                         SEQUENCE LISTING

<110>  Beijing Biocytogen Co., Ltd
 
<120>  GENETICALLY MODIFIED NON-HUMAN ANIMALS WITH KIT MUTATIONS

<130>  1

<160>  18    

<170>  PatentIn version 3.5

<210>  1
<211>  5189
<212>  DNA
<213>  Mouse

<400>  1
ggtgcacttg ggcgagagct gtagcagaga gaggagctca gagtctagcg cagccaccgc       60

gatgagaggc gctcgcggcg cctgggatct gctctgcgtc ctgttggtcc tgctccgtgg      120

ccagacagcc acgtctcagc catctgcaag tccaggggag ccgtctccgc catccatcca      180

tccagcacaa tcagagttaa tagttgaagc tggcgacacc ctcagcctga cgtgcattga      240

tcccgacttt gtcagatgga ctttcaagac ctatttcaat gaaatggttg agaataaaaa      300

aaatgaatgg atccaggaaa aagccgaggc cactcgcacg ggcacataca cgtgcagcaa      360

cagcaatggc ctcacgagtt ctatttacgt gtttgttaga gatcctgcca aacttttcct      420

ggttggcctt cccttgtttg gcaaagaaga cagcgacgcg ctggtccgct gccctctgac      480

agacccacag gtgtccaatt attccctcat cgagtgtgat gggaaatctc tccccacgga      540

cctgacgttt gtcccaaacc ccaaggctgg catcaccatc aaaaacgtga agcgcgccta      600

ccaccggctc tgtgtccgct gtgctgctca gcgtgacggt acatggctgc attctgacaa      660

attcaccctc aaagtgcggg cagccatcaa ggctatccct gttgtgtctg tgcctgaaac      720

aagtcacctc cttaagaaag gggacacatt tacggtggtg tgcaccataa aagatgtgtc      780

tacatccgtg aactccatgt ggctaaagat gaaccctcag cctcagcaca tagcccaggt      840

aaagcacaat agctggcacc ggggtgactt caattatgaa cgccaggaga cgctgactat      900

cagctcggca agagttgacg attctggagt gttcatgtgt tatgccaata atacttttgg      960

atcagcaaat gtcacaacaa ccttgaaagt agtagaaaaa ggattcatca acatctcccc     1020

tgtgaagaac actacagtat ttgtaaccga tggagaaaac gtagatttgg ttgttgaata     1080

cgaggcctac cccaaacccg agcaccagca gtggatatat atgaacagga cctcggctaa     1140

caaagggaag gattatgtca aatctgataa caaaagcaac atcagatatg tgaaccaact     1200

tcgcctgacc agattaaaag gcacagaagg aggcacttat acctttctgg tgtccaactc     1260

tgatgccagt gcttccgtga cattcaacgt ttacgtgaac acaaaaccag aaatcctgac     1320

gtacgacagg ctcataaatg gcatgctcca gtgtgtggca gagggattcc cggagcccac     1380

aatagattgg tatttttgta caggagcaga gcaaaggtgt accactcctg tctcaccagt     1440

ggacgtacag gtccagaatg tatctgtgtc accatttgga aaactggtgg ttcagagttc     1500

catagactcc agcgtcttcc ggcacaacgg cacggtggag tgtaaggcct ccaacgatgt     1560

gggcaagagt tccgccttct ttaactttgc atttaaagag caaatccagg cccacactct     1620

gttcacgccg ctgctcattg gctttgtggt tgcagctggc gcgatgggga tcattgtgat     1680

ggtgctcacc tacaaatatt tgcagaaacc catgtatgaa gtacaatgga aggttgtcga     1740

ggagataaat ggaaacaatt atgtttacat agacccgacg caacttcctt atgatcacaa     1800

atgggagttt cccagaaaca ggctgagttt tggaaagaca ttgggagctg gtgccttcgg     1860

gaaggtcgtt gaggccactg catatggctt gattaagtcg gatgctgcca tgacagttgc     1920

cgtgaagatg ctcaaaccaa gtgcccattt aacagaaaga gaggccctaa tgtcggaact     1980

gaaggtcctg agctacctgg gcaatcacat gaatattgtg aacctgcttg gcgcatgcac     2040

ggtgggaggg cccaccctgg tcattacaga atattgttgc tatggtgatc ttttgaattt     2100

tttgagaagg aagcgtgact cgtttatttt ctcaaagcaa gaagagcagg cagaagcggc     2160

actttataag aaccttctgc actcaacgga gccttcctgt gacagttcaa atgaatatat     2220

ggacatgaag cctggcgttt cctacgtggt gccaaccaag acagacaaga ggagatccgc     2280

aagaatagac tcgtacatag aaagagacgt gactcctgcc atcatggaag atgacgagct     2340

ggctctggac ctggatgatt tgctgagctt ctcctaccag gtggccaagg gcatggcgtt     2400

cctcgcctcc aagaattgta ttcacagaga tttggcagcc aggaatatcc tcctcactca     2460

cgggcggatc acaaagattt gcgatttcgg gctagccaga gacatcagga atgattcgaa     2520

ttacgtggtc aaaggaaatg cacgactgcc cgtgaagtgg atggcaccag agagcatttt     2580

cagctgcgtg tacacatttg aaagtgatgt ctggtcctat gggattttcc tctgggagct     2640

cttctcctta ggaagcagcc cctacccagg gatgccggtc gactccaagt tctacaagat     2700

