
                                SEQUENCE LISTING

<110> Pioneer Hi-Bred Int'l., Inc.

<120> Methods for Altering the Genome of a
  Monocot Plant Cell

<130> 2332-PCT

<150> 60/947,003          
<151> 2007-06-29  

<150> 61/033,150          
<151> 2008-03-03  

<160> 23

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 720
<212> DNA
<213> Artificial Sequence

<220> 
<223> Maize codon-modified I-SceI coding sequence
      (moI-SceI)
      

<400> 1
atggccaaga acatcaagaa gaaccaggtg atgaacctgg gcccgaacag caagctcctg 60
aaggagtaca agtcgcagct catcgagctg aacatcgagc agttcgaggc cggcatcggc 120
ctcatcctgg gcgacgccta catcaggtcc cgcgacgagg gcaagaccta ctgcatgcag 180
ttcgagtgga agaacaaggc gtacatggac cacgtgtgcc tcctgtacga ccagtgggtc 240
ctcagcccgc cgcacaagaa ggagcgcgtg aaccacctgg gcaacctcgt catcacgtgg 300
ggcgcccaga ccttcaagca ccaggcgttc aacaagctgg ccaacctctt catcgtgaac 360
aacaagaaga cgatcccgaa caacctggtc gagaactacc tcacgcccat gtcgctggcg 420
tactggttca tggacgacgg cggcaagtgg gactacaaca agaactccac gaacaagagc 480
atcgtgctca acacccagtc gttcacgttc gaggaggtcg agtacctggt gaagggcctc 540
cggaacaagt tccagctcaa ctgctacgtc aagatcaaca agaacaagcc gatcatctac 600
atcgactcca tgagctacct catcttctac aacctgatca agccctacct catcccgcag 660
atgatgtaca agctgcccaa caccatctcg tccgagacgt tcctcaaggc cgcggactga 720


