               SEQUENCE LISTING

<110> Max-Planck-Gesellschaft zur Foerderung der Wissenschaften e.V.

<120> Targeted In Situ Protein Diversification by Site Directed DNA cleavage and Repair

<130> Z1423 PCT S3

<150> EP 16 18 1872.9
<151> 2016-07-29

<160> 95

<170> BiSSAP 1.3

<210> 1
<211> 18
<212> PRT
<213> Artificial Sequence

<220> 
<223> 2A peptide T2A

<400> 1
Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro 
1               5                   10                  15      
Gly Pro 
        

<210> 2
<211> 19
<212> PRT
<213> Artificial Sequence

<220> 
<223> 2A peptide P2A

<400> 2
Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn 
1               5                   10                  15      
Pro Gly Pro 
            

<210> 3
<211> 20
<212> PRT
<213> Artificial Sequence

<220> 
<223> 2A peptide E2A

<400> 3
Gln Cys Thr Asn Tyr Ala Leu Leu Lys Leu Ala Gly Asp Val Glu Ser 
1               5                   10                  15      
Asn Pro Gly Pro 
            20  

<210> 4
<211> 22
<212> PRT
<213> Artificial Sequence

<220> 
<223> 2A peptide F2A

<400> 4
Val Lys Gln Thr Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp Val 
1               5                   10                  15      
Glu Ser Asn Pro Gly Pro 
            20          

<210> 5
<211> 7
<212> PRT
<213> Artificial Sequence

<220> 
<223> Target site of TEV protease

<220> 
<221> UNSURE
<222> 2
<223> note: Xaa = any amino acid

<220> 
<221> UNSURE
<222> 3
<223> note: Xaa = any amino acid

<220> 
<221> UNSURE
<222> 5
<223> note: Xaa = any amino acid

<220> 
<221> VARIANT
<222> 7
<223> Replace: Ser

<400> 5
Glu Xaa Xaa Tyr Xaa Gln Gly 
1               5           

<210> 6
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> Target Site of Genenase I

<400> 6
Pro Gly Ala Ala His Tyr 
1               5       

<210> 7
<211> 5
<212> PRT
<213> Artificial Sequence

<220> 
<223> Target site of Enterokinase

<400> 7
Asp Asp Asp Asp Lys 
1               5   

<210> 8
<211> 8
<212> PRT
<213> Artificial Sequence

<220> 
<223> Target site of Human Rhinovirus (HRV) 3C Protease

<400> 8
Leu Glu Val Leu Phe Gln Gly Pro 
1               5               

<210> 9
<211> 4
<212> PRT
<213> Artificial Sequence

<220> 
<223> Target site of Factor Xa

<220> 
<221> VARIANT
<222> 2
<223> Replace: Asp

<400> 9
Ile Glu Gly Arg 
1               

<210> 10
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> Target site of Thrombin

<400> 10
Leu Val Pro Arg Gly Ser 
1               5       

<210> 11
<211> 12
<212> DNA
<213> Artificial Sequence

<220> 
<223> Preferred direct repeat (DR) sequence for use with SpCas9 or
      SaCas9 nuclease

<400> 11
gttttagagc ta                                                        12


<210> 12
<211> 31
<212> DNA
<213> Artificial Sequence

<220> 
<223> Preferred tracrRNA sequence for use with the SpCas9 or SaCas9
      nuclease

<400> 12
tagcaagtta aaataaggct agtccgtttt t                                   31


<210> 13
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> Forward primer for site-directed mutagenesis

<400> 13
tcgctgaccg ctgcggacgc aggtcgaaga agacttacc                            39


<210> 14
<211> 25
<212> DNA
<213> Artificial Sequence

<220> 
<223> Reverse primer for site-directed mutagenesis

<400> 14
gtccgcagcg gtcagcgagt tggtc                                          25


<210> 15
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> Forward amplification primer

<400> 15
tcgctgaccg ctgcggacgc aggtcgaaga agacttacc                            39


<210> 16
<211> 49
<212> DNA
<213> Artificial Sequence

<220> 
<223> Reverse amplification primer

<400> 16
cggccgccac tgtgctggat ctattatcac ttgtacagct cgtccatgc                 49


<210> 17
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Pre-annealed forward primer

<400> 17
caccgcgctg accgctgcgg acgc                                           24


<210> 18
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Pre-annealed reverse primer

<400> 18
aaacgcgtcc gcagcggtca gcgc                                           24


<210> 19
<211> 198
<212> PRT
<213> Artificial Sequence

<220> 
<223> FokI nuclease

<400> 19
Gly Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu 
1               5                   10                  15      
Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu 
            20                  25                  30          
Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met 
        35                  40                  45              
Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly 
    50                  55                  60                  
Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp 
65                  70                  75                  80  
Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu 
                85                  90                  95      
Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln 
            100                 105                 110         
Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro 
        115                 120                 125             
Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys 
    130                 135                 140                 
Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys 
145                 150                 155                 160 
Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met 
                165                 170                 175     
Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn 
            180                 185                 190         
Asn Gly Glu Ile Asn Phe 
        195             

<210> 20
<211> 1022
<212> PRT
<213> Artificial Sequence

<220> 
<223> megaTAL endonuclease

<400> 20
Val Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile 
1               5                   10                  15      
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val 
            20                  25                  30          
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro 
        35                  40                  45              
Ala Ala Leu Gly Thr Val Ala Val Thr Tyr Gln His Ile Ile Thr Ala 
    50                  55                  60                  
Leu Pro Glu Ala Thr His Glu Asp Ile Val Gly Val Gly Lys Gln Trp 
65                  70                  75                  80  
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Asp Ala Gly Glu Leu 
                85                  90                  95      
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Val Lys Ile Ala 
            100                 105                 110         
Lys Arg Gly Gly Val Thr Ala Met Glu Ala Val His Ala Ser Arg Asn 
        115                 120                 125             
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala 
    130                 135                 140                 
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg 
145                 150                 155                 160 
Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val 
                165                 170                 175     
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val 
            180                 185                 190         
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp 
        195                 200                 205             
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu 
    210                 215                 220                 
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr 
225                 230                 235                 240 
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala 
                245                 250                 255     
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly 
            260                 265                 270         
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys 
        275                 280                 285             
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp 
    290                 295                 300                 
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly 
305                 310                 315                 320 
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys 
                325                 330                 335     
Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His 
            340                 345                 350         
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val 
        355                 360                 365             
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala 
    370                 375                 380                 
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu 
385                 390                 395                 400 
Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala 
                405                 410                 415     
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg 
            420                 425                 430         
Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val 
        435                 440                 445             
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val 
    450                 455                 460                 
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp 
465                 470                 475                 480 
Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu 
                485                 490                 495     
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr 
            500                 505                 510         
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala 
        515                 520                 525             
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly 
    530                 535                 540                 
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys 
545                 550                 555                 560 
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp 
                565                 570                 575     
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly 
            580                 585                 590         
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys 
        595                 600                 605             
Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn 
    610                 615                 620                 
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val 
625                 630                 635                 640 
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala 
                645                 650                 655     
Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu 
            660                 665                 670         
Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala 
        675                 680                 685             
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala 
    690                 695                 700                 
Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His 
705                 710                 715                 720 
Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Met Asp Ala Val 
                725                 730                 735     
Lys Lys Gly Leu Pro His Ala Pro Glu Leu Ile Arg Arg Val Asn Arg 
            740                 745                 750         
Arg Ile Gly Glu Arg Thr Ser His Arg Val Ala Ile Ser Arg Val Gly 
        755                 760                 765             
Gly Ser Asp Leu Thr Tyr Ala Tyr Leu Val Gly Leu Tyr Glu Gly Asp 
    770                 775                 780                 
Gly Tyr Phe Ser Ile Thr Lys Lys Gly Lys Tyr Leu Thr Tyr Glu Leu 
785                 790                 795                 800 
Gly Ile Glu Leu Ser Ile Lys Asp Val Gln Leu Ile Tyr Lys Ile Lys 
                805                 810                 815     
Lys Ile Leu Gly Ile Gly Ile Val Ser Phe Arg Lys Arg Asn Glu Ile 
            820                 825                 830         
Glu Met Val Ala Leu Arg Ile Arg Asp Lys Asn His Leu Lys Ser Lys 
        835                 840                 845             
Ile Leu Pro Ile Phe Glu Lys Tyr Pro Met Phe Ser Asn Lys Gln Tyr 
    850                 855                 860                 
Asp Tyr Leu Arg Phe Arg Asn Ala Leu Leu Ser Gly Ile Ile Tyr Leu 
865                 870                 875                 880 
Glu Asp Leu Pro Asp Tyr Thr Arg Ser Asp Glu Pro Leu Asn Ser Ile 
                885                 890                 895     
Glu Ser Ile Ile Asn Thr Ser Tyr Phe Ser Ala Trp Leu Val Gly Phe 
            900                 905                 910         
Ile Glu Ala Glu Gly Cys Phe Ser Val Tyr Lys Leu Asn Lys Asp Asp 
        915                 920                 925             
Asp Tyr Leu Ile Ala Ser Phe Asp Ile Ala Gln Arg Asp Gly Asp Ile 
    930                 935                 940                 
Leu Ile Ser Ala Ile Arg Lys Tyr Leu Ser Phe Thr Thr Lys Val Tyr 
945                 950                 955                 960 
Leu Asp Lys Thr Asn Cys Ser Lys Leu Lys Val Thr Ser Val Arg Ser 
                965                 970                 975     
Val Glu Asn Ile Ile Lys Phe Leu Gln Asn Ala Pro Val Lys Leu Leu 
            980                 985                 990         
Gly Asn Lys Lys Leu Gln Tyr Lys Leu Trp Leu Lys Gln Leu Arg Lys 
        995                 1000                1005            
Ile Ser Arg Tyr Ser Glu Lys Ile Lys Ile Pro Ser Asn Tyr 
    1010                1015                1020        

