                         SEQUENCE LISTING

<110>  INTELLIA THERAPEUTICS, INC.
 
<120>  COMPOSITIONS AND METHODS FOR TTR GENE EDITING AND TREATING ATTR 
       AMYLOIDOSIS

<130>  01155-0013-00PCT

<150>  US 62/566,236
<151>  2017-09-29

<150>  US 62/671,902
<151>  2018-05-15

<160>  278   

<170>  PatentIn version 3.5

<210>  1
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 204, Kozak sequence, and 3 UTR of ALB

<400>  1
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  2
<211>  4403
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript comprising Cas9 ORF corresponding to 
       SEQ ID NO: 205 using codons with generally high expression in 
       humans

<400>  2
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc atgcctaaga aaaagcggaa ggtcgacggg gataagaagt actcaatcgg      120

gctggatatc ggaactaatt ccgtgggttg ggcagtgatc acggatgaat acaaagtgcc      180

gtccaagaag ttcaaggtcc tggggaacac cgatagacac agcatcaaga aaaatctcat      240

cggagccctg ctgtttgact ccggcgaaac cgcagaagcg acccggctca aacgtaccgc      300

gaggcgacgc tacacccggc ggaagaatcg catctgctat ctgcaagaga tcttttcgaa      360

cgaaatggca aaggtcgacg acagcttctt ccaccgcctg gaagaatctt tcctggtgga      420

ggaggacaag aagcatgaac ggcatcctat ctttggaaac atcgtcgacg aagtggcgta      480

ccacgaaaag tacccgacca tctaccatct gcggaagaag ttggttgact caactgacaa      540

ggccgacctc agattgatct acttggccct cgcccatatg atcaaattcc gcggacactt      600

cctgatcgaa ggcgatctga accctgataa ctccgacgtg gataagcttt tcattcaact      660

ggtgcagacc tacaaccaac tgttcgaaga aaacccaatc aatgctagcg gcgtcgatgc      720

caaggccatc ctgtccgccc ggctgtcgaa gtcgcggcgc ctcgaaaacc tgatcgcaca      780

gctgccggga gagaaaaaga acggactttt cggcaacttg atcgctctct cactgggact      840

cactcccaat ttcaagtcca attttgacct ggccgaggac gcgaagctgc aactctcaaa      900

ggacacctac gacgacgact tggacaattt gctggcacaa attggcgatc agtacgcgga      960

tctgttcctt gccgctaaga acctttcgga cgcaatcttg ctgtccgata tcctgcgcgt     1020

gaacaccgaa ataaccaaag cgccgcttag cgcctcgatg attaagcggt acgacgagca     1080

tcaccaggat ctcacgctgc tcaaagcgct cgtgagacag caactgcctg aaaagtacaa     1140

ggagatcttc ttcgaccagt ccaagaatgg gtacgcaggg tacatcgatg gaggcgctag     1200

ccaggaagag ttctataagt tcatcaagcc aatcctggaa aagatggacg gaaccgaaga     1260

actgctggtc aagctgaaca gggaggatct gctccggaaa cagagaacct ttgacaacgg     1320

atccattccc caccagatcc atctgggtga gctgcacgcc atcttgcggc gccaggagga     1380

cttttaccca ttcctcaagg acaaccggga aaagatcgag aaaattctga cgttccgcat     1440

cccgtattac gtgggcccac tggcgcgcgg caattcgcgc ttcgcgtgga tgactagaaa     1500

atcagaggaa accatcactc cttggaattt cgaggaagtt gtggataagg gagcttcggc     1560

acaaagcttc atcgaacgaa tgaccaactt cgacaagaat ctcccaaacg agaaggtgct     1620

tcctaagcac agcctccttt acgaatactt cactgtctac aacgaactga ctaaagtgaa     1680

atacgttact gaaggaatga ggaagccggc ctttctgtcc ggagaacaga agaaagcaat     1740

tgtcgatctg ctgttcaaga ccaaccgcaa ggtgaccgtc aagcagctta aagaggacta     1800

cttcaagaag atcgagtgtt tcgactcagt ggaaatcagc ggggtggagg acagattcaa     1860

cgcttcgctg ggaacctatc atgatctcct gaagatcatc aaggacaagg acttccttga     1920

caacgaggag aacgaggaca tcctggaaga tatcgtcctg accttgaccc ttttcgagga     1980

tcgcgagatg atcgaggaga ggcttaagac ctacgctcat ctcttcgacg ataaggtcat     2040

gaaacaactc aagcgccgcc ggtacactgg ttggggccgc ctctcccgca agctgatcaa     2100

cggtattcgc gataaacaga gcggtaaaac tatcctggat ttcctcaaat cggatggctt     2160

cgctaatcgt aacttcatgc aattgatcca cgacgacagc ctgaccttta aggaggacat     2220

ccaaaaagca caagtgtccg gacagggaga ctcactccat gaacacatcg cgaatctggc     2280

cggttcgccg gcgattaaga agggaattct gcaaactgtg aaggtggtcg acgagctggt     2340

gaaggtcatg ggacggcaca aaccggagaa tatcgtgatt gaaatggccc gagaaaacca     2400

gactacccag aagggccaga aaaactcccg cgaaaggatg aagcggatcg aagaaggaat     2460

caaggagctg ggcagccaga tcctgaaaga gcacccggtg gaaaacacgc agctgcagaa     2520

cgagaagctc tacctgtact atttgcaaaa tggacgggac atgtacgtgg accaagagct     2580

ggacatcaat cggttgtctg attacgacgt ggaccacatc gttccacagt cctttctgaa     2640

ggatgactcg atcgataaca aggtgttgac tcgcagcgac aagaacagag ggaagtcaga     2700

taatgtgcca tcggaggagg tcgtgaagaa gatgaagaat tactggcggc agctcctgaa     2760

tgcgaagctg attacccaga gaaagtttga caatctcact aaagccgagc gcggcggact     2820

ctcagagctg gataaggctg gattcatcaa acggcagctg gtcgagactc ggcagattac     2880

caagcacgtg gcgcagatct tggactcccg catgaacact aaatacgacg agaacgataa     2940

gctcatccgg gaagtgaagg tgattaccct gaaaagcaaa cttgtgtcgg actttcggaa     3000

ggactttcag ttttacaaag tgagagaaat caacaactac catcacgcgc atgacgcata     3060

cctcaacgct gtggtcggta ccgccctgat caaaaagtac cctaaacttg aatcggagtt     3120

tgtgtacgga gactacaagg tctacgacgt gaggaagatg atagccaagt ccgaacagga     3180

aatcgggaaa gcaactgcga aatacttctt ttactcaaac atcatgaact ttttcaagac     3240

tgaaattacg ctggccaatg gagaaatcag gaagaggcca ctgatcgaaa ctaacggaga     3300

aacgggcgaa atcgtgtggg acaagggcag ggacttcgca actgttcgca aagtgctctc     3360

tatgccgcaa gtcaatattg tgaagaaaac cgaagtgcaa accggcggat tttcaaagga     3420

atcgatcctc ccaaagagaa atagcgacaa gctcattgca cgcaagaaag actgggaccc     3480

gaagaagtac ggaggattcg attcgccgac tgtcgcatac tccgtcctcg tggtggccaa     3540

ggtggagaag ggaaagagca aaaagctcaa atccgtcaaa gagctgctgg ggattaccat     3600

catggaacga tcctcgttcg agaagaaccc gattgatttc ctcgaggcga agggttacaa     3660

ggaggtgaag aaggatctga tcatcaaact ccccaagtac tcactgttcg aactggaaaa     3720

tggtcggaag cgcatgctgg cttcggccgg agaactccaa aaaggaaatg agctggcctt     3780

gcctagcaag tacgtcaact tcctctatct tgcttcgcac tacgaaaaac tcaaagggtc     3840

accggaagat aacgaacaga agcagctttt cgtggagcag cacaagcatt atctggatga     3900

aatcatcgaa caaatctccg agttttcaaa gcgcgtgatc ctcgccgacg ccaacctcga     3960

caaagtcctg tcggcctaca ataagcatag agataagccg atcagagaac aggccgagaa     4020

cattatccac ttgttcaccc tgactaacct gggagcccca gccgccttca agtacttcga     4080

tactactatc gatcgcaaaa gatacacgtc caccaaggaa gttctggacg cgaccctgat     4140

ccaccaaagc atcactggac tctacgaaac taggatcgat ctgtcgcagc tgggtggcga     4200

ttgatagtct agccatcaca tttaaaagca tctcagccta ccatgagaat aagagaaaga     4260

aaatgaagat caatagctta ttcatctctt tttctttttc gttggtgtaa agccaacacc     4320

ctgtctaaaa aacataaatt tctttaatca ttttgcctct tttctctgtg cttcaattaa     4380

