                         SEQUENCE LISTING

<110>  Zhong, Minghong
 
<120>  Conjugates of Guide RNA-Cas Protein Complex

<130>  01-00002

<150>  US62/888,551
<151>  2019-08-19

<150>  US62/914,565
<151>  2019-10-14

<150>  US62/937,876
<151>  2019-11-20

<160>  72    

<170>  PatentIn version 3.5

<210>  1
<211>  42
<212>  RNA
<213>  Streptococcus pyogenes


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  1
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug                          42


<210>  2
<211>  81
<212>  RNA
<213>  Streptococcus pyogenes

<400>  2
ggaaccauuc aaaacagcau agcaaguuaa aauaaggcua guccguuauc aacuugaaaa       60

aguggcaccg agucggugcu u                                                 81


<210>  3
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA, nn (32, 33) is either base-paired  or not, and joined by 
       non-nucleotide linker between the two, n32 (A) and n33 (B).


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, or u

<400>  3
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  4
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  sgRNA: crRNA and tracrRNA are fused as a single whole RNA by a 
       tetraloop, gaaa.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  4
nnnnnnnnnn nnnnnnnnnn guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  5
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (32, 33) of crgRNA and 
       tracrgRNA, and between a70 and a71 in tracrgRNA.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, or u

<400>  5
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  6
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  gRNA inserted with an aptamer between a32 and u33.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  6
nnnnnnnnnn nnnnnnnnnn guuuuagagc uauagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  7
<211>  92
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  gRNA inserted with two aptamers, positioned between a32 and u33 
       and between g69 and a70.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  7
nnnnnnnnnn nnnnnnnnnn guuuuagagc uauagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaguggcacc gagucggugc uu                                     92


<210>  8
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA inserted with two chemically ligated aptamers by 
       nNt-linkers, positioned between n32 and n33 and between a70 and 
       a71.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, or u

<400>  8
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  9
<211>  62
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Chemically ligated tracrRNA inserted with two aptamers by 
       nNt-linkers, positioned at 5'-end  and n33 and between a38 and 
       a39.


<220>
<221>  misc_feature
<222>  (1)..(1)
<223>  n is a, c, g, or u

<400>  9
nagcaaguua aaauaaggcu aguccguuau caacuugaaa aaguggcacc gagucggugc       60

uu                                                                      62


<210>  10
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA inserted with two chemically ligated aptamers at 
       nNt-linkers, positioned between n32 and n33 and between a70 and 
       a71.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, or u

<400>  10
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  11
<211>  36
<212>  RNA
<213>  Streptococcus pyogenes


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  11
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcu                                 36


<210>  12
<211>  66
<212>  RNA
<213>  Streptococcus pyogenes

<400>  12
agcauagcaa guuaaaauaa ggcuaguccg uuaucaacuu gaaaaagugg caccgagucg       60

gugcuu                                                                  66


<210>  13
<211>  47
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA (partial sequence) formed by chemical ligation bwteen 
       tracrRNA and crRNA. The nNt-linker is positioned bwteen c15 
       (3'-end of tracrRNA) and g16 (5'-end of crRNA).


<220>
<221>  misc_feature
<222>  (28)..(47)
<223>  n is a, c, g, or u

<400>  13
ggaagggaac ccuccggugg guuaaagnnn nnnnnnnnnn nnnnnnn                     47


<210>  14
<211>  94
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA, nn (32, 33) is either base-paired  or not, and joined by 
       non-nucleotide linker between the two, n32 (A) and n33 (B). GalAc
       ligands are conjugated at 3'-end of lgRNA via an nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, or u

<400>  14
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  15
<211>  47
<212>  RNA
<213>  Artificial Sequence

<220>
<223>   lgRNA (partial sequence) formed by chemical ligation bwteen 
       tracrRNA and crRNA. The nNt-linker of lgRNA is positioned bwteen 
       c15 (3'-end of tracrRNA) and g16 (5'-end of crRNA). GalAc ligands
       are conjugated at 3'-end of lgRNA via a second nNt-Linker.


