                         SEQUENCE LISTING

<110>  North Carolina State University
       Barrangou, Rodolphe
       Briner, Alexandra E.
 
<120>  COMPOSITIONS AND METHODS RELATED TO A TYPE-II CRISPR-CAS SYSTEM 
       IN LACTOBACILLUS BUCHNERI

<130>  5051-844WO

<150>  US 61/897,670
<151>  2013-10-30

<160>  63    

<170>  PatentIn version 3.5

<210>  1
<211>  1371
<212>  PRT
<213>  Lactobacillus buchneri

<400>  1

Met Lys Val Asn Asn Tyr His Ile Gly Leu Asp Ile Gly Thr Ser Ser 
1               5                   10                  15      


Ile Gly Trp Val Ala Ile Gly Glu Asp Gly Lys Pro Leu Arg Ile Lys 
            20                  25                  30          


Gly Lys Thr Ala Ile Gly Ala Arg Leu Phe Gln Glu Gly Asn Pro Ala 
        35                  40                  45              


Ala Asp Arg Arg Met Phe Arg Thr Thr Arg Arg Arg Leu Ser Arg Arg 
    50                  55                  60                  


Lys Trp Arg Leu Lys Leu Leu Glu Glu Ile Phe Asp Pro Tyr Ile Thr 
65                  70                  75                  80  


Pro Val Asp Ser Thr Phe Phe Ala Arg Leu Lys Gln Ser Asn Leu Ser 
                85                  90                  95      


Pro Lys Asp Ser Arg Lys Glu Phe Lys Gly Ser Met Leu Phe Pro Asp 
            100                 105                 110         


Leu Thr Asp Met Gln Tyr His Lys Asp Tyr Pro Thr Ile Tyr His Leu 
        115                 120                 125             


Arg His Ala Leu Met Thr Gln Asp Glu Lys Phe Asp Ile Arg Met Val 
    130                 135                 140                 


Tyr Leu Ala Ile His His Ile Val Lys Tyr Arg Gly Asn Phe Leu Asn 
145                 150                 155                 160 


Ser Thr Pro Val Asp Ser Phe Lys Ala Ser Lys Val Asn Phe Val Asp 
                165                 170                 175     


Gln Phe Lys Lys Leu Asn Glu Leu Tyr Thr Ala Ile Asn Pro Glu Glu 
            180                 185                 190         


Ser Phe Gln Ile Asn Leu Ala Asn Ser Glu Asp Ile Gly His Gln Phe 
        195                 200                 205             


Leu Asp Pro Ser Ile Arg Lys Phe Asp Lys Lys Lys Gln Ile Pro Lys 
    210                 215                 220                 


Ile Val Pro Val Ser Val Asp Asp Lys Ala Thr Asp Lys Ile Asn Gly 
225                 230                 235                 240 


Lys Ile Ala Ser Glu Ile Ile Asn Ala Ile Leu Gly Tyr Lys Ser Lys 
                245                 250                 255     


Leu Asp Val Val Val Gln Cys Thr Pro Val Asp Ser Lys Ser Trp Ala 
            260                 265                 270         


Leu Lys Phe Asp Glu Glu Asp Ile Asp Ala Lys Leu Gln Lys Ile Leu 
        275                 280                 285             


Pro Glu Met Asp Glu Asn Gln Gln Ser Ile Ile Ala Ile Leu Gln Asn 
    290                 295                 300                 


Leu Tyr Ser Gln Val Thr Leu Asn Gln Ile Val Pro Asn Gly Met Ser 
305                 310                 315                 320 


Leu Ser Glu Ser Met Ile Glu Lys Tyr Asn Asp His His Asp His Leu 
                325                 330                 335     


Lys Leu Tyr Lys Lys Ile Ile Asp Gln Leu Ala Asp Pro Lys Lys Lys 
            340                 345                 350         


Ala Ala Leu Lys Lys Ala Tyr Ser Gln Tyr Val Gly Asp Asp Gly Lys 
        355                 360                 365             


Val Ile Glu Gln Ala Asp Phe Trp Ser Ser Val Lys Lys Asn Leu Asp 
    370                 375                 380                 


Asp Ser Asp Leu Ser Lys Gln Ile Met Asp Leu Ile Asp Ala Glu Lys 
385                 390                 395                 400 