taaaaaatgg aaagaacctc gag                                             4403


<210>  3
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: modified sgRNA sequence


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

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

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  3
nnnnnnnnnn nnnnnnnnnn guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  4
<211>  105
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: 30/30/39 poly-A sequence

<400>  4
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa gcgaaaaaaa aaaaaaaaaa aaaaaaaaaa       60

aaaccgaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa                      105


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

<220>
<223>  Synthetic: CR003335 gRNA targeting Human TTR (Exon 1)

<400>  5
cugcuccucc ucugccuugc                                                   20


<210>  6
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003336 gRNA targeting Human TTR (Exon 1)

<400>  6
ccuccucugc cuugcuggac                                                   20


<210>  7
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003337 gRNA targeting Human TTR (Exon 1)

<400>  7
ccaguccagc aaggcagagg                                                   20


<210>  8
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003338 gRNA targeting Human TTR (Exon 1)

<400>  8
auaccagucc agcaaggcag                                                   20


<210>  9
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003339 gRNA targeting Human TTR (Exon 1)

<400>  9
acacaaauac caguccagca                                                   20


<210>  10
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003340 gRNA targeting Human TTR (Exon 1)

<400>  10
uggacuggua uuugugucug                                                   20


<210>  11
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003341 gRNA targeting Human TTR (Exon 1)

<400>  11
cugguauuug ugucugaggc                                                   20


<210>  12
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003342 gRNA targeting Human TTR (Exon 2)

<400>  12
cuucucuaca cccagggcac                                                   20


<210>  13
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003343 gRNA targeting Human TTR (Exon 2)

<400>  13
cagaggacac uuggauucac                                                   20


<210>  14
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003344 gRNA targeting Human TTR (Exon 2)

<400>  14
uuugaccauc agaggacacu                                                   20


<210>  15
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003345 gRNA targeting Human TTR (Exon 2)

<400>  15
ucuagaacuu ugaccaucag                                                   20


<210>  16
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003346 gRNA targeting Human TTR (Exon 2)

<400>  16
aaaguucuag augcuguccg                                                   20


<210>  17
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003347 gRNA targeting Human TTR (Exon 2)

<400>  17
cauugauggc aggacugccu                                                   20


<210>  18
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003348 gRNA targeting Human TTR (Exon 2)

<400>  18
aggcaguccu gccaucaaug                                                   20


<210>  19
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003349 gRNA targeting Human TTR (Exon 2)

<400>  19
ugcacggcca cauugauggc                                                   20


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

<220>
<223>  Synthetic: CR003350 gRNA targeting Human TTR (Exon 2)

<400>  20
cacaugcacg gccacauuga                                                   20


<210>  21
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003351 gRNA targeting Human TTR (Exon 2)

<400>  21
agccuuucug aacacaugca                                                   20


<210>  22
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003352 gRNA targeting Human TTR (Exon 2)

<400>  22
gaaaggcugc ugaugacacc                                                   20


<210>  23
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003353 gRNA targeting Human TTR (Exon 2)

<400>  23
aaaggcugcu gaugacaccu                                                   20


<210>  24
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003354 gRNA targeting Human TTR (Exon 2)

<400>  24
accugggagc cauuugccuc                                                   20


<210>  25
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003355 gRNA targeting Human TTR (Exon 2)

<400>  25
cccagaggca aauggcuccc                                                   20


<210>  26
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003356 gRNA targeting Human TTR (Exon 2)

<400>  26
gcaacuuacc cagaggcaaa                                                   20


<210>  27
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003357 gRNA targeting Human TTR (Exon 2)

<400>  27
uucuuuggca acuuacccag                                                   20


<210>  28
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003358 gRNA targeting Human TTR (Exon 3)

<400>  28
augcagcucu ccagacucac                                                   20


<210>  29
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003359 gRNA targeting Human TTR (Exon 3)

<400>  29
agugagucug gagagcugca                                                   20


<210>  30
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003360 gRNA targeting Human TTR (Exon 3)

<400>  30
gugagucugg agagcugcau                                                   20


<210>  31
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003361 gRNA targeting Human TTR (Exon 3)

<400>  31
gcugcauggg cucacaacug                                                   20


<210>  32
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003362 gRNA targeting Human TTR (Exon 3)

<400>  32
gcaugggcuc acaacugagg                                                   20


<210>  33
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003363 gRNA targeting Human TTR (Exon 3)