<220>
<221>  misc_feature
<222>  (28)..(47)
<223>  n is a, c, g, or u

<400>  15
ggaagggaac ccuccggugg guuaaagnnn nnnnnnnnnn nnnnnnn                     47


<210>  16
<211>  42
<212>  RNA
<213>  nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  16
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug                          42


<210>  17
<211>  81
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  GalAc ligands are conjugated at 3'-end of tracrRNA via an 
       nNt-Linker.

<400>  17
ggaaccauuc aaaacagcau agcaaguuaa aauaaggcua guccguuauc aacuugaaaa       60

aguggcaccg agucggugcu u                                                 81


<210>  18
<211>  42
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  A cholesterol is conjugated at 3'-end of crRNA via an nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  18
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug                          42


<210>  19
<211>  42
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Tocopherol is conjugated at 3'-end of crRNA via an nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  19
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug                          42


<210>  20
<211>  42
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  An aptamer is conjugated at 3'-end of crRNA via an nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  20
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug                          42


<210>  21
<211>  42
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  An antibody is conjugated at 3'-end of crRNA via an nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  21
nnnnnnnnnn nnnnnnnnnn guuuuagagc uaugcuguuu ug                          42


<210>  22
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  General scheme of a target DNA to be edited. Its non-target 
       strand is removed.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (24)..(25)
<223>  n is a, c, g, or t

<400>  22
nnnnnnnnnn nnnnnnnnnn tggnn                                             25


<210>  23
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  A crRNA-ssDNA conjugate via an nNt-Linker, n1 to n2o is a spacer,
       n33 to n60 is an ssDNA, which is conjugated to 3-end of crRNA via
       an nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (32)..(60)
<223>  n is a, c, g, t or u

<400>  23
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnnnnnnnn nnnnnnnnnn nnnnnnnnnn       60


<210>  24
<211>  117
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  An ssDNA-tracrRNA conjugate via an nNt-Linker. n1 to n50 is an 
       ssDNA, which is conjugated to n51 (5'-end) of tracrRNA via an 
       nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(51)
<223>  n is a, c, g, t or u

<400>  24
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nagcauagca       60

aguuaaaaua aggcuagucc guuaucaacu ugaaaaagug gcaccgaguc ggugcuu         117


<210>  25
<211>  94
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (32, 33) of crRNA and 
       tracrRNA, and between a70 and a71 in tracrRNA. An ssDNA is 
       conjugated at the ligation site bwteen n32 and n33.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, t or u

<400>  25
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  26
<211>  94
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (32, 33) of crRNA and 
       tracrRNA, and between a70 and a71 in tracrRNA. An ssDNA is 
       conjugated at the ligation site bwteen n70 and n71.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, t or u

<400>  26
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuu                                   94


<210>  27
<211>  144
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (32, 33) of crRNA and 
       tracrRNA, and between a70 and a71 in tracrRNA. An ssDNA (n95 to 
       n144) is conjugated at the l3'-end of lgRNA via a nNt-Linker or 
       nucleotide linker.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (95)..(144)
<223>  n is a, c, g, t or u

<400>  27
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuunnnnnn nnnnnnnnnn nnnnnnnnnn      120

nnnnnnnnnn nnnnnnnnnn nnnn                                             144


<210>  28
<211>  111
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  tracrRNA formed by ligation between a37 and a38. An ssDNA (n62 to
       n111) is conjugated at the l3'-end of tracrRNA via a nNt-Linker 
       or nucleotide linker.


<220>
<221>  misc_feature
<222>  (62)..(111)
<223>  n is a, c, g, t or u

<400>  28
agcaaguuaa aauaaggcua guccguuauc aacuugaaaa aguggcaccg agucggugcu       60

unnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn n               111


<210>  29
<211>  144
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (82, 83) of crRNA and 
       tracrRNA, and between a120 and a121 in tracrRNA. An ssDNA (n1 to 
       n50) is conjugated at the 5'-end of lgRNA via a nNt-Linker or 
       nucleotide linker.


<220>
<221>  misc_feature
<222>  (1)..(70)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (82)..(83)
<223>  n is a, c, g, t or u

<400>  29
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn       60

nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu aucaacuuga      120

aaaaguggca ccgagucggu gcuu                                             144


<210>  30
<211>  144
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (82, 83) of crRNA and 
       tracrRNA, and between a120 and a121 in tracrRNA. An ssDNA (n1 to 
       n50) is conjugated at the 5'-end of lgRNA via a triazole 
       nNt-Linker.