<210> 21
<211> 1307
<212> PRT
<213> Artificial Sequence

<220> 
<223> AsCpf1

<400> 21
Met Thr Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr 
1               5                   10                  15      
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln 
            20                  25                  30          
Glu Gln Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys 
        35                  40                  45              
Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln 
    50                  55                  60                  
Cys Leu Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile 
65                  70                  75                  80  
Asp Ser Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile 
                85                  90                  95      
Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly 
            100                 105                 110         
Arg Thr Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile 
        115                 120                 125             
Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys 
    130                 135                 140                 
Gln Leu Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg 
145                 150                 155                 160 
Ser Phe Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg 
                165                 170                 175     
Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg 
            180                 185                 190         
Ile Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe 
        195                 200                 205             
Thr Arg Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn 
    210                 215                 220                 
Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val 
225                 230                 235                 240 
Phe Ser Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp 
                245                 250                 255     
Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu 
            260                 265                 270         
Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn 
        275                 280                 285             
Asp Glu Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro 
    290                 295                 300                 
Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu 
305                 310                 315                 320 
Glu Glu Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr 
                325                 330                 335     
Lys Thr Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu 
            340                 345                 350         
Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His 
        355                 360                 365             
Lys Lys Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr 
    370                 375                 380                 
Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys 
385                 390                 395                 400 
Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu 
                405                 410                 415     
Asp Ile Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser 
            420                 425                 430         
Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala 
        435                 440                 445             
Ala Leu Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys 
    450                 455                 460                 
Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu 
465                 470                 475                 480 
Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe 
                485                 490                 495     
Ser Ala Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser 
            500                 505                 510         
Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val 
        515                 520                 525             
Glu Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp 
    530                 535                 540                 
Asp Val Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn 
545                 550                 555                 560 
Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys 
                565                 570                 575     
Ala Leu Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys 
            580                 585                 590         
Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys 
        595                 600                 605             
Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr 
    610                 615                 620                 
Pro Ile Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys 
625                 630                 635                 640 
Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln 
                645                 650                 655     
Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala 
            660                 665                 670         
Leu Cys Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr 
        675                 680                 685             
Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr 
    690                 695                 700                 
Lys Asp Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His 
705                 710                 715                 720 
Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu 
                725                 730                 735     
Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys 
            740                 745                 750         
Gly His His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu 
        755                 760                 765             
Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln 
    770                 775                 780                 
Ala Glu Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His 
785                 790                 795                 800 
Arg Leu Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr 
                805                 810                 815     
Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His 
            820                 825                 830         
Arg Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn 
        835                 840                 845             
Val Ile Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe 
    850                 855                 860                 
Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln 
865                 870                 875                 880 
Ala Ala Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu 
                885                 890                 895     
Lys Glu His Pro Glu Thr Pro Ile Ile Gly Ile Asp Arg Gly Glu Arg 
            900                 905                 910         
Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu 
        915                 920                 925             
Gln Arg Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu 
    930                 935                 940                 
Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val 
945                 950                 955                 960 
Val Gly Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile 
                965                 970                 975     
His Glu Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu 
            980                 985                 990         
Glu Asn Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile Ala Glu 
        995                 1000                1005            
Lys Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu Asn 
    1010                1015                1020                
Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly Val Leu 
1025                1030                1035                1040
Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala Lys Met Gly 
                1045                1050                1055    
Thr Gln Ser Gly Phe Leu Phe Tyr Val Pro Ala Pro Tyr Thr Ser Lys 
            1060                1065                1070        
Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe Val Trp Lys Thr Ile 
        1075                1080                1085            
Lys Asn His Glu Ser Arg Lys His Phe Leu Glu Gly Phe Asp Phe Leu 
    1090                1095                1100                
His Tyr Asp Val Lys Thr Gly Asp Phe Ile Leu His Phe Lys Met Asn 
1105                1110                1115                1120
Arg Asn Leu Ser Phe Gln Arg Gly Leu Pro Gly Phe Met Pro Ala Trp 
                1125                1130                1135    
Asp Ile Val Phe Glu Lys Asn Glu Thr Gln Phe Asp Ala Lys Gly Thr 
            1140                1145                1150        
Pro Phe Ile Ala Gly Lys Arg Ile Val Pro Val Ile Glu Asn His Arg 
        1155                1160                1165            
Phe Thr Gly Arg Tyr Arg Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala 
    1170                1175                1180                
Leu Leu Glu Glu Lys Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu 
1185                1190                1195                1200
Pro Lys Leu Leu Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val 
                1205                1210                1215    
Ala Leu Ile Arg Ser Val Leu Gln Met Arg Asn Ser Asn Ala Ala Thr 
            1220                1225                1230        
Gly Glu Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys 
        1235                1240                1245            
Phe Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp Ala 
    1250                1255                1260                
Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu Asn His 
1265                1270                1275                1280
Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile Ser Asn Gln 
                1285                1290                1295    
Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn 
            1300                1305        

