBarIKflISphIRK1084VL1086SM1088PQ1090VN1092IV1094KK1096TE1098VQ1100TG1102GF1104SK1106ES1108IR1110PK1112RN1114SD1116KL1118IA1120RK1122KD1124WD1126PK1128KY1130GG1132FV1134SP1136TV1138AY1140S1479-5579nCas9-NG4,7404,7604,7804,8004,8204,8404,8604,8804,900
    tgtgctggtggtggccaaagtggaaaagggcaagtccaagaaactgaagagtgtgaaagagctgctggggatcaccatcatggaaagaagcagcttcgagaagaatcccatcgactttctggaagccaagggctacaaagaagtgaaaaaggacctgatcatcaagctgcctaagacacgaccaccaccggtttcaccttttcccgttcaggttctttgacttctcacactttctcgacgacccctagtggtagtacctttcttcgtcgaagctcttcttagggtagctgaaagaccttcggttcccgatgtttcttcactttttcctggactagtagttcgacggattcXcmIBsaBIV1142LV1144VA1146KV1148EK1150GK1152SK1154KL1156KS1158VK1160EL1162LG1164IT1166IM1168ER1170SS1172FE1174KN1176PI1178DF1180LE1182AK1184GY1186KE1188VK1190KD1192LI1194IK1196LP1198K1479-5579nCas9-NG4,9204,9404,9604,9805,0005,0205,0405,060
    tactccctgttcgagctggaaaacggccggaagagaatgctggcctctgcccggttcctgcagaagggaaacgaactggccctgccctccaaatatgtgaacttcctgtacctggccagccactatgagaagctgaagggctcccccgaggataatgagcagaaacagctgtttgatgagggacaagctcgaccttttgccggccttctcttacgaccggagacgggccaaggacgtcttccctttgcttgaccgggacgggaggtttatacacttgaaggacatggaccggtcggtgatactcttcgacttcccgagggggctcctattactcgtctttgtcgacaaacBsmIEagIY1200SL1202FE1204LE1206NG1208RK1210RM1212LA1214SA1216RF1218LQ1220KG1222NE1224LA1226LP1228SK1230YV1232NF1234LY1236LA1238SH1240YE1242KL1244KG1246SP1248ED1250NE1252QK1254QL1256F1479-5579nCas9-NG5,0805,1005,1205,1405,1605,1805,2005,2205,240
    tggaacagcacaagcactacctggacgagatcatcgagcagatcagcgagttctccaagagagtgatcctggccgacgctaatctggacaaagtgctgtccgcctacaacaagcaccgggataagcccatcagagagcaggccgagaatatcatccacctgtttaccctgaccaaaccttgtcgtgttcgtgatggacctgctctagtagctcgtctagtcgctcaagaggttctctcactaggaccggctgcgattagacctgtttcacgacaggcggatgttgttcgtggccctattcgggtagtctctcgtccggctcttatagtaggtggacaaatgggactggttV1258EQ1260HK1262HY1264LD1266EI1268IE1270QI1272SE1274FS1276KR1278VI1280LA1282DA1284NL1286DK1288VL1290SA1292YN1294KH1296RD1298KP1300IR1302EQ1304AE1306NI1308IH1310LF1312TL1314TN13161479-5579nCas9-NG5,2605,2805,3005,3205,3405,3605,3805,4005,420
    tctgggagcccctcgggccttcaagtactttgacaccaccatcgaccggaaggtgtaccggagcaccaaagaggtgctggacgccaccctgatccaccagagcatcaccggcctgtacgagacacggatcgacctgtctcagctgggaggtgacagcggcgggagcggcgggagcagaccctcggggagcccggaagttcatgaaactgtggtggtagctggccttccacatggcctcgtggtttctccacgacctgcggtgggactaggtggtctcgtagtggccggacatgctctgtgcctagctggacagagtcgaccctccactgtcgccgccctcgccgccctcgXcmILG1318AP1320RA1322FK1324YF1326DT1328TI1330DR1332KV1334YR1336ST1338KE1340VL1342DA1344TL1346IH1348QS1350IT1352GL1354YE1356TR1358ID1360LS1362QL1364GG1366D1479-5579SG2GS4GG6S5580-6149nCas9-NG2xUGI G...codons5,4405,4605,4805,5005,5205,5405,5605,5805,600
    ggggggagcactaatctgagcgacatcattgagaaggagactgggaaacagctggtcattcaggagtccatcctgatgctgcctgaggaggtggaggaagtgatcggcaacaagccagagtctgacatcctggtgcacaccgcctacgacgagtccacagatgagaatgtgatgccccccctcgtgattagactcgctgtagtaactcttcctctgaccctttgtcgaccagtaagtcctcaggtaggactacgacggactcctccacctccttcactagccgttgttcggtctcagactgtaggaccacgtgtggcggatgctgctcaggtgtctactcttacactacgBseRIBsu36IG8GS10TN12LS14DI16IE18KE20TG22KQ24LV26IQ28ES30IL32ML34PE36EV38EE40VI42GN44KP46ES48DI50LV52HT54AY56DE58ST60DE62NV64M5580-61492xUGI Genscript codons5,6205,6405,6605,6805,7005,7205,7405,760
