
SEQUENCE LISTING


<110> CELLECTIS

<120> Engineering mammalian genome using DNA-guided Argonaute 
      interference systems (DAIS)

<130> P81401684PCT00

<150> PA201470141
<151> 2014-03-21

<160> 19

<170> PatentIn version 3.5


<210>  1

<211>  685
<212>  PRT
<213>  Thermus thermophilus
<220>

<400>  1

Met Asn His Leu Gly Lys Thr Glu Val Phe Leu Asn Arg Phe Ala Leu 
1               5                   10                  15      


Arg Pro Leu Asn Pro Glu Glu Leu Arg Pro Trp Arg Leu Glu Val Val 
            20                  25                  30          


Leu Asp Pro Pro Pro Gly Arg Glu Glu Val Tyr Pro Leu Leu Ala Gln 
        35                  40                  45              


Val Ala Arg Arg Ala Gly Gly Val Thr Val Arg Met Gly Asp Gly Leu 
    50                  55                  60                  


Ala Ser Trp Ser Pro Pro Glu Val Leu Val Leu Glu Gly Thr Leu Ala 
65                  70                  75                  80  


Arg Met Gly Gln Thr Tyr Ala Tyr Arg Leu Tyr Pro Lys Gly Arg Arg 
                85                  90                  95      


Pro Leu Asp Pro Lys Asp Pro Gly Glu Arg Ser Val Leu Ser Ala Leu 
            100                 105                 110         


Ala Arg Arg Leu Leu Gln Glu Arg Leu Arg Arg Leu Glu Gly Val Trp 
        115                 120                 125             


Val Glu Gly Leu Ala Val Tyr Arg Arg Glu His Ala Arg Gly Pro Gly 
    130                 135                 140                 


Trp Arg Val Leu Gly Gly Ala Val Leu Asp Leu Trp Val Ser Asp Ser 
145                 150                 155                 160 


Gly Ala Phe Leu Leu Glu Val Asp Pro Ala Tyr Arg Ile Leu Cys Glu 
                165                 170                 175     


Met Ser Leu Glu Ala Trp Leu Ala Gln Gly His Pro Leu Pro Lys Arg 
            180                 185                 190         


Val Arg Asn Ala Tyr Asp Arg Arg Thr Trp Glu Leu Leu Arg Leu Gly 
        195                 200                 205             


Glu Glu Asp Pro Lys Glu Leu Pro Leu Pro Gly Gly Leu Ser Leu Leu 
    210                 215                 220                 


Asp Tyr His Ala Ser Lys Gly Arg Leu Gln Gly Arg Glu Gly Gly Arg 
225                 230                 235                 240 


Val Ala Trp Val Ala Asp Pro Lys Asp Pro Arg Lys Pro Ile Pro His 
                245                 250                 255     


Leu Thr Gly Leu Leu Val Pro Val Leu Thr Leu Glu Asp Leu His Glu 
            260                 265                 270         


Glu Glu Gly Ser Leu Ala Leu Ser Leu Pro Trp Glu Glu Arg Arg Arg 
        275                 280                 285             


Arg Thr Arg Glu Ile Ala Ser Trp Ile Gly Arg Arg Leu Gly Leu Gly 
    290                 295                 300                 


Thr Pro Glu Ala Val Arg Ala Gln Ala Tyr Arg Leu Ser Ile Pro Lys 
305                 310                 315                 320 


Leu Met Gly Arg Arg Ala Val Ser Lys Pro Ala Asp Ala Leu Arg Val 
                325                 330                 335     


Gly Phe Tyr Arg Ala Gln Glu Thr Ala Leu Ala Leu Leu Arg Leu Asp 
            340                 345                 350         


Gly Ala Gln Gly Trp Pro Glu Phe Leu Arg Arg Ala Leu Leu Arg Ala 
        355                 360                 365             


Phe Gly Ala Ser Gly Ala Ser Leu Arg Leu His Thr Leu His Ala His 
    370                 375                 380                 