<210> 2
<211> 18
<212> DNA
<213> Saccharomyces cerevisiae

<220> 
<221> misc_feature   
<222> (1)...(18)
<223> I-SceI recognition sequence
      

<400> 2
tagggataac agggtaat                                               18

<210> 3
<211> 25
<212> DNA
<213> Artificial Sequence

<220> 
<223> ATS primer 104797

<400> 3
tcgtgaacca ctacatcgcg acctc                                       25

<210> 4
<211> 23
<212> DNA
<213> Artificial Sequence

<220> 
<223> ATS primer 104798

<400> 4
agcagcttgt gctggatgaa gtg                                         23

<210> 5
<211> 20
<212> DNA
<213> Artificial Sequence

<220> 
<223> I-SceI primer 59723

<400> 5
tcttgctggg cacgctcttg                                             20

<210> 6
<211> 25
<212> DNA
<213> Artificial Sequence

<220> 
<223> I-Sce1 primer 104381

<400> 6
ggtctcgatc tttggccttg gtagt                                       25

<210> 7
<211> 33
<212> DNA
<213> Artificial Sequence

<220> 
<223> HR primer 124641

<400> 7
taagattaaa atagctttcc cccgttgcag cgc                              33

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

<220> 
<223> HR primer 104798

<400> 8
agcagcttgt gctggatgaa gtg                                         23

<210> 9
<211> 26
<212> DNA
<213> Artificial Sequence

<220> 
<223> LB primer 104801

<400> 9
ttcgaagatc tgcccactag tgagtc                                      26

<210> 10
<211> 23
<212> DNA
<213> Artificial Sequence

<220> 
<223> LB primer 104798

<400> 10
agcagcttgt gctggatgaa gtg                                         23

<210> 11
<211> 25
<212> DNA
<213> Artificial Sequence

<220> 
<223> LB primer 104381

<400> 11
ggtctcgatc tttggccttg gtagt                                       25

<210> 12
<211> 30
<212> DNA
<213> Artificial Sequence

<220> 
<223> LB primer 124437

<400> 12
cttgttgaac gcctggtgct tgaaggtctg                                  30

<210> 13
<211> 30
<212> DNA
<213> Artificial Sequence

<220> 
<223> RB primer 124437

<400> 13
cttgttgaac gcctggtgct tgaaggtctg                                  30

<210> 14
<211> 23
<212> DNA
<213> Artificial Sequence

<220> 
<223> RB primer 104798

<400> 14
agcagcttgt gctggatgaa gtg                                         23

<210> 15
<211> 200
<212> DNA
<213> Zea mays

<220> 
<221> misc_feature   
<222> (0)...(0)
<223> LIG3-4 genomic target locus

<400> 15
cttcttttga tcggctgcgg aaataatata ctgtaacgat ttacgcacct gctgggaatt 60
gtaccgtacg tgccccggcg gaggatatat atacctcaca cgtacgcgta cgcgtatata 120
tacgtgcgct gctactcatt tgcgcgggaa tacagctcag tctgctgtgc gctgcaggat 180
gtacatacat acatgcgcag                                             200

<210> 16
<211> 22
<212> DNA
<213> Zea mays

<220> 
<221> misc_feature   
<222> (0)...(0)
<223> LIG3-4 recognition sequence

<400> 16
atatacctca cacgtacgcg ta                                          22

<210> 17
<211> 22
<212> DNA
<213> Chlamydomonas reinhardtii

<220> 
<221> misc_feature   
<222> (0)...(0)
<223> I-CreI recognition sequence

<400> 17
caaaacgtcg tgagacagtt tg                                          22

<210> 18
<211> 29
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer Lig34f Biocode 136247

<400> 18
taattaggga gagaaaaata gagcaccag                                   29

<210> 19
<211> 27
<212> DNA
<213> Artificial Sequence

<220> 
<223> Primer Lig34r Biocode 136248

<400> 19
atgtgcattg catcgctctt ctctctc                                     27

<210> 20
<211> 163
<212> PRT
<213> Chlamydomonas reinhardtii

<220> 
<223> I-CreI monomer
      

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


<210> 21
<211> 350
<212> PRT
<213> Artificial Sequence

<220> 
<223> LIG3-4 single chain custom meganuclease

<400> 21
Met Asn Thr Lys Tyr Asn Lys Glu Phe Leu Leu Tyr Leu Ala Gly Phe
 1               5                  10                  15      
Val Asp Gly Asp Gly Ser Ile Lys Ala Gln Ile Lys Pro Asn Gln Ser
            20                  25                  30          
Cys Lys Phe Lys His Gln Leu Ser Leu Thr Phe Gln Val Thr Gln Lys
        35                  40                  45              
Thr Gln Arg Arg Trp Phe Leu Asp Lys Leu Val Asp Glu Ile Gly Val
    50                  55                  60                  
Gly Tyr Val Tyr Asp Arg Gly Ser Val Ser Asp Tyr Glu Leu Ser Gln
65                  70                  75                  80  
Ile Lys Pro Leu His Asn Phe Leu Thr Gln Leu Gln Pro Phe Leu Lys
                85                  90                  95      
Leu Lys Gln Lys Gln Ala Asn Leu Val Leu Lys Ile Ile Glu Gln Leu
            100                 105                 110         
Pro Ser Ala Lys Glu Ser Pro Asp Lys Phe Leu Glu Val Cys Thr Trp
        115                 120                 125             
Val Asp Gln Ile Ala Ala Leu Asn Asp Ser Lys Thr Arg Lys Thr Thr
    130                 135                 140                 
Ser Glu Thr Val Arg Ala Val Leu Asp Ser Leu Pro Gly Ser Val Gly
145                 150                 155                 160 
Gly Leu Ser Pro Ser Gln Ala Ser Ser Ala Ala Ser Ser Ala Ser Ser
                165                 170                 175     
Ser Pro Gly Ser Gly Ile Ser Glu Ala Leu Arg Ala Gly Ala Thr Lys
            180                 185                 190         
Ser Lys Glu Phe Leu Leu Tyr Leu Ala Gly Phe Val Asp Gly Asp Gly
        195                 200                 205             
Ser Ile Ile Ala Ser Ile Lys Pro Arg Gln Cys Tyr Lys Phe Lys His
    210                 215                 220                 
Glu Leu Arg Leu Glu Phe Thr Val Thr Gln Lys Thr Gln Arg Arg Trp
225                 230                 235                 240 
Phe Leu Asp Lys Leu Val Asp Glu Ile Gly Val Gly Tyr Val Tyr Asp
                245                 250                 255     
Arg Gly Ser Val Ser Asp Tyr Arg Leu Ser Gln Ile Lys Pro Leu His
            260                 265                 270         
Asn Phe Leu Thr Gln Leu Gln Pro Phe Leu Lys Leu Lys Gln Lys Gln
        275                 280                 285             
Ala Asn Leu Val Leu Lys Ile Ile Glu Gln Leu Pro Ser Ala Lys Glu
    290                 295                 300                 
Ser Pro Asp Lys Phe Leu Glu Val Cys Thr Trp Val Asp Gln Ile Ala
305                 310                 315                 320 
Ala Leu Asn Asp Ser Lys Thr Arg Lys Thr Thr Ser Glu Thr Val Arg
                325                 330                 335     
Ala Val Leu Asp Ser Leu Ser Glu Lys Lys Lys Ser Ser Pro
            340                 345                 350 