<400>  33
acugaggagg aauuuguaga                                                   20


<210>  34
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003364 gRNA targeting Human TTR (Exon 3)

<400>  34
cugaggagga auuuguagaa                                                   20


<210>  35
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003365 gRNA targeting Human TTR (Exon 3)

<400>  35
uguagaaggg auauacaaag                                                   20


<210>  36
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003366 gRNA targeting Human TTR (Exon 3)

<400>  36
aaauagacac caaaucuuac                                                   20


<210>  37
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003367 gRNA targeting Human TTR (Exon 3)

<400>  37
agacaccaaa ucuuacugga                                                   20


<210>  38
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003368 gRNA targeting Human TTR (Exon 3)

<400>  38
aagugccuuc caguaagauu                                                   20


<210>  39
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003369 gRNA targeting Human TTR (Exon 3)

<400>  39
cucugcaugc ucauggaaug                                                   20


<210>  40
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003370 gRNA targeting Human TTR (Exon 3)

<400>  40
ccucugcaug cucauggaau                                                   20


<210>  41
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003371 gRNA targeting Human TTR (Exon 3)

<400>  41
accucugcau gcucauggaa                                                   20


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

<220>
<223>  Synthetic: CR003372 gRNA targeting Human TTR (Exon 3)

<400>  42
uacucaccuc ugcaugcuca                                                   20


<210>  43
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003373 gRNA targeting Human TTR (Exon 4)

<400>  43
guauucacag ccaacgacuc                                                   20


<210>  44
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003374 gRNA targeting Human TTR (Exon 4)

<400>  44
gcggcggggg ccggagucgu                                                   20


<210>  45
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003375 gRNA targeting Human TTR (Exon 4)

<400>  45
aaugguguag cggcgggggc                                                   20


<210>  46
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003376 gRNA targeting Human TTR (Exon 4)

<400>  46
cggcaauggu guagcggcgg                                                   20


<210>  47
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003377 gRNA targeting Human TTR (Exon 4)

<400>  47
gcggcaaugg uguagcggcg                                                   20


<210>  48
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003378 gRNA targeting Human TTR (Exon 4)

<400>  48
ggcggcaaug guguagcggc                                                   20


<210>  49
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003379 gRNA targeting Human TTR (Exon 4)

<400>  49
gggcggcaau gguguagcgg                                                   20


<210>  50
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003380 gRNA targeting Human TTR (Exon 4)

<400>  50
gcagggcggc aaugguguag                                                   20


<210>  51
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003381 gRNA targeting Human TTR (Exon 4)

<400>  51
ggggcucagc agggcggcaa                                                   20


<210>  52
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003382 gRNA targeting Human TTR (Exon 4)

<400>  52
ggaguagggg cucagcaggg                                                   20


<210>  53
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003383 gRNA targeting Human TTR (Exon 4)

<400>  53
auaggaguag gggcucagca                                                   20


<210>  54
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003384 gRNA targeting Human TTR (Exon 4)

<400>  54
aauaggagua ggggcucagc                                                   20


<210>  55
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003385 gRNA targeting Human TTR (Exon 4)

<400>  55
ccccuacucc uauuccacca                                                   20


<210>  56
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003386 gRNA targeting Human TTR (Exon 4)

<400>  56
ccguggugga auaggaguag                                                   20


<210>  57
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003387 gRNA targeting Human TTR (Exon 4)

<400>  57
gccguggugg aauaggagua                                                   20


<210>  58
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003388 gRNA targeting Human TTR (Exon 4)

<400>  58
gacgacagcc gugguggaau                                                   20


<210>  59
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003389 gRNA targeting Human TTR (Exon 4)

<400>  59
auuggugacg acagccgugg                                                   20


<210>  60
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003390 gRNA targeting Human TTR (Exon 4)

<400>  60
gggauuggug acgacagccg                                                   20


<210>  61
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003391 gRNA targeting Human TTR (Exon 4)

<400>  61
ggcugucguc accaauccca                                                   20


<210>  62
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR003392 gRNA targeting Human TTR (Exon 4)

<400>  62
agucccucau uccuugggau                                                   20


<210>  63
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005298 gRNA targeting Human TTR (Exon 1)

<400>  63
uccacucauu cuuggcagga                                                   20


<210>  64
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005299 gRNA targeting Human TTR (Exon 4)

<400>  64
agccguggug gaauaggagu                                                   20


<210>  65
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005300 gRNA targeting Human TTR (Exon 1)

<400>  65
ucacagaaac acucaccgua                                                   20


<210>  66
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005301 gRNA targeting Human TTR (Exon 1)

<400>  66
gucacagaaa cacucaccgu                                                   20


<210>  67
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005302 gRNA targeting Human TTR (Exon 2)

<400>  67
acgugucuuc ucuacaccca                                                   20


<210>  68
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005303 gRNA targeting Human TTR (Exon 2)

<400>  68
ugaauccaag uguccucuga                                                   20


<210>  69
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005304 gRNA targeting Human TTR (Exon 2)

<400>  69
ggccgugcau guguucagaa                                                   20


<210>  70
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005305 gRNA targeting Human TTR (Exon 3)

<400>  70
uauaggaaaa ccagugaguc                                                   20


<210>  71
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005306 gRNA targeting Human TTR (Exon 3)

<400>  71
aaaucuuacu ggaaggcacu                                                   20


<210>  72
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005307 gRNA targeting Human TTR (Exon 4)

<400>  72
ugucugucuu cucucauagg                                                   20


<210>  73
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR000689 gRNA targeting Cyno TTR

<400>  73
acacaaauac caguccagcg                                                   20


<210>  74
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005364 gRNA targeting Cyno TTR

<400>  74
aaaggcugcu gaugagaccu                                                   20


<210>  75
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005365 gRNA targeting Cyno TTR

<400>  75
cauugacagc aggacugccu                                                   20


<210>  76
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005366 gRNA targeting Cyno TTR

<400>  76
auaccagucc agcgaggcag                                                   20


<210>  77
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005367 gRNA targeting Cyno TTR

<400>  77
ccaguccagc gaggcagagg                                                   20


<210>  78
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005368 gRNA targeting Cyno TTR

<400>  78
ccuccucugc cucgcuggac                                                   20


<210>  79
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005369 gRNA targeting Cyno TTR

<400>  79
aaaguucuag augccguccg                                                   20


<210>  80
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005370 gRNA targeting Cyno TTR

<400>  80
acuugucuuc ucuauaccca                                                   20


<210>  81
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005371 gRNA targeting Cyno TTR