Pro Ser Gln Gly Leu Ala Phe Arg Glu Ala Leu Arg Lys Ala Lys Glu 
385                 390                 395                 400 


Glu Gly Val Gln Ala Val Leu Val Leu Thr Pro Pro Met Ala Trp Glu 
                405                 410                 415     


Asp Arg Asn Arg Leu Lys Ala Leu Leu Leu Arg Glu Gly Leu Pro Ser 
            420                 425                 430         


Gln Ile Leu Asn Val Pro Leu Arg Glu Glu Glu Arg His Arg Trp Glu 
        435                 440                 445             


Asn Ala Leu Leu Gly Leu Leu Ala Lys Ala Gly Leu Gln Val Val Ala 
    450                 455                 460                 


Leu Ser Gly Ala Tyr Pro Ala Glu Leu Ala Val Gly Phe Asp Ala Gly 
465                 470                 475                 480 


Gly Arg Glu Ser Phe Arg Phe Gly Gly Ala Ala Cys Ala Val Gly Gly 
                485                 490                 495     


Asp Gly Gly His Leu Leu Trp Thr Leu Pro Glu Ala Gln Ala Gly Glu 
            500                 505                 510         


Arg Ile Pro Gln Glu Val Val Trp Asp Leu Leu Glu Glu Thr Leu Trp 
        515                 520                 525             


Ala Phe Arg Arg Lys Ala Gly Arg Leu Pro Ser Arg Val Leu Leu Leu 
    530                 535                 540                 


Arg Asp Gly Arg Val Pro Gln Asp Glu Phe Ala Leu Ala Leu Glu Ala 
545                 550                 555                 560 


Leu Ala Arg Glu Gly Ile Ala Tyr Asp Leu Val Ser Val Arg Lys Ser 
                565                 570                 575     


Gly Gly Gly Arg Val Tyr Pro Val Gln Gly Arg Leu Ala Asp Gly Leu 
            580                 585                 590         


Tyr Val Pro Leu Glu Asp Lys Thr Phe Leu Leu Leu Thr Val His Arg 
        595                 600                 605             


Asp Phe Arg Gly Thr Pro Arg Pro Leu Lys Leu Val His Glu Ala Gly 
    610                 615                 620                 


Asp Thr Pro Leu Glu Ala Leu Ala His Gln Ile Phe His Leu Thr Arg 
625                 630                 635                 640 


Leu Tyr Pro Ala Ser Gly Phe Ala Phe Pro Arg Leu Pro Ala Pro Leu 
                645                 650                 655     


His Leu Ala Asp Arg Leu Val Lys Glu Val Gly Arg Leu Gly Ile Arg 
            660                 665                 670         


His Leu Lys Glu Val Asp Arg Glu Lys Leu Phe Phe Val 
        675                 680                 685 


<210>  2

<211>  42
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC target 1

<400>  2
cacaaagtaa ggattctgat gtgtatatca cagacaaaac tg                        42


<210>  3

<211>  42
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC target 2

<400>  3
catgaggtct atggacttca agagcaacag tgctgtggcc tg                        42


<210>  4

<211>  42
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC target 3

<400>  4
caaagtaagg attctgatgt gtatatcaca gacaaaactg tg                        42


<210>  5

<211>  22
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC target 7

<400>  5
ccacagatat ccagaaccct ga                                              22


<210>  6

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo a target 1

<400>  6
cacaaagtaa ggattctgat g                                               21


<210>  7

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 1

<400>  7
cagttttgtc tgtgatatac a                                               21


<210>  8

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo a target 2

<400>  8
catgaggtct atggacttca a                                               21


<210>  9

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 2

<400>  9
caggccacag cactgttgct c                                               21


<210>  10

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo a target 3

<400>  10
caaagtaagg attctgatgt g                                               21


<210>  11

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 3

<400>  11
cacagttttg tctgtgatat a                                               21


<210>  12

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo a target 7

<400>  12
cacagatatc cagaaccctg a                                               21


<210>  13

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 7a

<400>  13
cagggttctg gatatctgtg g                                               21