<210> 22
<211> 1228
<212> PRT
<213> Artificial Sequence

<220> 
<223> LbCpf1

<400> 22
Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr 
1               5                   10                  15      
Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp 
            20                  25                  30          
Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys 
        35                  40                  45              
Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp 
    50                  55                  60                  
Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu 
65                  70                  75                  80  
Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn 
                85                  90                  95      
Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn 
            100                 105                 110         
Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu 
        115                 120                 125             
Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe 
    130                 135                 140                 
Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn 
145                 150                 155                 160 
Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile 
                165                 170                 175     
Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys 
            180                 185                 190         
Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys 
        195                 200                 205             
Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe 
    210                 215                 220                 
Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile 
225                 230                 235                 240 
Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn 
                245                 250                 255     
Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys 
            260                 265                 270         
Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser 
        275                 280                 285             
Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe 
    290                 295                 300                 
Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys 
305                 310                 315                 320 
Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile 
                325                 330                 335     
Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe 
            340                 345                 350         
Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp 
        355                 360                 365             
Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp 
    370                 375                 380                 
Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu 
385                 390                 395                 400 
Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu 
                405                 410                 415     
Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser 
            420                 425                 430         
Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys 
        435                 440                 445             
Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys 
    450                 455                 460                 
Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr 
465                 470                 475                 480 
Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile 
                485                 490                 495     
Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr 
            500                 505                 510         
Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro 
        515                 520                 525             
Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala 
    530                 535                 540                 
Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys 
545                 550                 555                 560 
Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly 
                565                 570                 575     
Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met 
            580                 585                 590         
Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro 
        595                 600                 605             
Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly 
    610                 615                 620                 
Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys 
625                 630                 635                 640 
Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn 
                645                 650                 655     
Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu 
            660                 665                 670         
Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys 
        675                 680                 685             
Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile 
    690                 695                 700                 
Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His 
705                 710                 715                 720 
Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile 
                725                 730                 735     
Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys 
            740                 745                 750         
Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys 
        755                 760                 765             
Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr 
    770                 775                 780                 
Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile 
785                 790                 795                 800 
Ala Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val 
                805                 810                 815     
Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Asp 
            820                 825                 830         
Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly 
        835                 840                 845             
Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn 
    850                 855                 860                 
Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu 
865                 870                 875                 880 
Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile 
                885                 890                 895     
Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 
            900                 905                 910         
Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu Asn 
        915                 920                 925             
Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr Gln 
    930                 935                 940                 
Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp Lys 
945                 950                 955                 960 
Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln Ile 
                965                 970                 975     
Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly Phe 
            980                 985                 990         
Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser Thr 
        995                 1000                1005            
Gly Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala Asp Ser 
    1010                1015                1020                
Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro Glu Glu 
1025                1030                1035                1040
Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser Arg Thr Asp 
                1045                1050                1055    
Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile 
            1060                1065                1070        
Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu 
        1075                1080                1085            
Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile 
    1090                1095                1100                
Asn Tyr Gln Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp 
1105                1110                1115                1120
Lys Ala Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln 
                1125                1130                1135    
Met Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 
            1140                1145                1150        
Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr Glu 
        1155                1160                1165            
Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala 
    1170                1175                1180                
Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys 
1185                1190                1195                1200
Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala Ile Ser Asn Lys 
                1205                1210                1215    
Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys His 
            1220                1225            

<210> 23
<211> 1368
<212> PRT
<213> Artificial Sequence

<220> 
<223> SpCas9

<400> 23
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         
Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe 
        995                 1000                1005            
Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys 
    1010                1015                1020                
Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser 
1025                1030                1035                1040
Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu 
                1045                1050                1055    
Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile 
            1060                1065                1070        
Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser 
        1075                1080                1085            
Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly 
    1090                1095                1100                
Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile 
1105                1110                1115                1120
Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser 
                1125                1130                1135    
Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly 
            1140                1145                1150        
Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile 
        1155                1160                1165            
Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala 
    1170                1175                1180                
Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys 
1185                1190                1195                1200
Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser 
                1205                1210                1215    
Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr 
            1220                1225                1230        
Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser 
        1235                1240                1245            
Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His 
    1250                1255                1260                
Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val 
1265                1270                1275                1280
Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys 
                1285                1290                1295    
His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu 
            1300                1305                1310        
Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp 
        1315                1320                1325            
Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp 
    1330                1335                1340                
Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile 
1345                1350                1355                1360
Asp Leu Ser Gln Leu Gly Gly Asp 
                1365            