<220>
<221>  misc_feature
<222>  (1)..(70)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (82)..(83)
<223>  n is a, c, g, t or u

<400>  30
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn       60

nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu aucaacuuga      120

aaaaguggca ccgagucggu gcuu                                             144


<210>  31
<211>  95
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between u33 and n34 of crRNA and 
       n35 of tracrRNA, and between a71 and a72 in tracrRNA. An ssDNA 
       (n1 to n50) is conjugated at the ligation site between nn (32, 
       33) via an nNt-Linker between the 3'-end of ssDNA and u33.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

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

<220>
<221>  misc_feature
<222>  (34)..(34)
<223>  n is a, c, g, t or u

<400>  31
nnnnnnnnnn nnnnnnnnnn guuuuagagc ununagcaag uuaaaauaag gcuaguccgu       60

uaucaacuug aaaaaguggc accgagucgg ugcuu                                  95


<210>  32
<211>  144
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between nn (32, 33) of crRNA and 
       tracrRNA, and between a70 and a71 in tracrRNA. An ssDNA (n95 to 
       n144) is conjugated at the l3'-end of lgRNA via a triazole 
       nNt-Linker between the 3'-end of lgRNA and the 5'-end of ssDNA.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (95)..(144)
<223>  n is a, c, g, t or u

<400>  32
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuunnnnnn nnnnnnnnnn nnnnnnnnnn      120

nnnnnnnnnn nnnnnnnnnn nnnn                                             144


<210>  33
<211>  95
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA formed by two ligations between u33 and n34 of crRNA and 
       n35 of tracrRNA, and between a71 and a72 in tracrRNA. An ssDNA 
       (n1 to n50) is conjugated at the ligation site between nn (32, 
       33) via an nNt-Linker between the 5'-end of ssDNA and u33.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

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

<220>
<221>  misc_feature
<222>  (34)..(34)
<223>  n is a, c, g, t or u

<400>  33
nnnnnnnnnn nnnnnnnnnn guuuuagagc ununagcaag uuaaaauaag gcuaguccgu       60

uaucaacuug aaaaaguggc accgagucgg ugcuu                                  95


<210>  34
<211>  194
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  An ssDNA (n145 to n194) is conjugated at the l3'-end of lgRNA via
       a triazole nNt-Linker between the 3'-end of lgRNA and the 5'-end 
       of ssDNA, and a second ssDNA s conjugated at the 5'-end of lgRNA 
       by thiol-maleimide coupling.


<220>
<221>  misc_feature
<222>  (1)..(70)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (82)..(83)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (145)..(194)
<223>  n is a, c, g, t or u

<400>  34
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn       60

nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu aucaacuuga      120

aaaaguggca ccgagucggu gcuunnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn      180

nnnnnnnnnn nnnn                                                        194


<210>  35
<211>  144
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  An ssDNA (n95 to n144) is conjugated at the l3'-end of lgRNA via 
       a triazole nNt-Linker between the 3'-end of lgRNA and a dsDNA. 
       Its complementary strand is removed.


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (32)..(33)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (95)..(144)
<223>  n is a, c, g, t or u

<400>  35
nnnnnnnnnn nnnnnnnnnn guuuuagagc unnagcaagu uaaaauaagg cuaguccguu       60

aucaacuuga aaaaguggca ccgagucggu gcuunnnnnn nnnnnnnnnn nnnnnnnnnn      120

nnnnnnnnnn nnnnnnnnnn nnnn                                             144


<210>  36
<211>  23
<212>  DNA
<213>  HBV

<400>  36
ctctgctaga tcccagagtg agg                                               23


<210>  37
<211>  23
<212>  DNA
<213>  HBV

<400>  37
gctatcgctg gatgtgtctg cgg                                               23


<210>  38
<211>  25
<212>  DNA
<213>  HBV

<400>  38
tggacttctc tcaattttct agggg                                             25