Phe Met Pro Lys Gln Arg Thr Ser Gln Asn Gly Val Ile Pro His Gln 
                405                 410                 415     


Leu His Gln Arg Glu Leu Asp Glu Ile Ile Glu His Gln Ser Lys Tyr 
            420                 425                 430         


Tyr Pro Trp Leu Ala Glu Ile Asn Pro Asn Lys His Asp Leu His Leu 
        435                 440                 445             


Ala Lys Tyr Lys Ile Glu Glu Leu Val Ala Phe Arg Val Pro Tyr Tyr 
    450                 455                 460                 


Val Gly Pro Met Ile Thr Pro Asp Asp Gln Ala Lys Ser Ala Glu Thr 
465                 470                 475                 480 


Val Phe Ser Trp Met Glu Arg Lys Gly Lys Glu Ala Gly Gln Ile Thr 
                485                 490                 495     


Pro Trp Asn Phe Asp Glu Lys Val Asp Arg Asn Ala Ser Ala Asn Arg 
            500                 505                 510         


Phe Ile Lys Arg Met Thr Thr Lys Asp Thr Tyr Leu Ile Gly Glu Asp 
        515                 520                 525             


Val Leu Pro Asp Glu Ser Leu Leu Tyr Glu Lys Phe Lys Val Leu Asn 
    530                 535                 540                 


Glu Leu Asn Met Val Arg Val Asn Gly Lys Leu Leu Lys Val Ala Asp 
545                 550                 555                 560 


Lys Gln Ala Ile Phe Gln Asp Leu Phe Glu Asn Tyr Lys His Ile Ser 
                565                 570                 575     


Val Lys Lys Leu Gln Asn Tyr Ile Lys Ser Lys Thr Gly Leu Pro Ser 
            580                 585                 590         


Asp Pro Glu Ile Ser Gly Leu Ser Asp Pro Glu Tyr Phe Asn Asn Ser 
        595                 600                 605             


Leu Gly Thr Tyr Asn Asp Phe Lys Lys Leu Phe Gly Asn Lys Val Asp 
    610                 615                 620                 


Glu Pro Asp Leu Gln Asp Asp Phe Glu Lys Ile Val Glu Trp Ser Thr 
625                 630                 635                 640 


Val Phe Glu Asp Lys Arg Ile Leu Arg Glu Lys Leu Asn Glu Ile Thr 
                645                 650                 655     


Trp Leu Ser Asp Gln Gln Lys Asp Val Leu Glu Ser Ser Arg Tyr Gln 
            660                 665                 670         


Gly Trp Gly Arg Leu Ser Lys Lys Leu Leu Thr Gly Ile Val Asn Asp 
        675                 680                 685             


Gln Gly Glu Arg Ile Ile Asp Glu Leu Trp Asn Thr Asn Lys Asn Phe 
    690                 695                 700                 


Met Gln Ile Gln Ser Asp Asn Asp Phe Ala Lys Arg Ile His Glu Ala 
705                 710                 715                 720 


Asn Ala Asp Gln Met Lys Ala Val Asp Val Glu Asp Val Leu Ala Asp 
                725                 730                 735     


Ala Tyr Thr Ser Pro Gln Asn Lys Lys Ala Ile Arg Gln Val Val Lys 
            740                 745                 750         


Val Val Asp Asp Ile Gln Lys Ala Met Gly Gly Val Ala Pro Lys Tyr 
        755                 760                 765             


Ile Ser Ile Glu Phe Thr Arg Ser Glu Asp Arg Asn Pro Arg Arg Thr 
    770                 775                 780                 


Ile Ser Arg Gln Arg Gln Leu Glu Asn Thr Leu Lys Asp Thr Ala Lys 
785                 790                 795                 800 


Ser Leu Ala Lys Ser Ile Asn Pro Glu Leu Leu Ser Glu Leu Asp Asn 
                805                 810                 815     


Ala Ala Lys Ser Lys Lys Gly Leu Thr Asp Arg Leu Tyr Leu Tyr Phe 
            820                 825                 830         


Thr Gln Leu Gly Lys Asp Ile Tyr Thr Gly Lys Pro Ile Asn Ile Asp 
        835                 840                 845             


Glu Ile Ser Thr Tyr Asp Ile Asp His Ile Leu Pro Gln Ala Phe Val 
    850                 855                 860                 


Lys Asp Asp Ser Leu Asn Asn Arg Val Leu Val Ser Lys Ala Ile Asn 
865                 870                 875                 880 