<210>  14

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 7b


<400>    
cagctggtac acggcagggt c                                            21


<210>  15

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 7c


<400>    
cactggattt agagtctctc a                                           21


<210>  16

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 7d


<400>    
cacggcaggg tcagggttct g                                          21


<210>  17

<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  description of artificial sequence: synthetic polynucleotide

<220>
<223>  TRAC Oligo b target 7e


<400>    
cagggtcagg gttctggata t                                         21


<210>  18
<211>  2820
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DAIS-2A-BFP

<400>  18
atgaatcacc tgggcaagac cgaggtgttc ctgaacaggt tcgccctcag gccactgaac       60

cccgaggagc tccggccctg gaggctcgaa gtggtgctcg acccaccccc tggaagagag      120

gaggtgtacc ccctgctcgc ccaggtcgca aggagagccg gaggagtgac cgtcagaatg      180

ggggacggac tggcatcctg gagcccacct gaggtgctgg tgctggaggg aaccctggca      240

agaatggggc agacctacgc ctacaggctc taccccaagg ggcgcagacc cctcgaccct      300

aaggaccctg gggagagaag cgtgctctcc gcactcgcaa ggagactgct ccaggagcgg      360

ctcaggagac tggagggagt gtgggtggag ggactcgcag tctacaggag ggagcacgca      420

aggggccctg gatggagagt cctcggagga gcagtgctcg acctctgggt gtccgattcc      480

ggggcattcc tgctggaggt ggaccctgct taccgcatcc tgtgcgagat gtccctggag      540

gcctggctcg cccagggaca ccctctccct aagcgggtga gaaacgccta cgaccggaga      600

acctgggagc tcctccggct gggagaggaa gatcccaagg agctccccct gcctggaggg      660

ctcagcctcc tcgactacca cgcttccaag gggagactgc aggggagaga ggggggcaga      720

gtcgcttggg tcgctgaccc taaagaccct agaaagccaa tcccccacct caccgggctg      780

ctggtgccag tcctgaccct ggaggacctc cacgaggagg aaggctctct ggctctcagc      840

ctcccttggg aggagaggcg ccgcagaaca agggagatcg cctcctggat cggcaggagg      900

cttggccttg ggacaccaga ggctgtgagg gctcaggctt accggctctc catcccaaag      960

ctgatggggc ggagggctgt gagcaagccc gctgacgctc tgagggtcgg gttctacagg     1020

gctcaggaaa cagccctggc cctcctgagg ctggacgggg ctcaagggtg gccagagttc     1080

ctgaggaggg ccctgctgag ggctttcggc gcttctggcg cctcacttag gctgcacaca     1140

ctgcacgccc acccatctca gggccttgcc ttcagggagg ccttgaggaa ggccaaggag     1200

gagggggtgc aggccgtcct tgtcttgact cctcctatgg cctgggagga caggaacagg     1260

ctgaaggccc tgcttcttag ggaggggttg ccaagccaga ttctgaacgt gcccctgcgg     1320

gaggaagaac ggcatcggtg ggagaacgcc ctgctgggct tgctggccaa agccgggctt     1380

caagtggtgg ccttgagcgg ggcctatcca gccgaactgg ccgtcgggtt tgatgccggc     1440

ggccgggaat catttcgctt cggcggggcc gcctgtgccg tgggggggga tggcggccat     1500

ttgttgtgga cacttccaga agcccaggcc ggcgaacgga ttccacagga agtcgtgtgg     1560

gatttgcttg aggagactct gtgggccttc cgccggaaag ccggccggtt gcccagtcgg     1620

gtcttgttgc ttcgggatgg gcgggtgcca caggatgagt ttgccttggc cttggaagcc     1680

cttgcccgcg aggggattgc ctatgatctg gtgagcgtgc ggaagagcgg gggcggccgg     1740

gtctaccccg tccagggccg gttggccgat ggcttgtacg tgcccttgga ggataaaact     1800

ttcttgctgc tgacagtgca ccgggatttt cggggcactc cccgccccct gaagctggtc     1860

cacgaggccg gcgatacacc cctggaggcc ctggcccatc agatttttca cctgactcgg     1920

ctgtaccccg ccagtggctt tgcctttccc cggctgcccg cccccctgca tctggccgat     1980

cggctggtca aggaggtcgg ccggctgggc atccggcatc tgaaggaggt ggatcgggag     2040

aagctgtttt ttgtgggatc cgagggcaga ggcagcctgc tgacctgcgg cgacgtggag     2100

gagaacccag gcccctccgg aagcgagctg attaaggaga acatgcacat gaagctgtac     2160

atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc     2220

tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc     2280

ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag     2340

ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc     2400

acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc     2460

ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag     2520

aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg     2580

gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc     2640

aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat     2700

gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac     2760

gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa gctcaattaa     2820


<210>  19
<211>  939
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  DAIS-2A-BFP

<400>  19

Met Asn His Leu Gly Lys Thr Glu Val Phe Leu Asn Arg Phe Ala Leu 
1               5                   10                  15      


Arg Pro Leu Asn Pro Glu Glu Leu Arg Pro Trp Arg Leu Glu Val Val 
            20                  25                  30          


Leu Asp Pro Pro Pro Gly Arg Glu Glu Val Tyr Pro Leu Leu Ala Gln 
        35                  40                  45              


Val Ala Arg Arg Ala Gly Gly Val Thr Val Arg Met Gly Asp Gly Leu 
    50                  55                  60                  


Ala Ser Trp Ser Pro Pro Glu Val Leu Val Leu Glu Gly Thr Leu Ala 
65                  70                  75                  80  


Arg Met Gly Gln Thr Tyr Ala Tyr Arg Leu Tyr Pro Lys Gly Arg Arg 
                85                  90                  95      


Pro Leu Asp Pro Lys Asp Pro Gly Glu Arg Ser Val Leu Ser Ala Leu 
            100                 105                 110         


Ala Arg Arg Leu Leu Gln Glu Arg Leu Arg Arg Leu Glu Gly Val Trp 
        115                 120                 125             


Val Glu Gly Leu Ala Val Tyr Arg Arg Glu His Ala Arg Gly Pro Gly 
    130                 135                 140                 


Trp Arg Val Leu Gly Gly Ala Val Leu Asp Leu Trp Val Ser Asp Ser 
145                 150                 155                 160 


Gly Ala Phe Leu Leu Glu Val Asp Pro Ala Tyr Arg Ile Leu Cys Glu 
                165                 170                 175     


Met Ser Leu Glu Ala Trp Leu Ala Gln Gly His Pro Leu Pro Lys Arg 
            180                 185                 190         


Val Arg Asn Ala Tyr Asp Arg Arg Thr Trp Glu Leu Leu Arg Leu Gly 
        195                 200                 205             


Glu Glu Asp Pro Lys Glu Leu Pro Leu Pro Gly Gly Leu Ser Leu Leu 
    210                 215                 220                 


Asp Tyr His Ala Ser Lys Gly Arg Leu Gln Gly Arg Glu Gly Gly Arg 
225                 230                 235                 240 


Val Ala Trp Val Ala Asp Pro Lys Asp Pro Arg Lys Pro Ile Pro His 
                245                 250                 255     


Leu Thr Gly Leu Leu Val Pro Val Leu Thr Leu Glu Asp Leu His Glu 
            260                 265                 270         


Glu Glu Gly Ser Leu Ala Leu Ser Leu Pro Trp Glu Glu Arg Arg Arg 
        275                 280                 285             


Arg Thr Arg Glu Ile Ala Ser Trp Ile Gly Arg Arg Leu Gly Leu Gly 
    290                 295                 300                 


Thr Pro Glu Ala Val Arg Ala Gln Ala Tyr Arg Leu Ser Ile Pro Lys 
305                 310                 315                 320 


Leu Met Gly Arg Arg Ala Val Ser Lys Pro Ala Asp Ala Leu Arg Val 
                325                 330                 335     