<210> 24
<211> 1121
<212> PRT
<213> Artificial Sequence

<220> 
<223> St1Cas9

<400> 24
Met Ser Asp Leu Val Leu Gly Leu Asp Ile Gly Ile Gly Ser Val Gly 
1               5                   10                  15      
Val Gly Ile Leu Asn Lys Val Thr Gly Glu Ile Ile His Lys Asn Ser 
            20                  25                  30          
Arg Ile Phe Pro Ala Ala Gln Ala Glu Asn Asn Leu Val Arg Arg Thr 
        35                  40                  45              
Asn Arg Gln Gly Arg Arg Leu Ala Arg Arg Lys Lys His Arg Arg Val 
    50                  55                  60                  
Arg Leu Asn Arg Leu Phe Glu Glu Ser Gly Leu Ile Thr Asp Phe Thr 
65                  70                  75                  80  
Lys Ile Ser Ile Asn Leu Asn Pro Tyr Gln Leu Arg Val Lys Gly Leu 
                85                  90                  95      
Thr Asp Glu Leu Ser Asn Glu Glu Leu Phe Ile Ala Leu Lys Asn Met 
            100                 105                 110         
Val Lys His Arg Gly Ile Ser Tyr Leu Asp Asp Ala Ser Asp Asp Gly 
        115                 120                 125             
Asn Ser Ser Val Gly Asp Tyr Ala Gln Ile Val Lys Glu Asn Ser Lys 
    130                 135                 140                 
Gln Leu Glu Thr Lys Thr Pro Gly Gln Ile Gln Leu Glu Arg Tyr Gln 
145                 150                 155                 160 
Thr Tyr Gly Gln Leu Arg Gly Asp Phe Thr Val Glu Lys Asp Gly Lys 
                165                 170                 175     
Lys His Arg Leu Ile Asn Val Phe Pro Thr Ser Ala Tyr Arg Ser Glu 
            180                 185                 190         
Ala Leu Arg Ile Leu Gln Thr Gln Gln Glu Phe Asn Pro Gln Ile Thr 
        195                 200                 205             
Asp Glu Phe Ile Asn Arg Tyr Leu Glu Ile Leu Thr Gly Lys Arg Lys 
    210                 215                 220                 
Tyr Tyr His Gly Pro Gly Asn Glu Lys Ser Arg Thr Asp Tyr Gly Arg 
225                 230                 235                 240 
Tyr Arg Thr Ser Gly Glu Thr Leu Asp Asn Ile Phe Gly Ile Leu Ile 
                245                 250                 255     
Gly Lys Cys Thr Phe Tyr Pro Asp Glu Phe Arg Ala Ala Lys Ala Ser 
            260                 265                 270         
Tyr Thr Ala Gln Glu Phe Asn Leu Leu Asn Asp Leu Asn Asn Leu Thr 
        275                 280                 285             
Val Pro Thr Glu Thr Lys Lys Leu Ser Lys Glu Gln Lys Asn Gln Ile 
    290                 295                 300                 
Ile Asn Tyr Val Lys Asn Glu Lys Ala Met Gly Pro Ala Lys Leu Phe 
305                 310                 315                 320 
Lys Tyr Ile Ala Lys Leu Leu Ser Cys Asp Val Ala Asp Ile Lys Gly 
                325                 330                 335     
Tyr Arg Ile Asp Lys Ser Gly Lys Ala Glu Ile His Thr Phe Glu Ala 
            340                 345                 350         
Tyr Arg Lys Met Lys Thr Leu Glu Thr Leu Asp Ile Glu Gln Met Asp 
        355                 360                 365             
Arg Glu Thr Leu Asp Lys Leu Ala Tyr Val Leu Thr Leu Asn Thr Glu 
    370                 375                 380                 
Arg Glu Gly Ile Gln Glu Ala Leu Glu His Glu Phe Ala Asp Gly Ser 
385                 390                 395                 400 
Phe Ser Gln Lys Gln Val Asp Glu Leu Val Gln Phe Arg Lys Ala Asn 
                405                 410                 415     
Ser Ser Ile Phe Gly Lys Gly Trp His Asn Phe Ser Val Lys Leu Met 
            420                 425                 430         
Met Glu Leu Ile Pro Glu Leu Tyr Glu Thr Ser Glu Glu Gln Met Thr 
        435                 440                 445             
Ile Leu Thr Arg Leu Gly Lys Gln Lys Thr Thr Ser Ser Ser Asn Lys 
    450                 455                 460                 
Thr Lys Tyr Ile Asp Glu Lys Leu Leu Thr Glu Glu Ile Tyr Asn Pro 
465                 470                 475                 480 
Val Val Ala Lys Ser Val Arg Gln Ala Ile Lys Ile Val Asn Ala Ala 
                485                 490                 495     
Ile Lys Glu Tyr Gly Asp Phe Asp Asn Ile Val Ile Glu Met Ala Arg 
            500                 505                 510         
Glu Thr Asn Glu Asp Asp Glu Lys Lys Ala Ile Gln Lys Ile Gln Lys 
        515                 520                 525             
Ala Asn Lys Asp Glu Lys Asp Ala Ala Met Leu Lys Ala Ala Asn Gln 
    530                 535                 540                 
Tyr Asn Gly Lys Ala Glu Leu Pro His Ser Val Phe His Gly His Lys 
545                 550                 555                 560 
Gln Leu Ala Thr Lys Ile Arg Leu Trp His Gln Gln Gly Glu Arg Cys 
                565                 570                 575     
Leu Tyr Thr Gly Lys Thr Ile Ser Ile His Asp Leu Ile Asn Asn Ser 
            580                 585                 590         
Asn Gln Phe Glu Val Asp His Ile Leu Pro Leu Ser Ile Thr Phe Asp 
        595                 600                 605             
Asp Ser Leu Ala Asn Lys Val Leu Val Tyr Ala Thr Ala Asn Gln Glu 
    610                 615                 620                 
Lys Gly Gln Arg Thr Pro Tyr Gln Ala Leu Asp Ser Met Asp Asp Ala 
625                 630                 635                 640 
Trp Ser Phe Arg Glu Leu Lys Ala Phe Val Arg Glu Ser Lys Thr Leu 
                645                 650                 655     
Ser Asn Lys Lys Lys Glu Tyr Leu Leu Thr Glu Glu Asp Ile Ser Lys 
            660                 665                 670         
Phe Asp Val Arg Lys Lys Phe Ile Glu Arg Asn Leu Val Asp Thr Arg 
        675                 680                 685             
Tyr Ala Ser Arg Val Val Leu Asn Ala Leu Gln Glu His Phe Arg Ala 
    690                 695                 700                 
His Lys Ile Asp Thr Lys Val Ser Val Val Arg Gly Gln Phe Thr Ser 
705                 710                 715                 720 
Gln Leu Arg Arg His Trp Gly Ile Glu Lys Thr Arg Asp Thr Tyr His 
                725                 730                 735     
His His Ala Val Asp Ala Leu Ile Ile Ala Ala Ser Ser Gln Leu Asn 
            740                 745                 750         
Leu Trp Lys Lys Gln Lys Asn Thr Leu Val Ser Tyr Ser Glu Asp Gln 
        755                 760                 765             
Leu Leu Asp Ile Glu Thr Gly Glu Leu Ile Ser Asp Asp Glu Tyr Lys 
    770                 775                 780                 
Glu Ser Val Phe Lys Ala Pro Tyr Gln His Phe Val Asp Thr Leu Lys 
785                 790                 795                 800 
Ser Lys Glu Phe Glu Asp Ser Ile Leu Phe Ser Tyr Gln Val Asp Ser 
                805                 810                 815     
Lys Phe Asn Arg Lys Ile Ser Asp Ala Thr Ile Tyr Ala Thr Arg Gln 
            820                 825                 830         
Ala Lys Val Gly Lys Asp Lys Ala Asp Glu Thr Tyr Val Leu Gly Lys 
        835                 840                 845             
Ile Lys Asp Ile Tyr Thr Gln Asp Gly Tyr Asp Ala Phe Met Lys Ile 
    850                 855                 860                 
Tyr Lys Lys Asp Lys Ser Lys Phe Leu Met Tyr Arg His Asp Pro Gln 
865                 870                 875                 880 
Thr Phe Glu Lys Val Ile Glu Pro Ile Leu Glu Asn Tyr Pro Asn Lys 
                885                 890                 895     
Gln Ile Asn Glu Lys Gly Lys Glu Val Pro Cys Asn Pro Phe Leu Lys 
            900                 905                 910         
Tyr Lys Glu Glu His Gly Tyr Ile Arg Lys Tyr Ser Lys Lys Gly Asn 
        915                 920                 925             
Gly Pro Glu Ile Lys Ser Leu Lys Tyr Tyr Asp Ser Lys Leu Gly Asn 
    930                 935                 940                 
His Ile Asp Ile Thr Pro Lys Asp Ser Asn Asn Lys Val Val Leu Gln 
945                 950                 955                 960 
Ser Val Ser Pro Trp Arg Ala Asp Val Tyr Phe Asn Lys Thr Thr Gly 
                965                 970                 975     
Lys Tyr Glu Ile Leu Gly Leu Lys Tyr Ala Asp Leu Gln Phe Glu Lys 
            980                 985                 990         
Gly Thr Gly Thr Tyr Lys Ile Ser Gln Glu Lys Tyr Asn Asp Ile Lys 
        995                 1000                1005            
Lys Lys Glu Gly Val Asp Ser Asp Ser Glu Phe Lys Phe Thr Leu Tyr 
    1010                1015                1020                
Lys Asn Asp Leu Leu Leu Val Lys Asp Thr Glu Thr Lys Glu Gln Gln 
1025                1030                1035                1040
Leu Phe Arg Phe Leu Ser Arg Thr Met Pro Lys Gln Lys His Tyr Val 
                1045                1050                1055    
Glu Leu Lys Pro Tyr Asp Lys Gln Lys Phe Glu Gly Gly Glu Ala Leu 
            1060                1065                1070        
Ile Lys Val Leu Gly Asn Val Ala Asn Ser Gly Gln Cys Lys Lys Gly 
        1075                1080                1085            
Leu Gly Lys Ser Asn Ile Ser Ile Tyr Lys Val Arg Thr Asp Val Leu 
    1090                1095                1100                
Gly Asn Gln His Ile Ile Lys Asn Glu Gly Asp Lys Pro Lys Leu Asp 
1105                1110                1115                1120
Phe 
    

<210> 25
<211> 1053
<212> PRT
<213> Artificial Sequence

<220> 
<223> SaCas9

<400> 25
Met Lys Arg Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val 
1               5                   10                  15      
Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          
Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              
Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  
Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  
Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      
Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         
Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             
Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 
Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 
Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     
Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         
Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             
Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 
Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 
Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     
Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         
Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             
Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 
Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 
Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     
Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         
Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             
Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 
Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 
Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     
Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         
Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             
Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 
Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 
Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     
Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         
Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His Asp 
        515                 520                 525             
Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 
Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 
Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     
Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         
Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             
Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 
Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 
Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     
Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         
Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys Trp 
        675                 680                 685             
Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 
Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 
Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     
Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         
Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             
Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Glu Leu Ile 
    770                 775                 780                 
Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 
Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     
Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         
Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             
Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 
Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 
Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     
Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         
Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             
Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 
Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 
Glu Phe Ile Ala Ser Phe Tyr Asn Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     
Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile 
            980                 985                 990         
Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met 
        995                 1000                1005            
Asn Asp Lys Arg Pro Pro Arg Ile Ile Lys Thr Ile Ala Ser Lys Thr 
    1010                1015                1020                
Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr Glu 
1025                1030                1035                1040
Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly 
                1045                1050            