<400>  81
aagugacuuc caguaagauu                                                   20


<210>  82
<211>  20
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CR005372 gRNA targeting Cyno TTR

<400>  82
aaaaggcugc ugaugagacc                                                   20


<210>  83

<400>  83
000

<210>  84

<400>  84
000

<210>  85

<400>  85
000

<210>  86

<400>  86
000

<210>  87
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000480 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  87
aaaggcugcu gaugacaccu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  88
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000481 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  88
ucuagaacuu ugaccaucag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  89
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000482 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  89
uguagaaggg auauacaaag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  90
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000483 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  90
uccacucauu cuuggcagga guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  91
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000484 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  91
agacaccaaa ucuuacugga guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  92
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000485 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  92
ccuccucugc cuugcuggac guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  93
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000486 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  93
acacaaauac caguccagca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  94
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000487 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  94
uucuuuggca acuuacccag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  95
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000488 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  95
aaaguucuag augcuguccg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  96
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000489 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  96
uuugaccauc agaggacacu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  97
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000490 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  97
aaauagacac caaaucuuac guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  98
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000491 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  98
auaccagucc agcaaggcag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  99
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000492 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  99
cuucucuaca cccagggcac guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


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

<220>
<223>  Synthetic: G000493 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  100
aagugccuuc caguaagauu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  101
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000494 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  101
gugagucugg agagcugcau guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  102
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000495 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  102
cagaggacac uuggauucac guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  103
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000496 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  103
ggccgugcau guguucagaa guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  104
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000497 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  104
cugcuccucc ucugccuugc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  105
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000498 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  105
agugagucug gagagcugca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  106
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000499 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  106
ugaauccaag uguccucuga guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  107
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000500 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  107
ccaguccagc aaggcagagg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  108
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000501 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  108
ucacagaaac acucaccgua guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  109
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000567 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  109
gaaaggcugc ugaugacacc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  110
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000568 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  110
ggcugucguc accaauccca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  111
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000570 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  111
cauugauggc aggacugccu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  112
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000571 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  112
gucacagaaa cacucaccgu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  113
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000572 sgRNA modified sequence targeting Human TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  113
ccccuacucc uauuccacca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  114
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000502 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  114
acacaaauac caguccagcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  115
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000503 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  115
aaaaggcugc ugaugagacc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  116
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000504 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  116
aaaggcugcu gaugagaccu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  117
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000505 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  117
cauugacagc aggacugccu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  118
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000506 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  118
auaccagucc agcgaggcag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  119
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000507 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  119
ccaguccagc gaggcagagg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  120
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000508 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  120
ccuccucugc cucgcuggac guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  121
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000509 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  121
aaaguucuag augccguccg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  122
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000510 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  122
acuugucuuc ucuauaccca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  123
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000511 sgRNA modified sequence targeting Cyno TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  123
aagugacuuc caguaagauu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  124
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000282 sgRNA modified sequence targeting Mouse TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  124
uuacagccac gucuacagca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  125

<400>  125
000

<210>  126

<400>  126
000

<210>  127

<400>  127
000

<210>  128

<400>  128
000

<210>  129

<400>  129
000

<210>  130

<400>  130
000

<210>  131

<400>  131
000

<210>  132

<400>  132
000

<210>  133

<400>  133
000

<210>  134

<400>  134
000

<210>  135

<400>  135
000

<210>  136

<400>  136
000

<210>  137

<400>  137
000

<210>  138

<400>  138
000

<210>  139

<400>  139
000

<210>  140

<400>  140
000

<210>  141

<400>  141
000

<210>  142

<400>  142
000

<210>  143

<400>  143
000

<210>  144

<400>  144
000

<210>  145

<400>  145
000

<210>  146

<400>  146
000

<210>  147

<400>  147
000

<210>  148

<400>  148
000

<210>  149

<400>  149
000

<210>  150

<400>  150
000

<210>  151

<400>  151
000

<210>  152

<400>  152
000

<210>  153

<400>  153
000

<210>  154

<400>  154
000

<210>  155

<400>  155
000

<210>  156

<400>  156
000

<210>  157

<400>  157
000

<210>  158

<400>  158
000

<210>  159

<400>  159
000

<210>  160

<400>  160
000

<210>  161

<400>  161
000

<210>  162

<400>  162
000

<210>  163

<400>  163
000

<210>  164

<400>  164
000

<210>  165

<400>  165
000

<210>  166

<400>  166
000

<210>  167

<400>  167
000

<210>  168

<400>  168
000

<210>  169

<400>  169
000

<210>  170

<400>  170
000

<210>  171

<400>  171
000

<210>  172

<400>  172
000

<210>  173

<400>  173
000

<210>  174

<400>  174
000

<210>  175

<400>  175
000

<210>  176

<400>  176
000

<210>  177

<400>  177
000

<210>  178

<400>  178
000

<210>  179

<400>  179
000

<210>  180

<400>  180
000

<210>  181

<400>  181
000

<210>  182

<400>  182
000

<210>  183

<400>  183
000

<210>  184

<400>  184
000

<210>  185

<400>  185
000

<210>  186

<400>  186
000

<210>  187

<400>  187
000

<210>  188

<400>  188
000

<210>  189

<400>  189
000

<210>  190

<400>  190
000

<210>  191

<400>  191
000

<210>  192

<400>  192
000

<210>  193

<400>  193
000

<210>  194

<400>  194
000

<210>  195

<400>  195
000

<210>  196

<400>  196
000

<210>  197

<400>  197
000

<210>  198

<400>  198
000

<210>  199

<400>  199
000

<210>  200

<400>  200
000

<210>  201
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: DNA coding sequence of Cas9 using the thymidine analog
       of the minimal uridine codons listed in Table 3, with start and 
       stop codons

<400>  201
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcccga agaagaagag aaaggtctag     4140


<210>  202
<211>  4143
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: DNA coding sequence of Cas9 using codons with 
       generally high expression in humans