<210> 22
<211> 163
<212> PRT
<213> Artificial Sequence

<220> 
<223> LIG3K nuclease monomer derived by modification of
      I-CreI endonuclease

<400> 22
Met Asn Thr Lys Tyr Asn Lys Lys Phe Leu Leu Tyr Leu Ala Gly Phe
 1               5                  10                  15      
Val Asp Gly Asp Gly Ser Ile Lys Ala Gln Ile Lys Pro Asn Gln Ser
            20                  25                  30          
Cys Lys Phe Lys His Gln Leu Ser Leu Thr Phe Gln Val Thr Gln Lys
        35                  40                  45              
Thr Gln Arg Arg Trp Phe Leu Asp Lys Leu Val Asp Lys Ile Gly Val
    50                  55                  60                  
Gly Tyr Val Tyr Asp Arg Gly Ser Val Ser Asp Tyr Glu Leu Ser Gln
65                  70                  75                  80  
Ile Lys Pro Leu His Asn Phe Leu Thr Gln Leu Gln Pro Phe Leu Lys
                85                  90                  95      
Leu Lys Gln Lys Gln Ala Asn Leu Val Leu Lys Ile Ile Glu Gln Leu
            100                 105                 110         
Pro Ser Ala Lys Glu Ser Pro Asp Lys Phe Leu Glu Val Cys Thr Trp
        115                 120                 125             
Val Asp Gln Ile Ala Ala Leu Asn Asp Ser Lys Thr Arg Lys Thr Thr
    130                 135                 140                 
Ser Glu Thr Val Arg Ala Val Leu Asp Ser Leu Ser Glu Lys Lys Lys
145                 150                 155                 160 
Ser Ser Pro
            


<210> 23
<211> 163
<212> PRT
<213> Artificial Sequence

<220> 
<223> LIG4E nuclease monomer derived by modification of
      I-CreI endonuclease

<400> 23
Met Asn Thr Lys Tyr Asn Glu Glu Phe Leu Leu Tyr Leu Ala Gly Phe
 1               5                  10                  15      
Val Asp Gly Asp Gly Ser Ile Ile Ala Ser Ile Lys Pro Arg Gln Cys
            20                  25                  30          
Tyr Lys Phe Lys His Glu Leu Arg Leu Glu Phe Thr Val Thr Gln Lys
        35                  40                  45              
Thr Gln Arg Arg Trp Phe Leu Asp Glu Leu Val Asp Glu Ile Gly Val
    50                  55                  60                  
Gly Tyr Val Arg Asp Arg Gly Ser Val Ser Asp Tyr Arg Leu Ser Gln
65                  70                  75                  80  
Ile Lys Pro Leu His Asn Phe Leu Thr Gln Leu Gln Pro Phe Leu Glu
                85                  90                  95      
Leu Lys Gln Lys Gln Ala Asn Leu Val Leu Lys Ile Ile Glu Gln Leu
            100                 105                 110         
Pro Ser Ala Lys Glu Ser Pro Asp Lys Phe Leu Glu Val Cys Thr Trp
        115                 120                 125             
Val Asp Gln Ile Ala Ala Leu Asn Asp Ser Lys Thr Arg Lys Thr Thr
    130                 135                 140                 
Ser Glu Thr Val Arg Ala Val Leu Asp Ser Leu Ser Glu Lys Lys Lys
145                 150                 155                 160 
Ser Ser Pro
            