<210> 26
<211> 704
<212> DNA
<213> Artificial Sequence

<220> 
<223> frame-shift version of mNeonGreen

<400> 26
atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca tgagttacac      60

atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca     120

aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc     180

tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg     240

atgtcgcctt tccaggccgc catggtagat ggctccggat accaagtcca tcgcacaatg     300

cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac     360

atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc     420

aactcgctga ccgctgcgga cgcaggtcga agaagactta ccccaacgac aaaaccatca     480

tcagtacctt taagtggagt tacaccactg gaaatggcaa gcgctaccgg agcactgcgc     540

ggaccaccta cacctttgcc aagccaatgg cggctaacta tctgaagaac cagccgatgt     600

acgtgttccg taagacggag ctcaagcact ccaagaccga gctcaacttc aaggagtggc     660

aaaaggcctt taccgatgtg atgggcatgg acgagctgta caag                      704


<210> 27
<211> 708
<212> DNA
<213> Artificial Sequence

<220> 
<223> Coding region of mNeonGreen

<400> 27
atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca tgagttacac      60

atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca     120

aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc     180

tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg     240

atgtcgcctt tccaggccgc catggtagat ggctccggat accaagtcca tcgcacaatg     300

cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac     360

atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc     420

aactcgctga ccgctgcgga ctggtgcagg tcgaagaaga cttaccccaa cgacaaaacc     480

atcatcagta cctttaagtg gagttacacc actggaaatg gcaagcgcta ccggagcact     540

gcgcggacca cctacacctt tgccaagcca atggcggcta actatctgaa gaaccagccg     600

atgtacgtgt tccgtaagac ggagctcaag cactccaaga ccgagctcaa cttcaaggag     660

tggcaaaagg cctttaccga tgtgatgggc atggacgagc tgtacaag                  708


<210> 28
<211> 236
<212> PRT
<213> Artificial Sequence

<220> 
<223> mNeon Green

<400> 28
Met Val Ser Lys Gly Glu Glu Asp Asn Met Ala Ser Leu Pro Ala Thr 
1               5                   10                  15      
His Glu Leu His Ile Phe Gly Ser Ile Asn Gly Val Asp Phe Asp Met 
            20                  25                  30          
Val Gly Gln Gly Thr Gly Asn Pro Asn Asp Gly Tyr Glu Glu Leu Asn 
        35                  40                  45              
Leu Lys Ser Thr Lys Gly Asp Leu Gln Phe Ser Pro Trp Ile Leu Val 
    50                  55                  60                  
Pro His Ile Gly Tyr Gly Phe His Gln Tyr Leu Pro Tyr Pro Asp Gly 
65                  70                  75                  80  
Met Ser Pro Phe Gln Ala Ala Met Val Asp Gly Ser Gly Tyr Gln Val 
                85                  90                  95      
His Arg Thr Met Gln Phe Glu Asp Gly Ala Ser Leu Thr Val Asn Tyr 
            100                 105                 110         
Arg Tyr Thr Tyr Glu Gly Ser His Ile Lys Gly Glu Ala Gln Val Lys 
        115                 120                 125             
Gly Thr Gly Phe Pro Ala Asp Gly Pro Val Met Thr Asn Ser Leu Thr 
    130                 135                 140                 
Ala Ala Asp Trp Cys Arg Ser Lys Lys Thr Tyr Pro Asn Asp Lys Thr 
145                 150                 155                 160 
Ile Ile Ser Thr Phe Lys Trp Ser Tyr Thr Thr Gly Asn Gly Lys Arg 
                165                 170                 175     
Tyr Arg Ser Thr Ala Arg Thr Thr Tyr Thr Phe Ala Lys Pro Met Ala 
            180                 185                 190         
Ala Asn Tyr Leu Lys Asn Gln Pro Met Tyr Val Phe Arg Lys Thr Glu 
        195                 200                 205             
Leu Lys His Ser Lys Thr Glu Leu Asn Phe Lys Glu Trp Gln Lys Ala 
    210                 215                 220                 
Phe Thr Asp Val Met Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235     

<210> 29
<211> 704
<212> DNA
<213> Artificial Sequence

<220> 
<223> mutated coding region of mNeonGreen

<400> 29
atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca tgagttacac      60

atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca     120

aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc     180

tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg     240

atgtcgcctt tccaggccgc catggtagat ggctccggat accaagtcca tcgcacaatg     300

cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac     360

atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc     420

aactcgctga ccgctgcgga cgcaggtcga agaagactta ccccaacgac aaaaccatca     480

tcagtacctt taagtggagt tacaccactg gaaatggcaa gcgctaccgg agcactgcgc     540

ggaccaccta cacctttgcc aagccaatgg cggctaacta tctgaagaac cagccgatgt     600

acgtgttccg taagacggag ctcaagcact ccaagaccga gctcaacttc aaggagtggc     660

aaaaggcctt taccgatgtg atgggcatgg acgagctgta caag                      704


<210> 30
<211> 105
<212> DNA
<213> Artificial Sequence

<220> 
<223> Donor template NSFS-R

<400> 30
ggtttccctg ctgacggtcc tgtgatgacc aactcgctga ccgctgcgga ctggtgcagg     60

tcgaagaaga cttaccccaa cgacaaaacc atcatcagta ccttt                    105


<210> 31
<211> 237
<212> PRT
<213> Artificial Sequence

<220> 
<223> mRuby2

<400> 31
Met Val Ser Lys Gly Glu Glu Leu Ile Lys Glu Asn Met Arg Met Lys 
1               5                   10                  15      
Val Val Met Glu Gly Ser Val Asn Gly His Gln Phe Lys Cys Thr Gly 
            20                  25                  30          
Glu Gly Glu Gly Asn Pro Tyr Met Gly Thr Gln Thr Met Arg Ile Lys 
        35                  40                  45              
Val Ile Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr 
    50                  55                  60                  
Ser Phe Met Tyr Gly Ser Arg Thr Phe Ile Lys Tyr Pro Lys Gly Ile 
65                  70                  75                  80  
Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg 
                85                  90                  95      
Val Thr Arg Tyr Glu Asp Gly Gly Val Val Thr Val Met Gln Asp Thr 
            100                 105                 110         
Ser Leu Glu Asp Gly Cys Leu Val Tyr His Val Gln Val Arg Gly Val 
        115                 120                 125             
Asn Phe Pro Ser Asn Gly Pro Val Met Gln Lys Lys Thr Lys Gly Trp 
    130                 135                 140                 
Glu Pro Asn Thr Glu Met Met Tyr Pro Ala Asp Gly Gly Leu Arg Gly 
145                 150                 155                 160 
Tyr Thr His Met Ala Leu Lys Val Asp Gly Gly Gly His Leu Ser Cys 
                165                 170                 175     
Ser Phe Val Thr Thr Tyr Arg Ser Lys Lys Thr Val Gly Asn Ile Lys 
            180                 185                 190         
Met Pro Gly Ile His Ala Val Asp His Arg Leu Glu Arg Leu Glu Glu 
        195                 200                 205             
Ser Asp Asn Glu Met Phe Val Val Gln Arg Glu His Ala Val Ala Lys 
    210                 215                 220                 
Phe Ala Gly Leu Gly Gly Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235         