Asn Gly Lys Ser Asp Asn Val Pro Val Gln Leu Phe Gly Ala Lys Met 
                885                 890                 895     


Gly His Phe Trp Lys Gln Leu Ala Glu Ala Gly Leu Ile Ser Lys Arg 
            900                 905                 910         


Lys Leu Lys Asn Leu Gln Thr Asp Pro Asp Thr Ile Ser Lys Tyr Ala 
        915                 920                 925             


Met His Gly Phe Ile Arg Arg Gln Leu Val Glu Thr Ser Gln Val Ile 
    930                 935                 940                 


Lys Leu Val Ala Asn Ile Leu Gly Asp Lys Tyr Arg Asn Asp Asn Thr 
945                 950                 955                 960 


Lys Ile Ile Glu Ile Thr Ala Arg Met Asn His Gln Met Arg Asp Glu 
                965                 970                 975     


Phe Gly Phe Ile Lys Asn Arg Glu Ile Asn Asp Tyr His His Ala Phe 
            980                 985                 990         


Asp Ala Tyr Leu Thr Ala Phe Leu  Gly Arg Tyr Leu Tyr  His Arg Tyr 
        995                 1000                 1005             


Ile Lys  Leu Arg Pro Tyr Phe  Val Tyr Gly Asp Phe  Lys Lys Phe 
    1010                 1015                 1020             


Lys Glu  Asp Lys Val Thr Met  Arg Asn Phe Asn Phe  Leu His Asp 
    1025                 1030                 1035             


Leu Thr  Asp Asp Thr Gln Glu  Lys Ile Ala Asp Ala  Glu Thr Gly 
    1040                 1045                 1050             


Glu Val  Ile Trp Asp Arg Glu  Asn Ser Ile Gln Gln  Leu Lys Asp 
    1055                 1060                 1065             


Val Tyr  His Tyr Lys Phe Met  Leu Ile Ser His Glu  Val Tyr Thr 
    1070                 1075                 1080             


Leu Arg  Gly Ala Met Phe Asn  Gln Thr Val Tyr Pro  Ala Ser Asp 
    1085                 1090                 1095             


Ala Gly  Lys Arg Lys Leu Ile  Pro Ile Lys Ala Asp  Arg Pro Ile 
    1100                 1105                 1110             


Asn Val  Tyr Gly Gly Tyr Ser  Gly Ser Ala Asp Ala  Tyr Met Ala 
    1115                 1120                 1125             


Ile Val  Arg Ile His Asn Lys  Lys Gly Asp Lys Tyr  Arg Val Val 
    1130                 1135                 1140             


Gly Val  Pro Met Arg Ala Arg  Asp Arg Leu Asp Ala  Ala Lys Lys 
    1145                 1150                 1155             


Val Ser  Asp Ala Asp Cys Asp  Arg Ala Leu Lys Asp  Val Leu Thr 
    1160                 1165                 1170             


Pro Gln  Leu Thr Lys Thr Lys  Lys Ser Arg Lys Thr  Gly Glu Ile 
    1175                 1180                 1185             


Thr Gln  Val Val Glu Asp Phe  Glu Ile Val Leu Gly  Lys Val Met 
    1190                 1195                 1200             


Tyr Arg  Gln Leu Met Ile Asp  Gly Asp Lys Lys Phe  Met Leu Gly 
    1205                 1210                 1215             


Ser Ser  Thr Tyr Gln Tyr Asn  Ala Lys Gln Leu Val  Leu Ser Asp 
    1220                 1225                 1230             


Gln Ser  Val Lys Thr Leu Ala  Ser Lys Gly Arg Leu  Asp Pro Leu 
    1235                 1240                 1245             


Gln Glu  Ser Met Asp Tyr Asn  Asn Val Tyr Thr Glu  Ile Leu Asp 
    1250                 1255                 1260             


Lys Val  Asn Gln Tyr Phe Ser  Leu Tyr Asp Met Asn  Lys Phe Arg 
    1265                 1270                 1275             


His Lys  Leu Asn Leu Gly Phe  Ser Lys Phe Ile Ser  Phe Pro Asn 
    1280                 1285                 1290             


His Asn  Val Phe Asp Gly Asn  Thr Lys Ala Ser Ser  Gly Lys Arg 
    1295                 1300                 1305             