gatcaaggaa ggcttccgga tggtcagccc ggagcacgcg cctgccgaaa tgtatgacgt     2760

catgaagact tgctgggacg ctgacccctt gaaaaggcca acattcaagc aggttgtcca     2820

acttattgag aagcagatct cggacagcac caagcacatt tactccaact tggcaaactg     2880

caaccccaac ccagagaacc ccgtggtggt ggaccattcc gtgagggtca actcggtggg     2940

cagcagcgcc tcttctacgc agcccctgct cgtgcacgaa gatgcctgag cagaaaccca     3000

agtccaacag gctttgctgc tgtctccgac cccgtccttc tggcttctgt gatggttact     3060

tggtttccct ttgacttgca tcctattcca gggtagcgag ttccccaccc cacctccaac     3120

cccactgtga ttccgccttt acgagcacac actttagtgg ccgatggcct tttcttttct     3180

gccatcagcc accgtcccgc tgcgaaggtc cgaactgtat gtatatattt tcccaatagc     3240

aaagtagctc ctactgtaaa cagaaggact cctcctgctt tagaggagaa gggaagggcg     3300

gggtgaaact ggatgcccag agttcttccc ccagtgctcc cctgagtgta tttgaaaagt     3360

atggccagta gttcacttga agaatagatg tagtcccatt tggccctgag agccatcctt     3420

aatgatggga gatatatgta gcaagactag aaaaggaaag ccaagccctt tgtgtagaaa     3480

gcagaccatt cttagaacag agggcaacgg ggcatcggaa gtctggtcac gctaagaaga     3540

ccgaggctga gaaggaacaa gccaggggaa gcgtgaacaa tgatgctctg ctctgggctg     3600

ccgctcgggc ttctgtacaa ctgacctggt ttctcagtac tttgctgtct gggagtagca     3660

ttggaatcaa ggcctcctcc ctagtcagcc tttgtatata ctcatctata cgttgtatgc     3720

gttcatactt tggaggaggg atttcccaca agctttcgtt tctgtgtaca gccctggcat     3780

tagacctact gtgtgtaaga atagattaag agccatacat atttgaagga aacagttaaa     3840

tgttttttgg ttgtggttgt tgttgttgtt gttttaaaga aaaaaatgta tatgctaagc     3900

acaatcttta taagacctct tagccaacat acttgctctg tctacacttc ggaacaagcc     3960

ttccatgtca gagtggcttt gcaggcagga gaactgaggc tgtttgaaaa ggttaccaca     4020

ggatggagaa aacagtgcag tcctggtttg gattctcaca tagcagggag cacaagttaa     4080

actcagcctt ttataggcac gtcccggaca tcgggccagt atctattcaa gtgtgtatgt     4140

gtgtgcatgc gtgtgtctat gcgtgtgggt gagttgtgtt gggaaacttg ccctgcatcc     4200

ctgagggtcc tccttcagga cccaagacgt aacagcttct gtcaccgctc ctgtctctcc     4260

agtttccctg catgtcgctc actgtctaga atttactcaa agccgccaca gaggcttagc     4320

ggagtgaagt gccgaaggac ctctttattt ggagtcctcc tgtatttaac aacactctta     4380

tcgtagaccc attcattaga ccttatgtaa tgctgccaat ccagggaaac agatttaaag     4440

tgtaccccgt agacagggcc cagaggttcc cttgtccttg ccctccccca caccacccat     4500

gatcactgtc caacataaag ggttcagtgt gtacgtggtc atgtgttgtc cttacaggat     4560

tcaggtatgt tgccttcacg gttttcccca ccccctcctg ccctttatcc tttaggccgt     4620

gtggccatga acctggaaga agtgatcgtt tgcacttgag tgctacactc ttgcaccttt     4680

ccaaagtaag ctggtttgga ggtcctgttg tcatgtacga gactgtcacc agttaccgcg     4740

ctctgtttga aacatgtctt tgtattccta atgacttcag ttagagtaag gagaatagct     4800

gttaatatgg atgtcaggta cttaaggggc cacaccattg agaattttgt cttggatatt     4860

cttgaaagtt tatattttta taattttttt tacatcagat gtcagatgtt tctttcagtt     4920

gcttgatgtt tggaattatt atgtggcttt ttttgtaaat attgaaatgt agcaataatg     4980

tcttttgaat attcctgagc ccatgagtcc ctgaaaatat tttttatata tacagtaact     5040

ttatgtgtaa ataatacgct gtgcaagttt aaacatgtca cgttacatgt gggttttttc     5100

tgatatgttg tccaactgtt gacagttctg aagaattcta ataaaaatgt aaatatataa     5160

atcaaaaaaa aaaaaaaaaa aaaaaaaaa                                       5189


<210>  2
<211>  975
<212>  PRT
<213>  Mouse

<400>  2

Met Arg Gly Ala Arg Gly Ala Trp Asp Leu Leu Cys Val Leu Leu Val 
1               5                   10                  15      


Leu Leu Arg Gly Gln Thr Ala Thr Ser Gln Pro Ser Ala Ser Pro Gly 
            20                  25                  30          


Glu Pro Ser Pro Pro Ser Ile His Pro Ala Gln Ser Glu Leu Ile Val 
        35                  40                  45              