<210> 32
<211> 199
<212> PRT
<213> Artificial Sequence

<220> 
<223> Puromycin Resistance gene

<400> 32
Met Thr Glu Tyr Lys Pro Thr Val Arg Leu Ala Thr Arg Asp Asp Val 
1               5                   10                  15      
Pro Arg Ala Val Arg Thr Leu Ala Ala Ala Phe Ala Asp Tyr Pro Ala 
            20                  25                  30          
Thr Arg His Thr Val Asp Pro Asp Arg His Ile Glu Arg Val Thr Glu 
        35                  40                  45              
Leu Gln Glu Leu Phe Leu Thr Arg Val Gly Leu Asp Ile Gly Lys Val 
    50                  55                  60                  
Trp Val Ala Asp Asp Gly Ala Ala Val Ala Val Trp Thr Thr Pro Glu 
65                  70                  75                  80  
Ser Val Glu Ala Gly Ala Val Phe Ala Glu Ile Gly Pro Arg Met Ala 
                85                  90                  95      
Glu Leu Ser Gly Ser Arg Leu Ala Ala Gln Gln Gln Met Glu Gly Leu 
            100                 105                 110         
Leu Ala Pro His Arg Pro Lys Glu Pro Ala Trp Phe Leu Ala Thr Val 
        115                 120                 125             
Gly Val Ser Pro Asp His Gln Gly Lys Gly Leu Gly Ser Ala Val Val 
    130                 135                 140                 
Leu Pro Gly Val Glu Ala Ala Glu Arg Ala Gly Val Pro Ala Phe Leu 
145                 150                 155                 160 
Glu Thr Ser Ala Pro Arg Asn Leu Pro Phe Tyr Glu Arg Leu Gly Phe 
                165                 170                 175     
Thr Val Thr Ala Asp Val Glu Val Pro Glu Gly Pro Arg Thr Trp Cys 
            180                 185                 190         
Met Thr Arg Lys Pro Gly Ala 
        195                 

<210> 33
<211> 109
<212> DNA
<213> Artificial Sequence

<220> 
<223> Oligonucleotide for codon diversifaction of the mRuby2
      Chormophore Region

<220> 
<221> unsure
<222> 52..53
<223> /note="n = any nucleic acid"

<220> 
<221> unsure
<222> 55..56
<223> /note="n = any nucleic acid"

<220> 
<221> unsure
<222> 58..59
<223> /note="n = any nucleic acid"

<400> 33
tgtttaaaga aatcaggaat gcctttcggg tacttgataa aagtacggct vnnvnnvnng     60

aacgacgtgg caagaatgtc aaaggcaaat ggcaggggtc ctccctcga                109


<210> 34
<211> 117
<212> DNA
<213> Artificial Sequence

<220> 
<223> Oligo used for inducing a frame-shift near the chromophore region
      of mRuby2

<400> 34
agtcatcgag ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcgtatg     60

gcagccgtac ttttatcaag tacccgaaag gcattcctga tttctttaaa cagtcct       117


<210> 35
<211> 21
<212> PRT
<213> Artificial Sequence

<220> 
<223> Gene specific primer for RT PCR

<400> 35
Cys Thr Thr Gly Thr Ala Cys Ala Gly Cys Thr Cys Gly Thr Cys Cys 
1               5                   10                  15      
Ala Thr Cys Cys Cys 
            20      

<210> 36
<211> 42
<212> DNA
<213> Artificial Sequence

<220> 
<223> Deep sequencing primer 1

<400> 36
tacacgacgc tcttccgatc tatgcacagg tgaaggagaa gg                        42


<210> 37
<211> 42
<212> DNA
<213> Artificial Sequence

<220> 
<223> Deep sequencing primer 2

<400> 37
cagacgtgtg ctcttccgat cctccaccat cttcgtatct cg                        42


<210> 38
<211> 41
<212> DNA
<213> Artificial Sequence

<220> 
<223> Forward primer to extract the coding domains of the repaired
      mNeonGreen

<400> 38
ataaggatcc ggccaccatg gtgagcaagg gcgaggagga t                         41


<210> 39
<211> 42
<212> DNA
<213> Artificial Sequence

<220> 
<223> Reverse primer to extract the coding domains of the repaired
      mNeonGreen

<400> 39
tataggaatt cctattatca cttgtacagc tcgtccatgc cc                        42


<210> 40
<211> 31
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 1.F

<400> 40
ctttaagtgg acaccactgg aaatggcaag c                                   31


<210> 41
<211> 38
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 1.R

<400> 41
ccagtggtgt ccacttaaag gtactgatga tggttttg                             38


<210> 42
<211> 35
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame Shifting Primer for PCR Mutagenesis, 2.F

<400> 42
ctggtgcagg agaagactta ccccaacgac aaaac                                35


<210> 43
<211> 28
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 2.R

<400> 43
taagtcttct cctgcaccag tccgcagc                                       28


<210> 44
<211> 30
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 3.F

<400> 44
caggtgaagg tggtttccct gctgacggtc                                     30


<210> 45
<211> 29
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 3.R

<400> 45
agggaaacca ccttcacctg ggcctctcc                                      29


<210> 46
<211> 34
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 4.F

<400> 46
tcgggtatgg catcagtacc tgccctaccc tgac                                 34


<210> 47
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 4.R

<400> 47
ggtactgatg ccatacccga tatgagggac cag                                  33


<210> 48
<211> 25
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 5.F

<400> 48
gtccgcagcg gtcagcgagt tggtc                                          25


<210> 49
<211> 26
<212> DNA
<213> Artificial Sequence

<220> 
<223> Frame-Shifting Primer for PCR Mutagenesis, 5.R

<400> 49
gcaaccgtaa agttcaagta caaagg                                         26


<210> 50
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> PAM sequence for SaCas9

<400> 50
Asn Asn Gly Arg Arg Thr 
1               5       

<210> 51
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> PAM sequence for SaCas9

<400> 51
Asn Asn Gly Arg Arg Asn 
1               5       

<210> 52
<211> 7
<212> PRT
<213> Artificial Sequence

<220> 
<223> PAM sequence for St1Cas9

<400> 52
Asn Asn Ala Gly Ala Ala Trp 
1               5           

<210> 53
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 53
tcgctgaccg ctgcgggcag tcgcaggtcg aagaagact                            39


<210> 54
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 54
Ser Leu Thr Ala Ala Gly Ser Arg Arg Ser Lys Lys Thr 
1               5                   10              

<210> 55
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 55
tcgctgaccg ctgcgagtgt gtccaggtcg aagaagact                            39


<210> 56
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 56
Ser Leu Thr Ala Ala Ser Val Ser Arg Ser Lys Lys Thr 
1               5                   10              

<210> 57
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 57
tcgctgaccg ctgcggggtg ttgtaggtcg aagaagact                            39


<210> 58
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 58
Ser Leu Thr Ala Ala Gly Cys Cys Arg Ser Lys Lys Thr 
1               5                   10              

<210> 59
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 59
tcgctgaccg ctgcgatgat gggcaggtcg aagaagact                            39


<210> 60
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 60
Ser Leu Thr Ala Ala Met Met Gly Arg Ser Lys Lys Thr 
1               5                   10              

<210> 61
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 61
tcgctgaccg ctgcgtgtgt ctcgaggtcg aagaagact                            39


<210> 62
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 62
Ser Leu Thr Ala Ala Cys Val Ser Arg Ser Lys Lys Thr 
1               5                   10              

<210> 63
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 63
tcgctgaccg ctgcgggcgc caccaggtcg aagaagact                            39


<210> 64
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 64
Ser Leu Thr Ala Ala Gly Ala Thr Arg Ser Lys Lys Thr 
1               5                   10              

<210> 65
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 65
tcgctgaccg ctgcgaagtt ctctaggtcg aagaagact                            39


<210> 66
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 66
Ser Leu Thr Ala Ala Lys Phe Ser Arg Ser Lys Lys Thr 
1               5                   10              

<210> 67
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 67
tcgctgaccg ctgcgtcgct tatgaggtcg aagaagact                            39


<210> 68
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 68
Ser Leu Thr Ala Ala Ser Leu Met Arg Ser Lys Lys Thr 
1               5                   10              