<400>  202
atggataaga agtactcaat cgggctggat atcggaacta attccgtggg ttgggcagtg       60

atcacggatg aatacaaagt gccgtccaag aagttcaagg tcctggggaa caccgataga      120

cacagcatca agaaaaatct catcggagcc ctgctgtttg actccggcga aaccgcagaa      180

gcgacccggc tcaaacgtac cgcgaggcga cgctacaccc ggcggaagaa tcgcatctgc      240

tatctgcaag agatcttttc gaacgaaatg gcaaaggtcg acgacagctt cttccaccgc      300

ctggaagaat ctttcctggt ggaggaggac aagaagcatg aacggcatcc tatctttgga      360

aacatcgtcg acgaagtggc gtaccacgaa aagtacccga ccatctacca tctgcggaag      420

aagttggttg actcaactga caaggccgac ctcagattga tctacttggc cctcgcccat      480

atgatcaaat tccgcggaca cttcctgatc gaaggcgatc tgaaccctga taactccgac      540

gtggataagc ttttcattca actggtgcag acctacaacc aactgttcga agaaaaccca      600

atcaatgcta gcggcgtcga tgccaaggcc atcctgtccg cccggctgtc gaagtcgcgg      660

cgcctcgaaa acctgatcgc acagctgccg ggagagaaaa agaacggact tttcggcaac      720

ttgatcgctc tctcactggg actcactccc aatttcaagt ccaattttga cctggccgag      780

gacgcgaagc tgcaactctc aaaggacacc tacgacgacg acttggacaa tttgctggca      840

caaattggcg atcagtacgc ggatctgttc cttgccgcta agaacctttc ggacgcaatc      900

ttgctgtccg atatcctgcg cgtgaacacc gaaataacca aagcgccgct tagcgcctcg      960

atgattaagc ggtacgacga gcatcaccag gatctcacgc tgctcaaagc gctcgtgaga     1020

cagcaactgc ctgaaaagta caaggagatc ttcttcgacc agtccaagaa tgggtacgca     1080

gggtacatcg atggaggcgc tagccaggaa gagttctata agttcatcaa gccaatcctg     1140

gaaaagatgg acggaaccga agaactgctg gtcaagctga acagggagga tctgctccgg     1200

aaacagagaa cctttgacaa cggatccatt ccccaccaga tccatctggg tgagctgcac     1260

gccatcttgc ggcgccagga ggacttttac ccattcctca aggacaaccg ggaaaagatc     1320

gagaaaattc tgacgttccg catcccgtat tacgtgggcc cactggcgcg cggcaattcg     1380

cgcttcgcgt ggatgactag aaaatcagag gaaaccatca ctccttggaa tttcgaggaa     1440

gttgtggata agggagcttc ggcacaaagc ttcatcgaac gaatgaccaa cttcgacaag     1500

aatctcccaa acgagaaggt gcttcctaag cacagcctcc tttacgaata cttcactgtc     1560

tacaacgaac tgactaaagt gaaatacgtt actgaaggaa tgaggaagcc ggcctttctg     1620

tccggagaac agaagaaagc aattgtcgat ctgctgttca agaccaaccg caaggtgacc     1680

gtcaagcagc ttaaagagga ctacttcaag aagatcgagt gtttcgactc agtggaaatc     1740

agcggggtgg aggacagatt caacgcttcg ctgggaacct atcatgatct cctgaagatc     1800

atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctgga agatatcgtc     1860

ctgaccttga cccttttcga ggatcgcgag atgatcgagg agaggcttaa gacctacgct     1920

catctcttcg acgataaggt catgaaacaa ctcaagcgcc gccggtacac tggttggggc     1980

cgcctctccc gcaagctgat caacggtatt cgcgataaac agagcggtaa aactatcctg     2040

gatttcctca aatcggatgg cttcgctaat cgtaacttca tgcaattgat ccacgacgac     2100

agcctgacct ttaaggagga catccaaaaa gcacaagtgt ccggacaggg agactcactc     2160

catgaacaca tcgcgaatct ggccggttcg ccggcgatta agaagggaat tctgcaaact     2220

gtgaaggtgg tcgacgagct ggtgaaggtc atgggacggc acaaaccgga gaatatcgtg     2280

attgaaatgg cccgagaaaa ccagactacc cagaagggcc agaaaaactc ccgcgaaagg     2340

atgaagcgga tcgaagaagg aatcaaggag ctgggcagcc agatcctgaa agagcacccg     2400

gtggaaaaca cgcagctgca gaacgagaag ctctacctgt actatttgca aaatggacgg     2460

gacatgtacg tggaccaaga gctggacatc aatcggttgt ctgattacga cgtggaccac     2520

atcgttccac agtcctttct gaaggatgac tcgatcgata acaaggtgtt gactcgcagc     2580

gacaagaaca gagggaagtc agataatgtg ccatcggagg aggtcgtgaa gaagatgaag     2640

aattactggc ggcagctcct gaatgcgaag ctgattaccc agagaaagtt tgacaatctc     2700

actaaagccg agcgcggcgg actctcagag ctggataagg ctggattcat caaacggcag     2760

ctggtcgaga ctcggcagat taccaagcac gtggcgcaga tcttggactc ccgcatgaac     2820

actaaatacg acgagaacga taagctcatc cgggaagtga aggtgattac cctgaaaagc     2880

aaacttgtgt cggactttcg gaaggacttt cagttttaca aagtgagaga aatcaacaac     2940

taccatcacg cgcatgacgc atacctcaac gctgtggtcg gtaccgccct gatcaaaaag     3000

taccctaaac ttgaatcgga gtttgtgtac ggagactaca aggtctacga cgtgaggaag     3060

atgatagcca agtccgaaca ggaaatcggg aaagcaactg cgaaatactt cttttactca     3120

aacatcatga actttttcaa gactgaaatt acgctggcca atggagaaat caggaagagg     3180

ccactgatcg aaactaacgg agaaacgggc gaaatcgtgt gggacaaggg cagggacttc     3240

gcaactgttc gcaaagtgct ctctatgccg caagtcaata ttgtgaagaa aaccgaagtg     3300

caaaccggcg gattttcaaa ggaatcgatc ctcccaaaga gaaatagcga caagctcatt     3360

gcacgcaaga aagactggga cccgaagaag tacggaggat tcgattcgcc gactgtcgca     3420

tactccgtcc tcgtggtggc caaggtggag aagggaaaga gcaaaaagct caaatccgtc     3480

aaagagctgc tggggattac catcatggaa cgatcctcgt tcgagaagaa cccgattgat     3540

ttcctcgagg cgaagggtta caaggaggtg aagaaggatc tgatcatcaa actccccaag     3600

tactcactgt tcgaactgga aaatggtcgg aagcgcatgc tggcttcggc cggagaactc     3660

caaaaaggaa atgagctggc cttgcctagc aagtacgtca acttcctcta tcttgcttcg     3720

cactacgaaa aactcaaagg gtcaccggaa gataacgaac agaagcagct tttcgtggag     3780

cagcacaagc attatctgga tgaaatcatc gaacaaatct ccgagttttc aaagcgcgtg     3840

atcctcgccg acgccaacct cgacaaagtc ctgtcggcct acaataagca tagagataag     3900

ccgatcagag aacaggccga gaacattatc cacttgttca ccctgactaa cctgggagcc     3960

ccagccgcct tcaagtactt cgatactact atcgatcgca aaagatacac gtccaccaag     4020

gaagttctgg acgcgaccct gatccaccaa agcatcactg gactctacga aactaggatc     4080

gatctgtcgc agctgggtgg cgatggcggt ggatctccga aaaagaagag aaaggtgtaa     4140

tga                                                                   4143


<210>  203
<211>  1379
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 with one nuclear 
       localization signal (1xNLS) as the C-terminal 7 amino acids