Glu Ala Gly Asp Thr Leu Ser Leu Thr Cys Ile Asp Pro Asp Phe Val 
    50                  55                  60                  


Arg Trp Thr Phe Lys Thr Tyr Phe Asn Glu Met Val Glu Asn Lys Lys 
65                  70                  75                  80  


Asn Glu Trp Ile Gln Glu Lys Ala Glu Ala Thr Arg Thr Gly Thr Tyr 
                85                  90                  95      


Thr Cys Ser Asn Ser Asn Gly Leu Thr Ser Ser Ile Tyr Val Phe Val 
            100                 105                 110         


Arg Asp Pro Ala Lys Leu Phe Leu Val Gly Leu Pro Leu Phe Gly Lys 
        115                 120                 125             


Glu Asp Ser Asp Ala Leu Val Arg Cys Pro Leu Thr Asp Pro Gln Val 
    130                 135                 140                 


Ser Asn Tyr Ser Leu Ile Glu Cys Asp Gly Lys Ser Leu Pro Thr Asp 
145                 150                 155                 160 


Leu Thr Phe Val Pro Asn Pro Lys Ala Gly Ile Thr Ile Lys Asn Val 
                165                 170                 175     


Lys Arg Ala Tyr His Arg Leu Cys Val Arg Cys Ala Ala Gln Arg Asp 
            180                 185                 190         


Gly Thr Trp Leu His Ser Asp Lys Phe Thr Leu Lys Val Arg Ala Ala 
        195                 200                 205             


Ile Lys Ala Ile Pro Val Val Ser Val Pro Glu Thr Ser His Leu Leu 
    210                 215                 220                 


Lys Lys Gly Asp Thr Phe Thr Val Val Cys Thr Ile Lys Asp Val Ser 
225                 230                 235                 240 


Thr Ser Val Asn Ser Met Trp Leu Lys Met Asn Pro Gln Pro Gln His 
                245                 250                 255     


Ile Ala Gln Val Lys His Asn Ser Trp His Arg Gly Asp Phe Asn Tyr 
            260                 265                 270         


Glu Arg Gln Glu Thr Leu Thr Ile Ser Ser Ala Arg Val Asp Asp Ser 
        275                 280                 285             


Gly Val Phe Met Cys Tyr Ala Asn Asn Thr Phe Gly Ser Ala Asn Val 
    290                 295                 300                 


Thr Thr Thr Leu Lys Val Val Glu Lys Gly Phe Ile Asn Ile Ser Pro 
305                 310                 315                 320 


Val Lys Asn Thr Thr Val Phe Val Thr Asp Gly Glu Asn Val Asp Leu 
                325                 330                 335     


Val Val Glu Tyr Glu Ala Tyr Pro Lys Pro Glu His Gln Gln Trp Ile 
            340                 345                 350         


Tyr Met Asn Arg Thr Ser Ala Asn Lys Gly Lys Asp Tyr Val Lys Ser 
        355                 360                 365             


Asp Asn Lys Ser Asn Ile Arg Tyr Val Asn Gln Leu Arg Leu Thr Arg 
    370                 375                 380                 


Leu Lys Gly Thr Glu Gly Gly Thr Tyr Thr Phe Leu Val Ser Asn Ser 
385                 390                 395                 400 


Asp Ala Ser Ala Ser Val Thr Phe Asn Val Tyr Val Asn Thr Lys Pro 
                405                 410                 415     


Glu Ile Leu Thr Tyr Asp Arg Leu Ile Asn Gly Met Leu Gln Cys Val 
            420                 425                 430         


Ala Glu Gly Phe Pro Glu Pro Thr Ile Asp Trp Tyr Phe Cys Thr Gly 
        435                 440                 445             


Ala Glu Gln Arg Cys Thr Thr Pro Val Ser Pro Val Asp Val Gln Val 
    450                 455                 460                 


Gln Asn Val Ser Val Ser Pro Phe Gly Lys Leu Val Val Gln Ser Ser 
465                 470                 475                 480 


Ile Asp Ser Ser Val Phe Arg His Asn Gly Thr Val Glu Cys Lys Ala 
                485                 490                 495     


Ser Asn Asp Val Gly Lys Ser Ser Ala Phe Phe Asn Phe Ala Phe Lys 
            500                 505                 510         


Glu Gln Ile Gln Ala His Thr Leu Phe Thr Pro Leu Leu Ile Gly Phe 
        515                 520                 525             


Val Val Ala Ala Gly Ala Met Gly Ile Ile Val Met Val Leu Thr Tyr 
    530                 535                 540                 


Lys Tyr Leu Gln Lys Pro Met Tyr Glu Val Gln Trp Lys Val Val Glu 
545                 550                 555                 560 


Glu Ile Asn Gly Asn Asn Tyr Val Tyr Ile Asp Pro Thr Gln Leu Pro 
                565                 570                 575     


Tyr Asp His Lys Trp Glu Phe Pro Arg Asn Arg Leu Ser Phe Gly Lys 
            580                 585                 590         


Thr Leu Gly Ala Gly Ala Phe Gly Lys Val Val Glu Ala Thr Ala Tyr 
        595                 600                 605             


Gly Leu Ile Lys Ser Asp Ala Ala Met Thr Val Ala Val Lys Met Leu 
    610                 615                 620                 