<210> 69
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 69
tcgctgaccg ctgcgagtaa gcgcaggtcg aagaagact                            39


<210> 70
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 70
Ser Leu Thr Ala Ala Ser Lys Arg Arg Ser Lys Lys Thr 
1               5                   10              

<210> 71
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 71
tcgctgaccg ctgcgcaggc tactaggtcg aagaagact                            39


<210> 72
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 72
Ser Leu Thr Ala Ala Gln Ala Thr Arg Ser Lys Lys Thr 
1               5                   10              

<210> 73
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 73
tcgctgaccg ctgcgtgcgt gtgtaggtcg aagaagact                            39


<210> 74
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 74
Ser Leu Thr Ala Ala Cys Val Cys Arg Ser Lys Lys Thr 
1               5                   10              

<210> 75
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 75
tcgctgaccg ctgcgcggat ggggaggtcg aagaagact                            39


<210> 76
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 76
Ser Leu Thr Ala Ala Arg Met Gly Arg Ser Lys Lys Thr 
1               5                   10              

<210> 77
<211> 39
<212> DNA
<213> Artificial Sequence

<220> 
<223> DNA sequence of mNeonGreen variant

<400> 77
tcgctgaccg ctgcgatttg gacgaggtcg aagaagact                            39


<210> 78
<211> 13
<212> PRT
<213> Artificial Sequence

<220> 
<223> translated protein sequence of mNeonGreen variant

<400> 78
Ser Leu Thr Ala Ala Ile Trp Thr Arg Ser Lys Lys Thr 
1               5                   10              

<210> 79
<211> 219
<212> PRT
<213> Artificial Sequence

<220> 
<223> LanYFP

<400> 79
Met Ser Leu Pro Ala Thr His Glu Leu His Ile Phe Gly Ser Phe Asn 
1               5                   10                  15      
Gly Val Asp Phe Asp Met Val Gly Arg Gly Thr Gly Asn Pro Asn Asp 
            20                  25                  30          
Gly Tyr Glu Glu Leu Asn Leu Lys Ser Thr Lys Gly Ala Leu Gln Phe 
        35                  40                  45              
Ser Pro Trp Ile Leu Val Pro Gln Ile Gly Tyr Gly Phe His Gln Tyr 
    50                  55                  60                  
Leu Pro Phe Pro Asp Gly Met Ser Pro Phe Gln Ala Ala Met Lys Asp 
65                  70                  75                  80  
Gly Ser Gly Tyr Gln Val His Arg Thr Met Gln Phe Glu Asp Gly Ala 
                85                  90                  95      
Ser Leu Thr Ser Asn Tyr Arg Tyr Thr Tyr Glu Gly Ser His Ile Lys 
            100                 105                 110         
Gly Glu Phe Gln Val Ile Gly Thr Gly Phe Pro Ala Asp Gly Pro Val 
        115                 120                 125             
Met Thr Asn Ser Leu Thr Ala Ala Asp Trp Cys Val Thr Lys Met Leu 
    130                 135                 140                 
Tyr Pro Asn Asp Lys Thr Ile Ile Ser Thr Phe Asp Trp Thr Tyr Thr 
145                 150                 155                 160 
Thr Gly Ser Gly Lys Arg Tyr Gln Ser Thr Val Arg Thr Asn Tyr Thr 
                165                 170                 175     
Phe Ala Lys Pro Met Ala Ala Asn Ile Leu Lys Asn Gln Pro Met Phe 
            180                 185                 190         
Val Phe Arg Lys Thr Glu Leu Lys His Ser Lys Thr Glu Leu Asn Phe 
        195                 200                 205             
Lys Glu Trp Gln Lys Ala Phe Thr Asp Val Met 
    210                 215                 

<210> 80
<211> 236
<212> PRT
<213> Artificial Sequence

<220> 
<223> dLanYFP

<400> 80
Met Val Ser Lys Gly Glu Glu Asp Asn Met Ala Ser Leu Pro Ala Thr 
1               5                   10                  15      
His Glu Leu His Ile Phe Gly Ser Phe Asn Gly Val Asp Phe Asp Met 
            20                  25                  30          
Val Gly Arg Gly Thr Gly Asn Pro Asn Asp Gly Tyr Glu Glu Leu Asn 
        35                  40                  45              
Leu Lys Ser Thr Lys Gly Asp Leu Gln Phe Ser Pro Trp Ile Leu Val 
    50                  55                  60                  
Pro Gln Ile Gly Tyr Gly Phe His Gln Tyr Leu Pro Phe Pro Asp Gly 
65                  70                  75                  80  
Met Ser Pro Phe Gln Ala Ala Met Lys Asp Gly Ser Gly Tyr Gln Val 
                85                  90                  95      
His Arg Thr Met Gln Phe Glu Asp Gly Ala Ser Leu Thr Ser Asn Tyr 
            100                 105                 110         
Arg Tyr Thr Tyr Glu Gly Ser His Ile Lys Gly Glu Phe Gln Val Lys 
        115                 120                 125             
Gly Thr Gly Phe Pro Ala Asp Gly Pro Val Met Thr Asn Ser Leu Thr 
    130                 135                 140                 
Ala Ala Asp Trp Cys Val Thr Lys Met Leu Tyr Pro Asn Asp Lys Thr 
145                 150                 155                 160 
Ile Ile Ser Thr Phe Asp Trp Thr Tyr Thr Thr Gly Asn Gly Lys Arg 
                165                 170                 175     
Tyr Gln Ser Thr Ala Arg Thr Thr Tyr Thr Phe Ala Lys Pro Met Ala 
            180                 185                 190         
Ala Asn Ile Leu Lys Asn Gln Pro Met Phe Val Phe Arg Lys Thr Glu 
        195                 200                 205             
Leu Lys His Ser Lys Thr Glu Leu Asn Phe Lys Glu Trp Gln Lys Ala 
    210                 215                 220                 
Phe Thr Asp Val Met Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235     

<210> 81
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 1.F

<400> 81
caccgcatca tcagtacctt taag                                           24


<210> 82
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 1.R

<400> 82
aaaccttaaa ggtactgatg atgc                                           24


<210> 83
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 2.F

<400> 83
gtggcctggc gacgcctgac cacg                                           24


<210> 84
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 2.R

<400> 84
aaaccgtggt caggcgtcgc cagg                                           24


<210> 85
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 3.F

<400> 85
caccgaaagg agaggcccag gtga                                           24


<210> 86
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 3.R

<400> 86
aaactcacct gggcctctcc tttc                                           24


<210> 87
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 4.F

<400> 87
caccgtggtc cctcatatcg ggta                                           24


<210> 88
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 4.R

<400> 88
aaactacccg atatgaggga ccac                                           24


<210> 89
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 5.F

<400> 89
caccgcgctg accgctgcgg acgc                                           24


<210> 90
<211> 24
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer 5.R

<400> 90
aaacgcgtcc gcagcggtca gcgc                                           24


<210> 91
<211> 708
<212> DNA
<213> Artificial Sequence

<220> 
<223> mNeonGreen2

<400> 91
atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca tgagttacac      60

atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca     120

aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc     180

tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg     240

atgtcgcctt tccaggccgc catggtagat ggctccggat accaagtcca tcgcacaatg     300

cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac     360

atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc     420

aactcgctga ccgctgcgga tgcgtgctgg tcgaagaaga cttaccccaa cgacaaaacc     480

atcatcagta cctttaagtg gagttacacc actggaaatg gcaagcgcta ccggagcact     540

gcgcggacca cctacacctt tgccaagcca atggcggcta actatctgaa gaaccagccg     600

atgtacgtgt tccgtaagac ggagctcaag cactccaaga ccgagctcaa cttcaaggag     660

tggcaaaagg cctttaccga tgtgatgggc atggacgagc tgtacaag                  708


<210> 92
<211> 236
<212> PRT
<213> Artificial Sequence

<220> 
<223> mNeonGreen2

<400> 92
Met Val Ser Lys Gly Glu Glu Asp Asn Met Ala Ser Leu Pro Ala Thr 
1               5                   10                  15      
His Glu Leu His Ile Phe Gly Ser Ile Asn Gly Val Asp Phe Asp Met 
            20                  25                  30          
Val Gly Gln Gly Thr Gly Asn Pro Asn Asp Gly Tyr Glu Glu Leu Asn 
        35                  40                  45              
Leu Lys Ser Thr Lys Gly Asp Leu Gln Phe Ser Pro Trp Ile Leu Val 
    50                  55                  60                  
Pro His Ile Gly Tyr Gly Phe His Gln Tyr Leu Pro Tyr Pro Asp Gly 
65                  70                  75                  80  
Met Ser Pro Phe Gln Ala Ala Met Val Asp Gly Ser Gly Tyr Gln Val 
                85                  90                  95      
His Arg Thr Met Gln Phe Glu Asp Gly Ala Ser Leu Thr Val Asn Tyr 
            100                 105                 110         
Arg Tyr Thr Tyr Glu Gly Ser His Ile Lys Gly Glu Ala Gln Val Lys 
        115                 120                 125             
Gly Thr Gly Phe Pro Ala Asp Gly Pro Val Met Thr Asn Ser Leu Thr 
    130                 135                 140                 
Ala Ala Asp Ala Cys Trp Ser Lys Lys Thr Tyr Pro Asn Asp Lys Thr 
145                 150                 155                 160 
Ile Ile Ser Thr Phe Lys Trp Ser Tyr Thr Thr Gly Asn Gly Lys Arg 
                165                 170                 175     
Tyr Arg Ser Thr Ala Arg Thr Thr Tyr Thr Phe Ala Lys Pro Met Ala 
            180                 185                 190         
Ala Asn Tyr Leu Lys Asn Gln Pro Met Tyr Val Phe Arg Lys Thr Glu 
        195                 200                 205             
Leu Lys His Ser Lys Thr Glu Leu Asn Phe Lys Glu Trp Gln Lys Ala 
    210                 215                 220                 
Phe Thr Asp Val Met Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235     

<210> 93
<211> 4
<212> PRT
<213> Artificial Sequence

<220> 
<223> Amino acid within mNeonGreen2

<400> 93
Asp Ala Cys Trp 
1               

<210> 94
<211> 711
<212> PRT
<213> Artificial Sequence

<220> 
<223> mRuby2-TagBFP2-Puromycin

<400> 94
Met Val Ser Lys Gly Glu Glu Leu Ile Lys Glu Asn Met Arg Met Lys 
1               5                   10                  15      
Val Val Met Glu Gly Ser Val Asn Gly His Gln Phe Lys Cys Thr Gly 
            20                  25                  30          
Glu Gly Glu Gly Asn Pro Tyr Met Gly Thr Gln Thr Met Arg Ile Lys 
        35                  40                  45              
Val Ile Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr 
    50                  55                  60                  
Ser Phe Met Tyr Gly Ser Arg Thr Phe Ile Lys Tyr Pro Lys Gly Ile 
65                  70                  75                  80  
Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg 
                85                  90                  95      
Val Thr Arg Tyr Glu Asp Gly Gly Val Val Thr Val Met Gln Asp Thr 
            100                 105                 110         
Ser Leu Glu Asp Gly Cys Leu Val Tyr His Val Gln Val Arg Gly Val 
        115                 120                 125             
Asn Phe Pro Ser Asn Gly Pro Val Met Gln Lys Lys Thr Lys Gly Trp 
    130                 135                 140                 
Glu Pro Asn Thr Glu Met Met Tyr Pro Ala Asp Gly Gly Leu Arg Gly 
145                 150                 155                 160 
Tyr Thr His Met Ala Leu Lys Val Asp Gly Gly Gly His Leu Ser Cys 
                165                 170                 175     
Ser Phe Val Thr Thr Tyr Arg Ser Lys Lys Thr Val Gly Asn Ile Lys 
            180                 185                 190         
Met Pro Gly Ile His Ala Val Asp His Arg Leu Glu Arg Leu Glu Glu 
        195                 200                 205             
Ser Asp Asn Glu Met Phe Val Val Gln Arg Glu His Ala Val Ala Lys 
    210                 215                 220                 
Phe Ala Gly Leu Gly Gly Gly Met Asp Glu Leu Tyr Lys Ala Glu Ala 
225                 230                 235                 240 
Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Val Ser 
                245                 250                 255     
Lys Gly Glu Glu Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr Met 
            260                 265                 270         
Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr Ser Glu Gly Glu 
        275                 280                 285             
Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu 
    290                 295                 300                 
Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu 
305                 310                 315                 320 
Tyr Gly Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe 
                325                 330                 335     
Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr 
            340                 345                 350         
Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln 
        355                 360                 365             
Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe Thr 
    370                 375                 380                 
Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe 
385                 390                 395                 400 
Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp 
                405                 410                 415     
Met Ala Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ala Lys 
            420                 425                 430         
Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly 
        435                 440                 445             
Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn 
    450                 455                 460                 
Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys Asp 
465                 470                 475                 480 
Leu Pro Ser Lys Leu Gly His Lys Leu Asn Gly Ser Gly Ala Thr Asn 
                485                 490                 495     
Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro 
            500                 505                 510         
Met Thr Glu Tyr Lys Pro Thr Val Arg Leu Ala Thr Arg Asp Asp Val 
        515                 520                 525             
Pro Arg Ala Val Arg Thr Leu Ala Ala Ala Phe Ala Asp Tyr Pro Ala 
    530                 535                 540                 
Thr Arg His Thr Val Asp Pro Asp Arg His Ile Glu Arg Val Thr Glu 
545                 550                 555                 560 
Leu Gln Glu Leu Phe Leu Thr Arg Val Gly Leu Asp Ile Gly Lys Val 
                565                 570                 575     
Trp Val Ala Asp Asp Gly Ala Ala Val Ala Val Trp Thr Thr Pro Glu 
            580                 585                 590         
Ser Val Glu Ala Gly Ala Val Phe Ala Glu Ile Gly Pro Arg Met Ala 
        595                 600                 605             
Glu Leu Ser Gly Ser Arg Leu Ala Ala Gln Gln Gln Met Glu Gly Leu 
    610                 615                 620                 
Leu Ala Pro His Arg Pro Lys Glu Pro Ala Trp Phe Leu Ala Thr Val 
625                 630                 635                 640 
Gly Val Ser Pro Asp His Gln Gly Lys Gly Leu Gly Ser Ala Val Val 
                645                 650                 655     
Leu Pro Gly Val Glu Ala Ala Glu Arg Ala Gly Val Pro Ala Phe Leu 
            660                 665                 670         
Glu Thr Ser Ala Pro Arg Asn Leu Pro Phe Tyr Glu Arg Leu Gly Phe 
        675                 680                 685             
Thr Val Thr Ala Asp Val Glu Val Pro Glu Gly Pro Arg Thr Trp Cys 
    690                 695                 700                 
Met Thr Arg Lys Pro Gly Ala 
705                 710     

<210> 95
<211> 250
<212> DNA
<213> Artificial Sequence

<220> 
<223> sequenced region within the parental original mRuby2 sequence

<400> 95
ccaccaattc aaatgcacag gtgaaggaga aggcaatccg tacatgggaa ctcaaaccat     60

gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc    120

gttcatgtat ggcagccgta cttttatcaa gtacccgaaa ggcattcctg atttctttaa    180

acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt    240

cgtcaccgtc                                                           250