<400>  203

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    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             


Ser 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             


Gly Gly  Gly Ser Pro Lys Lys  Lys Arg Lys Val 
    1370                 1375                 


<210>  204
<211>  4140
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF using minimal uridine codons, with start
       and stop codons

<400>  204
auggacaaga aguacagcau cggacuggac aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucuag     4140


<210>  205
<211>  4143
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF using codons with generally high 
       expression in humans, with start and stop codons

<400>  205
auggauaaga aguacucaau cgggcuggau aucggaacua auuccguggg uugggcagug       60

aucacggaug aauacaaagu gccguccaag aaguucaagg uccuggggaa caccgauaga      120

cacagcauca agaaaaaucu caucggagcc cugcuguuug acuccggcga aaccgcagaa      180

gcgacccggc ucaaacguac cgcgaggcga cgcuacaccc ggcggaagaa ucgcaucugc      240

uaucugcaag agaucuuuuc gaacgaaaug gcaaaggucg acgacagcuu cuuccaccgc      300

cuggaagaau cuuuccuggu ggaggaggac aagaagcaug aacggcaucc uaucuuugga      360

aacaucgucg acgaaguggc guaccacgaa aaguacccga ccaucuacca ucugcggaag      420

aaguugguug acucaacuga caaggccgac cucagauuga ucuacuuggc ccucgcccau      480

augaucaaau uccgcggaca cuuccugauc gaaggcgauc ugaacccuga uaacuccgac      540

guggauaagc uuuucauuca acuggugcag accuacaacc aacuguucga agaaaaccca      600

aucaaugcua gcggcgucga ugccaaggcc auccuguccg cccggcuguc gaagucgcgg      660

cgccucgaaa accugaucgc acagcugccg ggagagaaaa agaacggacu uuucggcaac      720

uugaucgcuc ucucacuggg acucacuccc aauuucaagu ccaauuuuga ccuggccgag      780

gacgcgaagc ugcaacucuc aaaggacacc uacgacgacg acuuggacaa uuugcuggca      840

caaauuggcg aucaguacgc ggaucuguuc cuugccgcua agaaccuuuc ggacgcaauc      900

uugcuguccg auauccugcg cgugaacacc gaaauaacca aagcgccgcu uagcgccucg      960

augauuaagc gguacgacga gcaucaccag gaucucacgc ugcucaaagc gcucgugaga     1020

cagcaacugc cugaaaagua caaggagauc uucuucgacc aguccaagaa uggguacgca     1080

ggguacaucg auggaggcgc uagccaggaa gaguucuaua aguucaucaa gccaauccug     1140

gaaaagaugg acggaaccga agaacugcug gucaagcuga acagggagga ucugcuccgg     1200

aaacagagaa ccuuugacaa cggauccauu ccccaccaga uccaucuggg ugagcugcac     1260

gccaucuugc ggcgccagga ggacuuuuac ccauuccuca aggacaaccg ggaaaagauc     1320

gagaaaauuc ugacguuccg caucccguau uacgugggcc cacuggcgcg cggcaauucg     1380

cgcuucgcgu ggaugacuag aaaaucagag gaaaccauca cuccuuggaa uuucgaggaa     1440

guuguggaua agggagcuuc ggcacaaagc uucaucgaac gaaugaccaa cuucgacaag     1500

aaucucccaa acgagaaggu gcuuccuaag cacagccucc uuuacgaaua cuucacuguc     1560

uacaacgaac ugacuaaagu gaaauacguu acugaaggaa ugaggaagcc ggccuuucug     1620

uccggagaac agaagaaagc aauugucgau cugcuguuca agaccaaccg caaggugacc     1680

gucaagcagc uuaaagagga cuacuucaag aagaucgagu guuucgacuc aguggaaauc     1740

agcggggugg aggacagauu caacgcuucg cugggaaccu aucaugaucu ccugaagauc     1800

aucaaggaca aggacuuccu ugacaacgag gagaacgagg acauccugga agauaucguc     1860

cugaccuuga cccuuuucga ggaucgcgag augaucgagg agaggcuuaa gaccuacgcu     1920

caucucuucg acgauaaggu caugaaacaa cucaagcgcc gccgguacac ugguuggggc     1980

cgccucuccc gcaagcugau caacgguauu cgcgauaaac agagcgguaa aacuauccug     2040

gauuuccuca aaucggaugg cuucgcuaau cguaacuuca ugcaauugau ccacgacgac     2100

agccugaccu uuaaggagga cauccaaaaa gcacaagugu ccggacaggg agacucacuc     2160

caugaacaca ucgcgaaucu ggccgguucg ccggcgauua agaagggaau ucugcaaacu     2220

gugaaggugg ucgacgagcu ggugaagguc augggacggc acaaaccgga gaauaucgug     2280

auugaaaugg cccgagaaaa ccagacuacc cagaagggcc agaaaaacuc ccgcgaaagg     2340

augaagcgga ucgaagaagg aaucaaggag cugggcagcc agauccugaa agagcacccg     2400

guggaaaaca cgcagcugca gaacgagaag cucuaccugu acuauuugca aaauggacgg     2460

gacauguacg uggaccaaga gcuggacauc aaucgguugu cugauuacga cguggaccac     2520

aucguuccac aguccuuucu gaaggaugac ucgaucgaua acaagguguu gacucgcagc     2580

gacaagaaca gagggaaguc agauaaugug ccaucggagg aggucgugaa gaagaugaag     2640

aauuacuggc ggcagcuccu gaaugcgaag cugauuaccc agagaaaguu ugacaaucuc     2700

acuaaagccg agcgcggcgg acucucagag cuggauaagg cuggauucau caaacggcag     2760

cuggucgaga cucggcagau uaccaagcac guggcgcaga ucuuggacuc ccgcaugaac     2820

acuaaauacg acgagaacga uaagcucauc cgggaaguga aggugauuac ccugaaaagc     2880

aaacuugugu cggacuuucg gaaggacuuu caguuuuaca aagugagaga aaucaacaac     2940

uaccaucacg cgcaugacgc auaccucaac gcuguggucg guaccgcccu gaucaaaaag     3000

uacccuaaac uugaaucgga guuuguguac ggagacuaca aggucuacga cgugaggaag     3060

augauagcca aguccgaaca ggaaaucggg aaagcaacug cgaaauacuu cuuuuacuca     3120

aacaucauga acuuuuucaa gacugaaauu acgcuggcca auggagaaau caggaagagg     3180

ccacugaucg aaacuaacgg agaaacgggc gaaaucgugu gggacaaggg cagggacuuc     3240

gcaacuguuc gcaaagugcu cucuaugccg caagucaaua uugugaagaa aaccgaagug     3300

caaaccggcg gauuuucaaa ggaaucgauc cucccaaaga gaaauagcga caagcucauu     3360

gcacgcaaga aagacuggga cccgaagaag uacggaggau ucgauucgcc gacugucgca     3420

uacuccgucc ucgugguggc caagguggag aagggaaaga gcaaaaagcu caaauccguc     3480

aaagagcugc uggggauuac caucauggaa cgauccucgu ucgagaagaa cccgauugau     3540

uuccucgagg cgaaggguua caaggaggug aagaaggauc ugaucaucaa acuccccaag     3600

uacucacugu ucgaacugga aaauggucgg aagcgcaugc uggcuucggc cggagaacuc     3660

caaaaaggaa augagcuggc cuugccuagc aaguacguca acuuccucua ucuugcuucg     3720

cacuacgaaa aacucaaagg gucaccggaa gauaacgaac agaagcagcu uuucguggag     3780

cagcacaagc auuaucugga ugaaaucauc gaacaaaucu ccgaguuuuc aaagcgcgug     3840

auccucgccg acgccaaccu cgacaaaguc cugucggccu acaauaagca uagagauaag     3900

ccgaucagag aacaggccga gaacauuauc cacuuguuca cccugacuaa ccugggagcc     3960

ccagccgccu ucaaguacuu cgauacuacu aucgaucgca aaagauacac guccaccaag     4020

gaaguucugg acgcgacccu gauccaccaa agcaucacug gacucuacga aacuaggauc     4080

gaucugucgc agcugggugg cgauggcggu ggaucuccga aaaagaagag aaagguguaa     4140

uga                                                                   4143


<210>  206
<211>  1379
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase (D10A) amino acid sequence

<400>  206

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    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             


Ser 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             


Gly Gly  Gly Ser Pro Lys Lys  Lys Arg Lys Val 
    1370                 1375                 


<210>  207
<211>  4140
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase (D10A) mRNA ORF

<400>  207