Lys Pro Ser Ala His Leu Thr Glu Arg Glu Ala Leu Met Ser Glu Leu 
625                 630                 635                 640 


Lys Val Leu Ser Tyr Leu Gly Asn His Met Asn Ile Val Asn Leu Leu 
                645                 650                 655     


Gly Ala Cys Thr Val Gly Gly Pro Thr Leu Val Ile Thr Glu Tyr Cys 
            660                 665                 670         


Cys Tyr Gly Asp Leu Leu Asn Phe Leu Arg Arg Lys Arg Asp Ser Phe 
        675                 680                 685             


Ile Phe Ser Lys Gln Glu Glu Gln Ala Glu Ala Ala Leu Tyr Lys Asn 
    690                 695                 700                 


Leu Leu His Ser Thr Glu Pro Ser Cys Asp Ser Ser Asn Glu Tyr Met 
705                 710                 715                 720 


Asp Met Lys Pro Gly Val Ser Tyr Val Val Pro Thr Lys Thr Asp Lys 
                725                 730                 735     


Arg Arg Ser Ala Arg Ile Asp Ser Tyr Ile Glu Arg Asp Val Thr Pro 
            740                 745                 750         


Ala Ile Met Glu Asp Asp Glu Leu Ala Leu Asp Leu Asp Asp Leu Leu 
        755                 760                 765             


Ser Phe Ser Tyr Gln Val Ala Lys Gly Met Ala Phe Leu Ala Ser Lys 
    770                 775                 780                 


Asn Cys Ile His Arg Asp Leu Ala Ala Arg Asn Ile Leu Leu Thr His 
785                 790                 795                 800 


Gly Arg Ile Thr Lys Ile Cys Asp Phe Gly Leu Ala Arg Asp Ile Arg 
                805                 810                 815     


Asn Asp Ser Asn Tyr Val Val Lys Gly Asn Ala Arg Leu Pro Val Lys 
            820                 825                 830         


Trp Met Ala Pro Glu Ser Ile Phe Ser Cys Val Tyr Thr Phe Glu Ser 
        835                 840                 845             


Asp Val Trp Ser Tyr Gly Ile Phe Leu Trp Glu Leu Phe Ser Leu Gly 
    850                 855                 860                 


Ser Ser Pro Tyr Pro Gly Met Pro Val Asp Ser Lys Phe Tyr Lys Met 
865                 870                 875                 880 


Ile Lys Glu Gly Phe Arg Met Val Ser Pro Glu His Ala Pro Ala Glu 
                885                 890                 895     


Met Tyr Asp Val Met Lys Thr Cys Trp Asp Ala Asp Pro Leu Lys Arg 
            900                 905                 910         


Pro Thr Phe Lys Gln Val Val Gln Leu Ile Glu Lys Gln Ile Ser Asp 
        915                 920                 925             


Ser Thr Lys His Ile Tyr Ser Asn Leu Ala Asn Cys Asn Pro Asn Pro 
    930                 935                 940                 


Glu Asn Pro Val Val Val Asp His Ser Val Arg Val Asn Ser Val Gly 
945                 950                 955                 960 


Ser Ser Ala Ser Ser Thr Gln Pro Leu Leu Val His Glu Asp Ala 
                965                 970                 975 


<210>  3
<211>  188
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  partial DNA sequence after the mutation

<400>  3
gctttgtttt gctaaaatac atcactttcc cttttagcaa gtgctcattt aacagaaaga       60

gaagccctaa tgtcggaact gaaggtcctg agctacctgg gcaatcacat gaatattgtg      120

aatctgcttg gcgcatgcat ggttggaggt gagtctcgag ttgccatctc tctggttatc      180

aaagtatg                                                               188


<210>  4
<211>  975
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  amino acid sequence after the mutation

<400>  4

Met Arg Gly Ala Arg Gly Ala Trp Asp Leu Leu Cys Val Leu Leu Val 
1               5                   10                  15      


Leu Leu Arg Gly Gln Thr Ala Thr Ser Gln Pro Ser Ala Ser Pro Gly 
            20                  25                  30          


Glu Pro Ser Pro Pro Ser Ile His Pro Ala Gln Ser Glu Leu Ile Val 
        35                  40                  45              


Glu Ala Gly Asp Thr Leu Ser Leu Thr Cys Ile Asp Pro Asp Phe Val 
    50                  55                  60                  


Arg Trp Thr Phe Lys Thr Tyr Phe Asn Glu Met Val Glu Asn Lys Lys 
65                  70                  75                  80  


Asn Glu Trp Ile Gln Glu Lys Ala Glu Ala Thr Arg Thr Gly Thr Tyr 
                85                  90                  95      


Thr Cys Ser Asn Ser Asn Gly Leu Thr Ser Ser Ile Tyr Val Phe Val 
            100                 105                 110         


Arg Asp Pro Ala Lys Leu Phe Leu Val Gly Leu Pro Leu Phe Gly Lys 
        115                 120                 125             


Glu Asp Ser Asp Ala Leu Val Arg Cys Pro Leu Thr Asp Pro Gln Val 
    130                 135                 140                 


Ser Asn Tyr Ser Leu Ile Glu Cys Asp Gly Lys Ser Leu Pro Thr Asp 
145                 150                 155                 160 


Leu Thr Phe Val Pro Asn Pro Lys Ala Gly Ile Thr Ile Lys Asn Val 
                165                 170                 175     