<210>  39
<211>  23
<212>  DNA
<213>  HBV

<400>  39
gggggatcac ccgtgtgtct tgg                                               23


<210>  40
<211>  25
<212>  DNA
<213>  HBV

<400>  40
tatgtggatg atgtggtact ggggg                                             25


<210>  41
<211>  21
<212>  DNA
<213>  HBV

<400>  41
cctcaccata cagcactcgg g                                                 21


<210>  42
<211>  26
<212>  DNA
<213>  HBV

<400>  42
gtgttggggt gagttgatga atctgg                                            26


<210>  43
<211>  25
<212>  DNA
<213>  HBV

<400>  43
tatgtggatg atgtggtact ggggg                                             25


<210>  44
<211>  25
<212>  DNA
<213>  HBV

<400>  44
tatatggatg atgtggtatt ggggg                                             25


<210>  45
<211>  25
<212>  DNA
<213>  HBV

<400>  45
tatgtggatg atgtggtatt ggggg                                             25


<210>  46
<211>  25
<212>  DNA
<213>  HBV

<400>  46
tatatagatg atgtggtact ggggg                                             25


<210>  47
<211>  23
<212>  DNA
<213>  HIV

<400>  47
gattggcaga actacacacc agg                                               23


<210>  48
<211>  23
<212>  DNA
<213>  HIV

<400>  48
atcagatatc cactgacctt tgg                                               23


<210>  49
<211>  23
<212>  DNA
<213>  HIV

<400>  49
gcgtggcctg ggcgggactg ggg                                               23


<210>  50
<211>  23
<212>  DNA
<213>  HIV

<400>  50
cagcagttct tgaagtactc cgg                                               23


<210>  51
<211>  23
<212>  DNA
<213>  EBV

<400>  51
gccctggacc aacccggccc ggg                                               23


<210>  52
<211>  23
<212>  DNA
<213>  EBV

<400>  52
ggccgctgcc ccgctccggg tgg                                               23


<210>  53
<211>  22
<212>  DNA
<213>  EBV

<400>  53
ggaagacaat gtgccgccat gg                                                22


<210>  54
<211>  23
<212>  DNA
<213>  EBV

<400>  54
tctggaccag aaggctccgg cgg                                               23


<210>  55
<211>  23
<212>  DNA
<213>  EBV

<400>  55
gctgccgcgg agggtgatga cgg                                               23


<210>  56
<211>  23
<212>  DNA
<213>  EBV

<400>  56
ggtggcccac cgggtccgct ggg                                               23


<210>  57
<211>  23
<212>  DNA
<213>  EBV

<400>  57
gtcctcgagg gggccgtcgc ggg                                               23


<210>  58
<211>  21
<212>  DNA
<213>  human

<400>  58
aaagaaaata tmmmtggtgt t                                                 21


<210>  59
<211>  21
<212>  DNA
<213>  human

<400>  59
aaagaaaata tctttggtgt t                                                 21


<210>  60
<211>  50
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  GalAcs are conjugated at 3'-end of crRNA via an triazole 
       nNt-Linker.

<400>  60
gcguggccug ggcgggacug guuuuaguac ucuguaauuu uagguaugag                  50


<210>  61
<211>  31
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  crRNA 3'-azide

<400>  61
cuauauggau gaugugguac guuuuagagc u                                      31


<210>  62
<211>  61
<212>  RNA
<213>  Artificial Sequence

<220>
<223>   5'-alkynyl tracrRNA 3'-maleimide.

<400>  62
agcaaguuaa aauaaggcua guccguuauc aacuugaaaa aguggcaccg agucggugcu       60

u                                                                       61


<210>  63
<211>  92
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  lgRNA 3'-maleimide. lgRNA contains a triazole nNt-Linker between 
       u31 and a32.

<400>  63
cuauauggau gaugugguac guuuuagagc uagcaaguua aaauaaggcu aguccguuau       60

caacuugaaa aaguggcacc gagucggugc uu                                     92


<210>  64
<211>  37
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ssDNA, 5'-thiol


<220>
<221>  misc_feature
<222>  (20)..(20)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (24)..(24)
<223>  n is a, c, g, t or u

<400>  64
ucagacttgg cccccatgan tgantgaatc catatag                                37


<210>  65
<211>  129
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA-ssDNA conjugate with a thio-ether nNt-Linker (u93 and u94).