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucuag     4140


<210>  208
<211>  1379
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 (D10A H840A) amino acid sequence

<400>  208

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp Ala 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             


Ser 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             


Gly Gly  Gly Ser Pro Lys Lys  Lys Arg Lys Val 
    1370                 1375                 


<210>  209
<211>  4140
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 (D10A H840A) mRNA ORF

<400>  209
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgacgca     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucuag     4140


<210>  210
<211>  4134
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA coding sequence using minimal uridine codons
       (no start or stop codons; suitable for inclusion in fusion 
       protein coding sequence)

<400>  210
gacaagaagu acagcaucgg acuggacauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa gguc           4134


<210>  211
<211>  4134
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase bare coding sequence

<400>  211
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa gguc           4134


<210>  212
<211>  4134
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 bare coding sequence

<400>  212
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgacgcaauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa gguc           4134


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

<220>
<223>  Synthetic: Amino acid sequence of Cas9 (without NLS)

<400>  213

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    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             


Ser 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             


<210>  214
<211>  4107
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF encoding SEQ ID NO: 213 using minimal 
       uridine codons, with start and stop codons

<400>  214
auggacaaga aguacagcau cggacuggac aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacuag                                         4107


<210>  215
<211>  4101
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 coding sequence encoding SEQ ID NO: 213 using 
       minimal uridine codons (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  215
gacaagaagu acagcaucgg acuggacauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga c                                               4101


<210>  216
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 nickase (without NLS)

<400>  216

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    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             


Ser 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             


<210>  217
<211>  4107
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase mRNA ORF encoding SEQ ID NO: 216 using 
       minimal uridine codons as listed in Table 3, with start and stop 
       codons

<400>  217
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacuag                                         4107


<210>  218
<211>  4101
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase coding sequence encoding SEQ ID NO: 216 
       using minimal uridine codons as listed in Table 3 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  218
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga c                                               4101


<210>  219
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of dCas9 (without NLS)

<400>  219

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp Ala 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             


Ser 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             


<210>  220
<211>  4107
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 mRNA ORF encoding SEQ ID NO: 219 using minimal 
       uridine codons as listed in Table 3, with start and stop codons

<400>  220
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgacgca     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacuag                                         4107


<210>  221
<211>  4113
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 coding sequence encoding SEQ ID NO: 219 using 
       minimal uridine codons as listed in Table 3 (no start or stop 
       codons; suitable for inclusion in fusion protein coding sequence)

<400>  221
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgacgcaauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agc                                  4113


<210>  222
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 with two nuclear 
       localization signals (2xNLS) as the C-terminal amino acids

<400>  222

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    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             


Ser 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             


Gly Ser  Gly Ser Pro Lys Lys  Lys Arg Lys Val Asp  Gly Ser Pro 
    1370                 1375                 1380             


Lys Lys  Lys Arg Lys Val Asp  Ser Gly 
    1385                 1390         


<210>  223
<211>  4233
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF encoding SEQ ID NO: 222 using minimal 
       uridine codons, with start and stop codons

<400>  223
auggacaaga aguacagcau cggacuggac aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucccg     4140

aagaagaaga gaaaggucgg aagcggaagc ccgaagaaga agagaaaggu cgacggaagc     4200

ccgaagaaga agagaaaggu cgacagcgga uag                                  4233


<210>  224
<211>  4227
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 coding sequence encoding SEQ ID NO: 222 using 
       minimal uridine codons (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  224
gacaagaagu acagcaucgg acuggacauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa ggucccgaag     4140

aagaagagaa aggucggaag cggaagcccg aagaagaaga gaaaggucga cggaagcccg     4200

aagaagaaga gaaaggucga cagcgga                                         4227


<210>  225
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 nickase with two nuclear 
       localization signals as the C-terminal amino acids

<400>  225

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    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             


Ser 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             


Gly Ser  Gly Ser Pro Lys Lys  Lys Arg Lys Val Asp  Gly Ser Pro 
    1370                 1375                 1380             