Lys Arg Ala Tyr His Arg Leu Cys Val Arg Cys Ala Ala Gln Arg Asp 
            180                 185                 190         


Gly Thr Trp Leu His Ser Asp Lys Phe Thr Leu Lys Val Arg Ala Ala 
        195                 200                 205             


Ile Lys Ala Ile Pro Val Val Ser Val Pro Glu Thr Ser His Leu Leu 
    210                 215                 220                 


Lys Lys Gly Asp Thr Phe Thr Val Val Cys Thr Ile Lys Asp Val Ser 
225                 230                 235                 240 


Thr Ser Val Asn Ser Met Trp Leu Lys Met Asn Pro Gln Pro Gln His 
                245                 250                 255     


Ile Ala Gln Val Lys His Asn Ser Trp His Arg Gly Asp Phe Asn Tyr 
            260                 265                 270         


Glu Arg Gln Glu Thr Leu Thr Ile Ser Ser Ala Arg Val Asp Asp Ser 
        275                 280                 285             


Gly Val Phe Met Cys Tyr Ala Asn Asn Thr Phe Gly Ser Ala Asn Val 
    290                 295                 300                 


Thr Thr Thr Leu Lys Val Val Glu Lys Gly Phe Ile Asn Ile Ser Pro 
305                 310                 315                 320 


Val Lys Asn Thr Thr Val Phe Val Thr Asp Gly Glu Asn Val Asp Leu 
                325                 330                 335     


Val Val Glu Tyr Glu Ala Tyr Pro Lys Pro Glu His Gln Gln Trp Ile 
            340                 345                 350         


Tyr Met Asn Arg Thr Ser Ala Asn Lys Gly Lys Asp Tyr Val Lys Ser 
        355                 360                 365             


Asp Asn Lys Ser Asn Ile Arg Tyr Val Asn Gln Leu Arg Leu Thr Arg 
    370                 375                 380                 


Leu Lys Gly Thr Glu Gly Gly Thr Tyr Thr Phe Leu Val Ser Asn Ser 
385                 390                 395                 400 


Asp Ala Ser Ala Ser Val Thr Phe Asn Val Tyr Val Asn Thr Lys Pro 
                405                 410                 415     


Glu Ile Leu Thr Tyr Asp Arg Leu Ile Asn Gly Met Leu Gln Cys Val 
            420                 425                 430         


Ala Glu Gly Phe Pro Glu Pro Thr Ile Asp Trp Tyr Phe Cys Thr Gly 
        435                 440                 445             


Ala Glu Gln Arg Cys Thr Thr Pro Val Ser Pro Val Asp Val Gln Val 
    450                 455                 460                 


Gln Asn Val Ser Val Ser Pro Phe Gly Lys Leu Val Val Gln Ser Ser 
465                 470                 475                 480 


Ile Asp Ser Ser Val Phe Arg His Asn Gly Thr Val Glu Cys Lys Ala 
                485                 490                 495     


Ser Asn Asp Val Gly Lys Ser Ser Ala Phe Phe Asn Phe Ala Phe Lys 
            500                 505                 510         


Glu Gln Ile Gln Ala His Thr Leu Phe Thr Pro Leu Leu Ile Gly Phe 
        515                 520                 525             


Val Val Ala Ala Gly Ala Met Gly Ile Ile Val Met Val Leu Thr Tyr 
    530                 535                 540                 


Lys Tyr Leu Gln Lys Pro Met Tyr Glu Val Gln Trp Lys Val Val Glu 
545                 550                 555                 560 


Glu Ile Asn Gly Asn Asn Tyr Val Tyr Ile Asp Pro Thr Gln Leu Pro 
                565                 570                 575     


Tyr Asp His Lys Trp Glu Phe Pro Arg Asn Arg Leu Ser Phe Gly Lys 
            580                 585                 590         


Thr Leu Gly Ala Gly Ala Phe Gly Lys Val Val Glu Ala Thr Ala Tyr 
        595                 600                 605             


Gly Leu Ile Lys Ser Asp Ala Ala Met Thr Val Ala Val Lys Met Leu 
    610                 615                 620                 


Lys Pro Ser Ala His Leu Thr Glu Arg Glu Ala Leu Met Ser Glu Leu 
625                 630                 635                 640 


Lys Val Leu Ser Tyr Leu Gly Asn His Met Asn Ile Val Asn Leu Leu 
                645                 650                 655     


Gly Ala Cys Met Val Gly Gly Pro Thr Leu Val Ile Thr Glu Tyr Cys 
            660                 665                 670         


Cys Tyr Gly Asp Leu Leu Asn Phe Leu Arg Arg Lys Arg Asp Ser Phe 
        675                 680                 685             


Ile Phe Ser Lys Gln Glu Glu Gln Ala Glu Ala Ala Leu Tyr Lys Asn 
    690                 695                 700                 


Leu Leu His Ser Thr Glu Pro Ser Cys Asp Ser Ser Asn Glu Tyr Met 
705                 710                 715                 720 


Asp Met Lys Pro Gly Val Ser Tyr Val Val Pro Thr Lys Thr Asp Lys 
                725                 730                 735     


Arg Arg Ser Ala Arg Ile Asp Ser Tyr Ile Glu Arg Asp Val Thr Pro 
            740                 745                 750         