Gly Phe Tyr Arg Ala Gln Glu Thr Ala Leu Ala Leu Leu Arg Leu Asp 
            340                 345                 350         


Gly Ala Gln Gly Trp Pro Glu Phe Leu Arg Arg Ala Leu Leu Arg Ala 
        355                 360                 365             


Phe Gly Ala Ser Gly Ala Ser Leu Arg Leu His Thr Leu His Ala His 
    370                 375                 380                 


Pro Ser Gln Gly Leu Ala Phe Arg Glu Ala Leu Arg Lys Ala Lys Glu 
385                 390                 395                 400 


Glu Gly Val Gln Ala Val Leu Val Leu Thr Pro Pro Met Ala Trp Glu 
                405                 410                 415     


Asp Arg Asn Arg Leu Lys Ala Leu Leu Leu Arg Glu Gly Leu Pro Ser 
            420                 425                 430         


Gln Ile Leu Asn Val Pro Leu Arg Glu Glu Glu Arg His Arg Trp Glu 
        435                 440                 445             


Asn Ala Leu Leu Gly Leu Leu Ala Lys Ala Gly Leu Gln Val Val Ala 
    450                 455                 460                 


Leu Ser Gly Ala Tyr Pro Ala Glu Leu Ala Val Gly Phe Asp Ala Gly 
465                 470                 475                 480 


Gly Arg Glu Ser Phe Arg Phe Gly Gly Ala Ala Cys Ala Val Gly Gly 
                485                 490                 495     


Asp Gly Gly His Leu Leu Trp Thr Leu Pro Glu Ala Gln Ala Gly Glu 
            500                 505                 510         


Arg Ile Pro Gln Glu Val Val Trp Asp Leu Leu Glu Glu Thr Leu Trp 
        515                 520                 525             


Ala Phe Arg Arg Lys Ala Gly Arg Leu Pro Ser Arg Val Leu Leu Leu 
    530                 535                 540                 


Arg Asp Gly Arg Val Pro Gln Asp Glu Phe Ala Leu Ala Leu Glu Ala 
545                 550                 555                 560 


Leu Ala Arg Glu Gly Ile Ala Tyr Asp Leu Val Ser Val Arg Lys Ser 
                565                 570                 575     


Gly Gly Gly Arg Val Tyr Pro Val Gln Gly Arg Leu Ala Asp Gly Leu 
            580                 585                 590         


Tyr Val Pro Leu Glu Asp Lys Thr Phe Leu Leu Leu Thr Val His Arg 
        595                 600                 605             


Asp Phe Arg Gly Thr Pro Arg Pro Leu Lys Leu Val His Glu Ala Gly 
    610                 615                 620                 


Asp Thr Pro Leu Glu Ala Leu Ala His Gln Ile Phe His Leu Thr Arg 
625                 630                 635                 640 


Leu Tyr Pro Ala Ser Gly Phe Ala Phe Pro Arg Leu Pro Ala Pro Leu 
                645                 650                 655     


His Leu Ala Asp Arg Leu Val Lys Glu Val Gly Arg Leu Gly Ile Arg 
            660                 665                 670         


His Leu Lys Glu Val Asp Arg Glu Lys Leu Phe Phe Val Gly Ser Glu 
        675                 680                 685             


Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly 
    690                 695                 700                 


Pro Ser Gly Ser Glu Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr 
705                 710                 715                 720 


Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr Ser Glu Gly 
                725                 730                 735     


Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val 
            740                 745                 750         


Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe 
        755                 760                 765             


Leu Tyr Gly Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp 
    770                 775                 780                 


Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr 
785                 790                 795                 800 


Thr Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu 
                805                 810                 815     


Gln Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe 
            820                 825                 830         


Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala 
        835                 840                 845             


Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn 
    850                 855                 860                 


Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile 
865                 870                 875                 880 


Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro 
                885                 890                 895     


Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn 
            900                 905                 910         


Asn Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys 
        915                 920                 925             


Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn 
    930                 935                 