    tgctgacctctgacgcccccgagtataagccttgggccctggtcatccaggattctaacggcgagaataagatcaagatgctgagcggaggatccggaggatctggaggcagcaccaacctgtctgacatcatcgagaaggagacaggcaagcagctggtcatccaggagagcatacgactggagactgcgggggctcatattcggaacccgggaccagtaggtcctaagattgccgctcttattctagttctacgactcgcctcctaggcctcctagacctccgtcgtggttggacagactgtagtagctcttcctctgtccgttcgtcgaccagtaggtcctctcgtaApaIBamHIPspOMIL66LT68SD70AP72EY74KP76WA78LV80IQ82DS84NG86EN88KI90KM92LS94GG96SG98GS100GG102ST104NL106SD108II110EK112ET114GK116QL118VI120QE122SI1245580-61492xUGI Genscript codons5,7805,8005,8205,8405,8605,8805,9005,9205,940
    cctgatgctgcccgaagaagtcgaagaagtgatcggaaacaagcctgagagcgatatcctggtccataccgcctacgacgagagtaccgacgaaaatgtgatgctgctgacatccgacgccccagagtataagccctgggctctggtcatccaggattccaacggagagaacaaaggactacgacgggcttcttcagcttcttcactagcctttgttcggactctcgctataggaccaggtatggcggatgctgctctcatggctgcttttacactacgacgactgtaggctgcggggtctcatattcgggacccgagaccagtaggtcctaaggttgcctctcttgtttBaeIPasILM126LP128EE130VE132EV134IG136NK138PE140SD142IL144VH146TA148YD150ES152TD154EN156VM158LL160TS162DA164PE166YK168PW170AL172VI174QD176SN178GE180NK1825580-61492xUGI Genscript codons5,9605,9806,0006,0206,0406,0606,0806,1006,120
    atcaaaatgctgtctggcggctcaaaaagaaccgccgacggcagcgaattcgagcccaagaagaagaggaaagtctaaccggtcatcatcaccatcaccattgagtttAaacccgcTgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctcccccgttagttttacgacagaccgccgagtttttcttggcggctgccgtcgcttaagctcgggttcttcttctcctttcagattggccagtagtagtggtagtggtaactcaaaTttgggcgActagtcggagctgacacggaagatcaacggtcggtagacaacaaacggggagggggcaAgeIPmeIEcoRI*1IK184ML186SG188GS1905580-6149K1R2T3A4D5G6S7E8F9E10P11K12K13K14R15K16V176150-6200bGH PA terminatorSV40 NLS2xUGI Gen...t codonsSV40 NLS6,1406,1606,1806,2006,2206,2406,2606,2806,300
    gccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctcggaaggaactgggaccttccacggtgagggtgacaggaaaggattattttactcctttaacgtagcgtaacagactcatccacagtaagataagaccccccaccccaccccgtcctgtcgttccccctcctaacccttctgttatcgtccgtacgacccctacgccacccgagaBbsISphIbGH PA terminator6,3206,3406,3606,3806,4006,4206,4406,460
    atggcttctgaggcggaaagaaccagctggggctcgataccgtcgacctctagctagagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaagtgtaaagcctagggtgcctaatgagtgataccgaagactccgcctttcttggtcgaccccgagctatggcagctggagatcgatctcgaaccgcattagtaccagtatcgacaaaggacacactttaacaataggcgagtgttaaggtgtgttgtatgctcggccttcgtatttcacatttcggatcccacggattactcactlac promoter6,4806,5006,5206,5406,5606,5806,6006,6206,640
    gctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgcActtccgcTtcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagccgattgagtgtaattaacgcaacgcgagtgacgggcgaaaggtcagccctttggacagcacggtcgacgtaattacttagccggttgcgcgcccctctccgccaaacgcataacccgcgTgaaggcgAaggagcgagtgactgagcgacgcgagccagcaagccgacgccgctcg6,6606,6806,7006,7206,7406,7606,7806,8006,820
    ggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaccatagtcgagtgagtttccgccattatgccaataggtgtcttagtcccctattgcgtcctttcttgtacactcgttttccggtcgttttccggtccttggcatttttccggcgcaacgaccgcaaaaaggtatccgaggcggggggactgctcgtagtgtttttagctgcgagtDrdIPciIAflIIIpUC origin high copy6,8406,8606,8806,9006,9206,9406,9606,9807,000
    agtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtcagtctccaccgctttgggctgtcctgatatttctatggtccgcaaagggggaccttcgagggagcacgcgagaggacaaggctgggacggcgaatggcctatggacaggcggaaagagggaagcccttcgcaccgcgaaagagtatcgagtgcgacatccatagagtcaagccBciVIpUC origin high copy7,0207,0407,0607,0807,1007,1207,1407,160
    tgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacacatccagcaagcgaggttcgacccgacacacgtgcttggggggcaagtcgggctggcgacgcggaataggccattgatagcagaactcaggttgggccattctgtgctgaatagcggtgaccgtcgtcggtgaccattgtcctaatcgtctcgctccatacatccgccacgatgpUC origin high copy7,1807,2007,2207,2407,2607,2807,3007,3207,340
    agagttcttgaagtggtggcctaactacggctacactagaagAacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagatctcaagaacttcaccaccggattgatgccgatgtgatcttcTtgtcataaaccatagacgcgagacgacttcggtcaatggaagcctttttctcaaccatcgagaactaggccgtttgtttggtggcgaccatcgccaccaaaaaaacaaacgttcgtcgtctaatgcgcgtctpUC origin high copy7,3607,3807,4007,4207,4407,4607,4807,5007,520
    aaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacActcagtggaacgaaaactcacgttaagggAttttggtcaTgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttttttttcctagagttcttctaggaaactagaaaagatgccccagactgTgagtcaccttgcttttgagtgcaattcccTaaaaccagtActctaatagtttttcctagaagtggatctaggaaaatttaatttttacttcaaaatttagttagatttcatatatactcatttgaaDraIIIpUC origin high copyUS...57,5407,5607,5807,6007,6207,6407,6607,6807,700
    ggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgGgAcccacgcTcaccggctccagaccagactgtcaatggttacgaattagtcactccgtggatagagtcgctagacagataaagcaagtaggtatcaacggactgaggggcagcacatctattgatgctatgccctcccgaatggtagaccggggtcacgacgttactatggcgcCcTgggtgcgAgtggccgaggtctAhdIKflISacII*W286HK284IL282SA280GI278EA276IQ274RN272RE270DM268TA266QS264GT262TY260IV258VI256RS254PK252GD250PG248LA246AI244IG242RS240GR238EG236AG234S7711-8571bla beta lactamase (ampicillin/carbenicillin resistance)7,7207,7407,7607,7807,8007,8207,8407,860
    tttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgaaatagtcgttatttggtcggtcggccttcccggctcgcgtcttcaccaggacgttgaaataggcggaggtaggtcagataattaacaacggcccttcgatctcattcatcaagcggtcaattatcaaacgcgttgcaacaacggtaacgatgtccgtagcaccacagtgcgagcAclIFspIK232DA230IF228WG226AP224LA222SR220LL218PG216AV214KD212AE210MW208DI206LQ204QR202SA200LT198LL196EG194TL192LK190RL188TT186AM184AV182PM180TT178DR176E7711-8571bla beta lactamase (ampicillin/carbenicillin resistance)7,8807,9007,9207,9407,9607,9808,0008,0208,040
    tcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgagcaaaccataccgaagtaagtcgaggccaagggttgctagttccgctcaatgtactagggggtacaacacgttttttcgccaatcgaggaagccaggaggctagcaacagtcttcattcaaccggcgtcacaatagtgagtaccaataccgtcgtgacgtattaagagaatgacPvuID174NP172IA170EN168LE166PE164WR162DL160RT158VH156DG154MN152HL150FA148TL146EK144PG142GI140TT138LL136LN134AA132TN130DS128MT126IA124AS122CL120ER118VT1167711-8571bla beta lactamase (ampicillin/carbenicillin resistance)8,0608,0808,1008,1208,1408,1608,1808,2008,220
    tcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaagtacggtaggcattctacgaaaagacactgaccactcatgagttggttcagtaagactcttatcacatacgccgctggctcaacgagaacgggccgcagttatgccctattatggcgcggtgtatcgtcttgaaattttcacgagtagtaaccttttgcaagaagccccgctttAclIMG114DT112LH110KE108TV106PS104YE102VL100DN98QS96YH94IR92RG90LQ88EQ86GA84DI82RS80LV78AG76CL74LV72KF70TS68MM66PF64RE62EP60RF587711-8571bla beta lactamase (ampicillin/carbenicillin resistance)8,2408,2608,2808,3008,3208,3408,3608,3808,400
    actctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactctgagagttcctagaatggcgacaactctaggtcaagctacattgggtgagcacgtgggttgactagaagtcgtagaaaatgaaagtggtcgcaaagacccactcgtttttgtccttccgttttacggcgttttttcccttattcccgctgtgcctttacaacttatgagtatgagSE56LI54KG52SN50LD48LE46IY44GV42RA40GL38QD36EA34DK32VK30VL28TE26PH24AF22VP20LC18FA16AF14FP12IL10AV8RF6HQ4IS2M7711-8571bla beta lactamase (ampicillin/carbenicillin resistance)8,4208,4408,4608,4808,5008,5208,5408,560
    ttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtcgacggatcgggagatcgatctcccgatcccctagggtcgactcaaggaaaaagttataataacttcgtaaatagtcccaataacagagtactcgcctatgtataaacttacataaatctttttatttgtttatccccaaggcgcgtgtaaaggggcttttcacggtggactgcagctgcctagccctctagctagagggctaggggatcccagctgagClaISspIBciVIBspDISgrDIamp promoter8,5808,6008,6208,6408,6608,6808,7008,7208,740
    tcagtacaatctgctctgatgccgcatagttaagccagtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcgcgagcaaaatttaagctacaacaaggcaaggcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttcgcgatgtagtcatgttagacgagactacggcgtatcaattcggtcatagacgagggacgaacacacaacctccagcgactcatcacgcgctcgttttaaattcgatgttgttccgttccgaactggctgttaacgtacttcttagacgaatcccaatccgcaaaacgcgacgaagcgctacaMfeINruI8,7608,7808,8008,8208,8408,8608,8808,9008,920
    acgggccagatatacgcgttgacattgattattgactgcccggtctatatgcgcaactgtaactaataactgMluISpeIAflIII8,9408,960
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    8961 bpnCas9-NG Nishimasu et al. Science 2018, Genscript codonspBT195 backbone65007000750080008500500100015002000nCas9-NGUSER junction 331479-5579CMV promoter, full,NdeI,SnaBI,NcoI,CspCICMV promoter,core promoter,T7 promoter,essential bases,essential base,+14+16evoAPOBEC1, Genscript codons,beta sheet,crosslinks to apoB mRNA,+18gap for RNA binding,R154H,+14Leu-rich C-terminus, implicated in homodimer formation,+13BamHIBE4 linker Genscript codons,1383-1478BglIIPflMIBmgBIremoved SapI site,USER jct 26AflIII,PciIDrdIBciVIpUC origin high copyG>A,USER jct 25,DraIIIremoved BsaI site,AhdI,SacII,KflIUSER jct 31,FspI,AclIbla beta lactamase (ampicillin/carbenicillin resistance),7711-8571,PvuIAclIamp promoter,SspI,BciVI,SgrDI,BspDI,ClaI,MfeI,NruI,AflIII,MluI,SpeI

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    8961