Glu Ile  Leu Glu Glu Val Leu  Asn Gly Leu His Ala  Asn Pro Thr 
    1310                 1315                 1320             


Phe Gly  Asn Leu Lys Asp Ile  Gly Ile Thr Thr Pro  Phe Gly Gln 
    1325                 1330                 1335             


Leu Gln  Gln Pro Asn Gly Ile  Leu Leu Ser Asp Glu  Ala Lys Ile 
    1340                 1345                 1350             


Arg Tyr  Gln Ser Pro Thr Gly  Leu Phe Glu Arg Thr  Val Ser Leu 
    1355                 1360                 1365             


Lys Asp  Leu 
    1370     


<210>  2
<211>  36
<212>  DNA
<213>  Lactobacillus buchneri

<400>  2
gttttagaag gatgttaaat caataaggtt aaaccc                                 36


<210>  3
<211>  20
<212>  DNA
<213>  Lactobacillus buchneri

<400>  3
auuuaacauc cuguguuaaa                                                   20


<210>  4
<211>  18
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  4
gctttagtag ttcaaaac                                                     18


<210>  5
<211>  22
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  5
catcattgtt ttgaactact ac                                                22


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

<220>
<223>  PCR primer

<400>  6
gggtttaacc ttattgattt aac                                               23


<210>  7
<211>  22
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  7
gaaggatgtt aaatcaataa gg                                                22


<210>  8
<211>  18
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  8
ccttcagact gacggttc                                                     18


<210>  9
<211>  19
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  9
gtctcgatat tgggacctc                                                    19


<210>  10
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  10
ccagaatgaa tgatctgttg                                                   20


<210>  11
<211>  17
<212>  DNA
<213>  Artificial

<220>
<223>  PCR primer

<400>  11
catcgacgag aactttg                                                      17


<210>  12
<211>  7
<212>  PRT
<213>  Lactobacillus buchneri

<400>  12

Ile Gly Leu Asp Ile Gly Thr 
1               5           


<210>  13
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  L. buchneri conserved sequence


<220>
<221>  misc_feature
<222>  (2)..(2)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  misc_feature
<222>  (4)..(4)
<223>  Xaa can be any naturally occurring amino acid

<400>  13

His Xaa Asn Xaa Asn 
1               5   


<210>  14
<211>  6
<212>  PRT
<213>  Lactobacillus buchneri

<400>  14

Tyr Asp Ile Asp His Ile 
1               5       


<210>  15
<211>  5
<212>  PRT
<213>  Lactobacillus buchneri

<400>  15

Asn Asn Arg Val Leu 
1               5   


<210>  16
<211>  4
<212>  PRT
<213>  Lactobacillus buchneri

<400>  16

Ile Asn Asn Gly 
1               


<210>  17
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  17
aaaattcaga                                                              10


<210>  18
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  18
caacaaaaaa agcgctccgc aacggccatt                                        30


<210>  19
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  19
gcaaaacgct                                                              10


<210>  20
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  20
atgaagttca                                                              10


<210>  21
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  21
agctgtgtca aactacgttg aatcccaagg                                        30


<210>  22
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  22
acaaaactta                                                              10


<210>  23
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  23
ctggttttat                                                              10


<210>  24
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  24
aaacggatat tgcggcttat attaacgagc                                        30


<210>  25
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  25
tgaaatggtt                                                              10


<210>  26
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  26
ctggttttat                                                              10


<210>  27
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  27
aaacggatat tgcggcttat attaacgagc                                        30


<210>  28
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  28
tgaaatggtt                                                              10


<210>  29
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  29
agaatatcga                                                              10


<210>  30
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  30
caacgcagct aaagataatc gtcagaatta                                        30


<210>  31
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  31
ccagaaatta                                                              10


<210>  32
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  32
taagcttggt                                                              10


<210>  33
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  33
ggaaaaaggt ggcggccgct ttgtgcaagg                                        30


<210>  34
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  34
tcaagaaatg                                                              10


<210>  35
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  35
ttacgcttta                                                              10


<210>  36
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  36
accgagtttc gtgatctcaa aagtagctac                                        30


<210>  37
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  37
gcaaaaacta                                                              10


<210>  38
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  38
ttcttagatg                                                              10


<210>  39
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  39
ccgcttactt gccgttaaag cgggatatcg                                        30