Ala Ile Met Glu Asp Asp Glu Leu Ala Leu Asp Leu Asp Asp Leu Leu 
        755                 760                 765             


Ser Phe Ser Tyr Gln Val Ala Lys Gly Met Ala Phe Leu Ala Ser Lys 
    770                 775                 780                 


Asn Cys Ile His Arg Asp Leu Ala Ala Arg Asn Ile Leu Leu Thr His 
785                 790                 795                 800 


Gly Arg Ile Thr Lys Ile Cys Asp Phe Gly Leu Ala Arg Asp Ile Arg 
                805                 810                 815     


Asn Asp Ser Asn Tyr Val Val Lys Gly Asn Ala Arg Leu Pro Val Lys 
            820                 825                 830         


Trp Met Ala Pro Glu Ser Ile Phe Ser Cys Val Tyr Thr Phe Glu Ser 
        835                 840                 845             


Asp Val Trp Ser Tyr Gly Ile Phe Leu Trp Glu Leu Phe Ser Leu Gly 
    850                 855                 860                 


Ser Ser Pro Tyr Pro Gly Met Pro Val Asp Ser Lys Phe Tyr Lys Met 
865                 870                 875                 880 


Ile Lys Glu Gly Phe Arg Met Val Ser Pro Glu His Ala Pro Ala Glu 
                885                 890                 895     


Met Tyr Asp Val Met Lys Thr Cys Trp Asp Ala Asp Pro Leu Lys Arg 
            900                 905                 910         


Pro Thr Phe Lys Gln Val Val Gln Leu Ile Glu Lys Gln Ile Ser Asp 
        915                 920                 925             


Ser Thr Lys His Ile Tyr Ser Asn Leu Ala Asn Cys Asn Pro Asn Pro 
    930                 935                 940                 


Glu Asn Pro Val Val Val Asp His Ser Val Arg Val Asn Ser Val Gly 
945                 950                 955                 960 


Ser Ser Ala Ser Ser Thr Gln Pro Leu Leu Val His Glu Asp Ala 
                965                 970                 975 


<210>  5
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA

<400>  5
ccaccgtgca tgcgccaagc agg                                               23


<210>  6
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA

<400>  6
cctgcttggc gcatgcacgg tgg                                               23


<210>  7
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA

<400>  7
ctgcttggcg catgcacggt ggg                                               23


<210>  8
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA

<400>  8
gaacctgctt ggcgcatgca cgg                                               23


<210>  9
<211>  132
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA fragment containing the T7 promoter and sgRNA scaffold

<400>  9
gaattctaat acgactcact atagggggtc ttcgagaaga cctgttttag agctagaaat       60

agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct      120

tttaaaggat cc                                                          132


<210>  10
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Upstream

<400>  10
ggcctgcttg gcgcatgcac gg                                                22


<210>  11
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward oligonucleotide

<400>  11
caccggcctg cttggcgcat gcacgg                                            26


<210>  12
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Downstream

<400>  12
ccgtgcatgc gccaagcagg cc                                                22


<210>  13
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse oligonucleotide

<400>  13
aaacccgtgc atgcgccaag caggcc                                            26


<210>  14
<211>  952
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  5 homologous arm

<400>  14
tggttccctt ccttgcagag caaatccagg cccacactct gttcacgccg ctgctcattg       60

gctttgtggt tgcagctggc gcgatgggga tcattgtgat ggtgctcacc tacaaatatt      120

tgcaggtgag cattgaattg ttctcttcct ggggacgcca aggcggcagg gcaggcactg      180

attgttcagc gggtgacaca tctttctttt cctttctcct ccagaaaccc atgtatgaag      240

tacaatggaa ggttgtcgag gagataaatg gaaacaatta tgtttacata gacccgacgc      300

aacttcctta tgatcacaaa tgggagtttc ccagaaacag gctgagtttt ggtcagtgtg      360

aggcaggggc tttccacgaa gaagcccttt tgtgtacgcg taaccgtgat tttttttttt      420

taaggaaccc gcagtggctt cctttgtctt gtttcaccta aaacaagaag tgctttctgt      480

caaactgcac aggagttggc agggttagca gaaagagccc cgtagagaag tggctccaga      540

cacacatgtc ttccattgcc ttgttgattt gggaacctca caaactgttg gttggtcttc      600

ccactgacat gaccctgcct tcttccttcc taaaggaaag acattgggag ctggtgcctt      660

cgggaaggtc gttgaggcca ctgcatatgg cttgattaag tcggatgctg ccatgacagt      720

tgccgtgaag atgctcaaac gtaagtgttt cgggggcact caccccctgt acccaccacc      780

agtttatcta ataggttctt cccccctgcc cccacgcttt gttttgctaa aatacatcac      840

tttccctttt agcaagtgct catttaacag aaagagaagc cctaatgtcg gaactgaagg      900

tcctgagcta cctgggcaat cacatgaata ttgtgaatct gcttggcgca tg              952


<210>  15
<211>  947
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3 homologous arm