<220>
<221>  misc_feature
<222>  (112)..(112)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (116)..(116)
<223>  n is a, c, g, t or u

<400>  65
cuauauggau gaugugguac guuuuagagc uagcaaguua aaauaaggcu aguccguuau       60

caacuugaaa aaguggcacc gagucggugc uuucagactt ggcccccatg antgantgaa      120

tccatatag                                                              129


<210>  66
<211>  101
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  sgRNA 3'-maleimide.

<400>  66
gcuauaugga ugauguggua cguuuuagag cuagaaauag caaguuaaaa uaaggcuagu       60

ccguuaucaa cuugaaaaag uggcaccgag ucggugcuuu u                          101


<210>  67
<211>  138
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  lgRNA-ssDNA conjugate with a thio-ether nNt-Linker (u101 and 
       u102).


<220>
<221>  misc_feature
<222>  (121)..(121)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (125)..(125)
<223>  n is a, c, g, t or u

<400>  67
gcuauaugga ugauguggua cguuuuagag cuagaaauag caaguuaaaa uaaggcuagu       60

ccguuaucaa cuugaaaaag uggcaccgag ucggugcuuu uucagacttg gcccccatga      120

ntgantgaat ccatatag                                                    138


<210>  68
<211>  129
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ssDNA-lgRNA conjugate with a thio-ether nNt-Linker (u37 and c38).


<220>
<221>  misc_feature
<222>  (19)..(19)
<223>  n is a, c, g, t or u

<220>
<221>  misc_feature
<222>  (23)..(23)
<223>  n is a, c, g, t or u

<400>  68
cagacttggc ccccatgant gantgaatcc atatagucua uauggaugau gugguacguu       60

uuagagcuag caaguuaaaa uaaggcuagu ccguuaucaa cuugaaaaag uggcaccgag      120

ucggugcuu                                                              129


<210>  69
<211>  84
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  cDNA encoding SaCas9 tracrRNA.

<400>  69
gtttacttat acctaaaatt acagaatcta ctaaaacaag gcaaaatgcc gtgtttatct       60

cgtcaacttg ttggcgagat tttt                                              84


<210>  70
<211>  84
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  cDNA encoding SaCas9 tracrRNA.

<400>  70
ggccaaaaat ctcgccaaca agttgacgag ataaacacgg cattttgcct tgttttagta       60

gattctgtaa ttttaggtat aagt                                              84


<210>  71
<211>  4104
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA sequence of engineered SpCas9 protein by site-directed 
       mutagenesis.

<400>  71
tgcgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag      180

gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatcgcc      240

tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aaactggtgg actgcaccga caaggccgac ctgcggctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc      600

atcaacgcct gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga      660

cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac      720

ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg     1020

cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg     1140

gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac     1260

gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc     1320

gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc     1380

agattcgcct ggatgaccag aaagtgcgag gaaaccatca ccccctggaa cttcgaggaa     1440

gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg     1620

agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc     1680

gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc     1740

tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt     1800

atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg     1860

ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc     1920

cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc     1980

aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg     2040

gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg     2160

cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca     2220

gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg     2280

atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga     2340

atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc     2400

gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg     2460

gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat     2520

atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc     2580

gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg     2700

accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag     2760

ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac     2820

actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc     2880

aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag     3000

taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc     3120

aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg     3180

cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt     3240

gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg     3300

cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc     3360

gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc     3420

tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagtgcgtg     3480

aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac     3540

tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag     3600

tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg     3660

cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc     3720

cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag     3900

cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc     3960

cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgac                                            4104


<210>  72
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sequence of an engineered SpCas9-NLS protein.

<400>  72

Cys 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 Ala 
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 Cys 
    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 Cys 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 Cys 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 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Cys Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


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 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Ala Tyr  Pro Tyr Asp Val Pro  Asp Tyr Ala Ser Leu  Gly Ser Gly 
    1370                 1375                 1380             


Ser Pro  Lys Lys Lys Arg Lys  Val Asp 
    1385                 1390         