<210>  40
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  40
ttcaaaaaga                                                              10


<210>  41
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  41
aaaattcaga                                                              10


<210>  42
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  42
caacaaaaaa agcgctccgc aacggccatt                                        30


<210>  43
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  43
gcaaaacgct                                                              10


<210>  44
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  44
cattatgcta                                                              10


<210>  45
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  45
aaggttcagg tgtctcacac gctgaactag                                        30


<210>  46
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  46
acaaaattat                                                              10


<210>  47
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  47
ctttatctag                                                              10


<210>  48
<211>  30
<212>  DNA
<213>  Lactobacillus buchneri

<400>  48
gaaataagca gcctcatttg aagcaccatg                                        30


<210>  49
<211>  10
<212>  DNA
<213>  Lactobacillus buchneri

<400>  49
ccaaaaatga                                                              10


<210>  50
<211>  28
<212>  DNA
<213>  Lactobacillus buchneri

<400>  50
gtattcccca cgtacgtagg ggtgatcc                                          28


<210>  51
<211>  28
<212>  DNA
<213>  Lactobacillus buchneri

<400>  51
gtattcccca cgtgtgtagg ggtgatcc                                          28


<210>  52
<211>  28
<212>  DNA
<213>  Lactobacillus buchneri

<400>  52
gtattcccca tgtatgtggg ggtgatcc                                          28


<210>  53
<211>  28
<212>  DNA
<213>  Lactobacillus brevis

<400>  53
gtattcccca cacatgtggg ggtgatcc                                          28


<210>  54
<211>  28
<212>  DNA
<213>  Lactobacillus brevis

<400>  54
gtattcccca caggtgtggg ggtgatcc                                          28


<210>  55
<211>  50
<212>  DNA
<213>  Lactobacillus buchneri

<400>  55
cattatgcta aaggttcagg tgtctcacac gctgaactca acaaaattat                  50


<210>  56
<211>  102
<212>  DNA
<213>  Lactobacillus buchneri

<400>  56
gttttagaag gatgttaaat caataaggtt aaacccaagg ttcaggtgtc tcacacgctg       60

aactcagttt tagaaggatg ttaaatcaat aaggttaaac cc                         102


<210>  57
<211>  90
<212>  DNA
<213>  Lactobacillus buchneri

<400>  57
auguuugacc uuauugauuu aacauccugu guuaaaauca agcaaagcgc uuugcgcgga       60

guuucaacuu uugacccauu auaugggcau                                        90


<210>  58
<211>  40
<212>  DNA
<213>  Lactobacillus buchneri

<400>  58
ugucucacac gcugaacuca guuuuagaag gauguuaaau                             40


<210>  59
<211>  50
<212>  DNA
<213>  Lactobacillus buchneri

<400>  59
ataattttgt tgagttcagc gtgtgagaca cctgaacctt tagcataatg                  50


<210>  60
<211>  201
<212>  DNA
<213>  Lactobacillus buchneri

<400>  60
gaggtgactg tcggagagag ggaggatgca guuuuagaag gauguuaaau caauaagguu       60

aaacguuuga ccuuauugau uuaacauccu guguugaaau caagcaaagc gcuuugcgcg      120

gaguuucaac uuuugaccca uuauaugggc auuacauaag cgaaaggaac cauucuucag      180

uuggagaaug guuccuuuuu u                                                201


<210>  61
<211>  36
<212>  DNA
<213>  Lactobacillus buchneri


<220>
<221>  misc_feature
<222>  (5)..(6)
<223>  n is a, c, g, t or u

<400>  61
ttttnntgta tcctcactct ctccgacagt cacctc                                 36


<210>  62
<211>  35
<212>  DNA
<213>  Lactobacillus buchneri


<220>
<221>  misc_feature
<222>  (31)..(32)
<223>  n is a, c, g, t or u

<400>  62
gaggtgactg tcggagagag tgaggataca nnaaa                                  35


<210>  63
<211>  185
<212>  DNA
<213>  Lactobacillus buchneri

<400>  63
gaggtgactg tcggagagag ggaggatgca guuuuagaag gauguuaaau caauuguuau       60

ugauuuaaca uccuguguug aaaucaagca aagcgcuuug cgcggaguuu caacuuuuga      120

cccauuauau gggcauuaca uaagcgaaag gaaccauucu ucaguuggag aaugguuccu      180

uuuuu                                                                  185