<400>  15
ggttggaggt gagtctcgag ttgccatctc tctggttatc aaagtatgat ttaggcagtg       60

cctagctata agctacttga gagatttttt ttttttaacc aagcattttg ttctattttg      120

ctaggtttcg ttttccacag tgttcctcag aatgtagatt tttataattt ctatgctggg      180

agccttcaga ctcatgtctt gtcatgattt tgcaatgtag gttgtagttg ttccgctgtg      240

tggtcgagtg ttggaactac tcatcacagt tgtgtttggt acccattact cagattctta      300

tttctcccct ccctctccac agtcctaagg acctctattc tgccctcagt ggatcaacat      360

tttaaactcc cacaagcttt tgggaacaca caaggtttct ctttatgcag agccacttac      420

accccgcctt tcctcattag gcattatatg tgtgtattga attataatac tctccccctt      480

aaatggcata agcagtgggg ggcaggtagc tttatataat gcggggggcc gtaatcagtc      540

atttgagagt taatgttgat tttccttcta taactgagag tggccccagc ctcccagttc      600

tcaactgtac accaagtaga tccagctttt cactcccaca aaagcaagct ggcggctcta      660

gcccctaagt cagagtgcca ccaagggtgc gcaggcacac tgttctacag aacaaagctg      720

tgtagggtga gctggtgctg tctgtcctga gagagccatg ggatggtagg cagcttagac      780

aacgtctccg gacagcccac agacccggca ctctgcacac catgaattgt tcatgcgaca      840

gtatctaccc atggcaatta gttgcctgtt aagttcggaa ctgcgctttt aaacatcctg      900

aggaaccact cgtgttttca agtagtcatg ctcagaaaaa cgggccg                    947


<210>  16
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer 

<400>  16
actgttggtt ggtcttccca ctgac                                             25


<210>  17
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer 

<400>  17
agcctagtag ggaagtaacc aggga                                             25


<210>  18
<211>  979
<212>  PRT
<213>  Mouse

<400>  18

Met Arg Gly Ala Arg Gly Ala Trp Asp Leu Leu Cys Val Leu Leu Val 
1               5                   10                  15      


Leu Leu Arg Gly Gln Thr Ala Thr Ser Gln Pro Ser Ala Ser Pro Gly 
            20                  25                  30          


Glu Pro Ser Pro Pro Ser Ile His Pro Ala Gln Ser Glu Leu Ile Val 
        35                  40                  45              


Glu Ala Gly Asp Thr Leu Ser Leu Thr Cys Ile Asp Pro Asp Phe Val 
    50                  55                  60                  


Arg Trp Thr Phe Lys Thr Tyr Phe Asn Glu Met Val Glu Asn Lys Lys 
65                  70                  75                  80  


Asn Glu Trp Ile Gln Glu Lys Ala Glu Ala Thr Arg Thr Gly Thr Tyr 
                85                  90                  95      


Thr Cys Ser Asn Ser Asn Gly Leu Thr Ser Ser Ile Tyr Val Phe Val 
            100                 105                 110         


Arg Asp Pro Ala Lys Leu Phe Leu Val Gly Leu Pro Leu Phe Gly Lys 
        115                 120                 125             


Glu Asp Ser Asp Ala Leu Val Arg Cys Pro Leu Thr Asp Pro Gln Val 
    130                 135                 140                 


Ser Asn Tyr Ser Leu Ile Glu Cys Asp Gly Lys Ser Leu Pro Thr Asp 
145                 150                 155                 160 


Leu Thr Phe Val Pro Asn Pro Lys Ala Gly Ile Thr Ile Lys Asn Val 
                165                 170                 175     


Lys Arg Ala Tyr His Arg Leu Cys Val Arg Cys Ala Ala Gln Arg Asp 
            180                 185                 190         


Gly Thr Trp Leu His Ser Asp Lys Phe Thr Leu Lys Val Arg Ala Ala 
        195                 200                 205             


Ile Lys Ala Ile Pro Val Val Ser Val Pro Glu Thr Ser His Leu Leu 
    210                 215                 220                 


Lys Lys Gly Asp Thr Phe Thr Val Val Cys Thr Ile Lys Asp Val Ser 
225                 230                 235                 240 


Thr Ser Val Asn Ser Met Trp Leu Lys Met Asn Pro Gln Pro Gln His 
                245                 250                 255     


Ile Ala Gln Val Lys His Asn Ser Trp His Arg Gly Asp Phe Asn Tyr 
            260                 265                 270         


Glu Arg Gln Glu Thr Leu Thr Ile Ser Ser Ala Arg Val Asp Asp Ser 
        275                 280                 285             


Gly Val Phe Met Cys Tyr Ala Asn Asn Thr Phe Gly Ser Ala Asn Val 
    290                 295                 300                 


Thr Thr Thr Leu Lys Val Val Glu Lys Gly Phe Ile Asn Ile Ser Pro 
305                 310                 315                 320 


Val Lys Asn Thr Thr Val Phe Val Thr Asp Gly Glu Asn Val Asp Leu 
                325                 330                 335     


Val Val Glu Tyr Glu Ala Tyr Pro Lys Pro Glu His Gln Gln Trp Ile 
            340                 345                 350         


Tyr Met Asn Arg Thr Ser Ala Asn Lys Gly Lys Asp Tyr Val Lys Ser 
        355                 360                 365             


Asp Asn Lys Ser Asn Ile Arg Tyr Val Asn Gln Leu Arg Leu Thr Arg 
    370                 375                 380                 


Leu Lys Gly Thr Glu Gly Gly Thr Tyr Thr Phe Leu Val Ser Asn Ser 
385                 390                 395                 400 


Asp Ala Ser Ala Ser Val Thr Phe Asn Val Tyr Val Asn Thr Lys Pro 
                405                 410                 415     


Glu Ile Leu Thr Tyr Asp Arg Leu Ile Asn Gly Met Leu Gln Cys Val 
            420                 425                 430         


Ala Glu Gly Phe Pro Glu Pro Thr Ile Asp Trp Tyr Phe Cys Thr Gly 
        435                 440                 445             


Ala Glu Gln Arg Cys Thr Thr Pro Val Ser Pro Val Asp Val Gln Val 
    450                 455                 460                 


Gln Asn Val Ser Val Ser Pro Phe Gly Lys Leu Val Val Gln Ser Ser 
465                 470                 475                 480 


Ile Asp Ser Ser Val Phe Arg His Asn Gly Thr Val Glu Cys Lys Ala 
                485                 490                 495     


Ser Asn Asp Val Gly Lys Ser Ser Ala Phe Phe Asn Phe Ala Phe Lys 
            500                 505                 510         


Gly Asn Asn Lys Glu Gln Ile Gln Ala His Thr Leu Phe Thr Pro Leu 
        515                 520                 525             


Leu Ile Gly Phe Val Val Ala Ala Gly Ala Met Gly Ile Ile Val Met 
    530                 535                 540                 


Val Leu Thr Tyr Lys Tyr Leu Gln Lys Pro Met Tyr Glu Val Gln Trp 
545                 550                 555                 560 


Lys Val Val Glu Glu Ile Asn Gly Asn Asn Tyr Val Tyr Ile Asp Pro 
                565                 570                 575     


Thr Gln Leu Pro Tyr Asp His Lys Trp Glu Phe Pro Arg Asn Arg Leu 
            580                 585                 590         


Ser Phe Gly Lys Thr Leu Gly Ala Gly Ala Phe Gly Lys Val Val Glu 
        595                 600                 605             


Ala Thr Ala Tyr Gly Leu Ile Lys Ser Asp Ala Ala Met Thr Val Ala 
    610                 615                 620                 


Val Lys Met Leu Lys Pro Ser Ala His Leu Thr Glu Arg Glu Ala Leu 
625                 630                 635                 640 


Met Ser Glu Leu Lys Val Leu Ser Tyr Leu Gly Asn His Met Asn Ile 
                645                 650                 655     


Val Asn Leu Leu Gly Ala Cys Thr Val Gly Gly Pro Thr Leu Val Ile 
            660                 665                 670         


Thr Glu Tyr Cys Cys Tyr Gly Asp Leu Leu Asn Phe Leu Arg Arg Lys 
        675                 680                 685             


Arg Asp Ser Phe Ile Phe Ser Lys Gln Glu Glu Gln Ala Glu Ala Ala 
    690                 695                 700                 


Leu Tyr Lys Asn Leu Leu His Ser Thr Glu Pro Ser Cys Asp Ser Ser 
705                 710                 715                 720 


Asn Glu Tyr Met Asp Met Lys Pro Gly Val Ser Tyr Val Val Pro Thr 
                725                 730                 735     


Lys Thr Asp Lys Arg Arg Ser Ala Arg Ile Asp Ser Tyr Ile Glu Arg 
            740                 745                 750         


Asp Val Thr Pro Ala Ile Met Glu Asp Asp Glu Leu Ala Leu Asp Leu 
        755                 760                 765             


Asp Asp Leu Leu Ser Phe Ser Tyr Gln Val Ala Lys Gly Met Ala Phe 
    770                 775                 780                 


Leu Ala Ser Lys Asn Cys Ile His Arg Asp Leu Ala Ala Arg Asn Ile 
785                 790                 795                 800 


Leu Leu Thr His Gly Arg Ile Thr Lys Ile Cys Asp Phe Gly Leu Ala 
                805                 810                 815     


Arg Asp Ile Arg Asn Asp Ser Asn Tyr Val Val Lys Gly Asn Ala Arg 
            820                 825                 830         


Leu Pro Val Lys Trp Met Ala Pro Glu Ser Ile Phe Ser Cys Val Tyr 
        835                 840                 845             


Thr Phe Glu Ser Asp Val Trp Ser Tyr Gly Ile Phe Leu Trp Glu Leu 
    850                 855                 860                 


Phe Ser Leu Gly Ser Ser Pro Tyr Pro Gly Met Pro Val Asp Ser Lys 
865                 870                 875                 880 


Phe Tyr Lys Met Ile Lys Glu Gly Phe Arg Met Val Ser Pro Glu His 
                885                 890                 895     


Ala Pro Ala Glu Met Tyr Asp Val Met Lys Thr Cys Trp Asp Ala Asp 
            900                 905                 910         


Pro Leu Lys Arg Pro Thr Phe Lys Gln Val Val Gln Leu Ile Glu Lys 
        915                 920                 925             


Gln Ile Ser Asp Ser Thr Lys His Ile Tyr Ser Asn Leu Ala Asn Cys 
    930                 935                 940                 


Asn Pro Asn Pro Glu Asn Pro Val Val Val Asp His Ser Val Arg Val 
945                 950                 955                 960 


Asn Ser Val Gly Ser Ser Ala Ser Ser Thr Gln Pro Leu Leu Val His 
                965                 970                 975     


Glu Asp Ala 
            