Lys Lys  Lys Arg Lys Val Asp  Ser Gly 
    1385                 1390         


<210>  226
<211>  4179
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase mRNA ORF encoding SEQ ID NO: 25 using 
       minimal uridine codons as listed in Table 3, with start and stop 
       codons

<400>  226
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggaagc ggaagcccga agaagaagag aaaggucgac     4140

ggaagcccga agaagaagag aaaggucgac agcggauag                            4179


<210>  227
<211>  4173
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase coding sequence encoding SEQ ID NO: 25 
       using minimal uridine codons (no start or stop codons; suitable 
       for inclusion in fusion protein coding sequence)

<400>  227
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaagcgga agcccgaaga agaagagaaa ggucgacgga     4140

agcccgaaga agaagagaaa ggucgacagc gga                                  4173


<210>  228
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of dCas9 with two nuclear 
       localization signals as the C-terminal  amino acids

<400>  228

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


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


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


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


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


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


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


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp Ala 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             


Ser 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             


Gly Ser  Gly Ser Pro Lys Lys  Lys Arg Lys Val Asp  Gly Ser Pro 
    1370                 1375                 1380             


Lys Lys  Lys Arg Lys Val Asp  Ser Gly 
    1385                 1390         


<210>  229
<211>  4179
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 mRNA ORF encoding SEQ ID NO: 228 using minimal 
       uridine codons, with start and stop codons

<400>  229
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgacgca     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggaagc ggaagcccga agaagaagag aaaggucgac     4140

ggaagcccga agaagaagag aaaggucgac agcggauag                            4179


<210>  230
<211>  4173
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 coding sequence encoding SEQ ID NO: 228 using 
       minimal uridine codons (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  230
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgacgcaauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaagcgga agcccgaaga agaagagaaa ggucgacgga     4140

agcccgaaga agaagagaaa ggucgacagc gga                                  4173


<210>  231
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: T7 Promoter

<400>  231
taatacgact cactata                                                      17


<210>  232
<211>  50
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(50)
<223>  Human beta-globin 5 UTR

<400>  232
acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc                  50


<210>  233
<211>  132
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(132)
<223>  Human beta-globin 3 UTR

<400>  233
gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac       60

taaactgggg gatattatga agggccttga gcatctggat tctgcctaat aaaaaacatt      120

tattttcatt gc                                                          132


<210>  234
<211>  66
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(66)
<223>  Human alpha-globin 5 UTR

<400>  234
cataaaccct ggcgcgctcg cggcccggca ctcttctggt ccccacagac tcagagagaa       60

cccacc                                                                  66


<210>  235
<211>  110
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(110)
<223>  Human alpha-globin 3 UTR

<400>  235
gctggagcct cggtggccat gcttcttgcc ccttgggcct ccccccagcc cctcctcccc       60

ttcctgcacc cgtacccccg tggtctttga ataaagtctg agtgggcggc                 110


<210>  236
<211>  29
<212>  DNA
<213>  Xenopus laevis


<220>
<221>  misc_feature
<222>  (1)..(29)
<223>  Xenopus laevis beta-globin 5 UTR

<400>  236
aagctcagaa taaacgctca actttggcc                                         29


<210>  237
<211>  130
<212>  DNA
<213>  Xenopus laevis


<220>
<221>  misc_feature
<222>  (1)..(130)
<223>  Xenopus laevis beta-globin 3 UTR

<400>  237
accagcctca agaacacccg aatggagtct ctaagctaca taataccaac ttacacttta       60

caaaatgttg tcccccaaaa tgtagccatt cgtatctgct cctaataaaa agaaagtttc      120

ttcacattct                                                             130


<210>  238
<211>  27
<212>  DNA
<213>  Bos taurus


<220>
<221>  misc_feature
<222>  (1)..(27)
<223>  Bovine Growth Hormone 5 UTR

<400>  238
cagggtcctg tggacagctc accagct                                           27


<210>  239
<211>  102
<212>  DNA
<213>  Bos taurus


<220>
<221>  misc_feature
<222>  (1)..(102)
<223>  Bovine Growth Hormone 3 UTR

<400>  239
ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac       60

tcccactgtc ctttcctaat aaaatgagga aattgcatcg ca                         102


<210>  240
<211>  93
<212>  DNA
<213>  Mus musculus


<220>
<221>  misc_feature
<222>  (1)..(93)
<223>  Mus musculus hemoglobin alpha, adult chain 1 (Hba-a1), 3UTR

<400>  240
gctgccttct gcggggcttg ccttctggcc atgcccttct tctctccctt gcacctgtac       60

ctcttggtct ttgaataaag cctgagtagg aag                                    93


<210>  241
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: HSD17B4 5 UTR

<400>  241
tcccgcagtc ggcgtccagc ggctctgctt gttcgtgtgt gtgtcgttgc aggccttatt       60

c                                                                       61


<210>  242
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G282 single guide RNA targeting the mouse TTR gene


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  242
uuacagccac gucuacagca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  243

<400>  243
000

<210>  244
<211>  4405
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 204, and 3 UTR of ALB

<400>  244
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg      120

atgggcagtc atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa      180

cacagacaga cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga      240

aacagcagaa gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa      300

cagaatctgc tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt      360

cttccacaga ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc      420

gatcttcgga aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca      480

cctgagaaag aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc      540

actggcacac atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga      600

caacagcgac gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga      660

agaaaacccg atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag      720

caagagcaga agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact      780

gttcggaaac ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga      840

cctggcagaa gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa      900

cctgctggca cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag      960

cgacgcaatc ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct     1020

gagcgcaagc atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc     1080

actggtcaga cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa     1140

cggatacgca ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa     1200

gccgatcctg gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga     1260

cctgctgaga aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg     1320

agaactgcac gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag     1380

agaaaagatc gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag     1440

aggaaacagc agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa     1500

cttcgaagaa gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa     1560

cttcgacaag aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata     1620

cttcacagtc tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc     1680

ggcattcctg agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag     1740

aaaggtcaca gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag     1800

cgtcgaaatc agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct     1860

gctgaagatc atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga     1920

agacatcgtc ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa     1980

gacatacgca cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac     2040

aggatgggga agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa     2100

gacaatcctg gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat     2160

ccacgacgac agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg     2220

agacagcctg cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat     2280

cctgcagaca gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga     2340

aaacatcgtc atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag     2400

cagagaaaga atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa     2460

ggaacacccg gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca     2520

gaacggaaga gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga     2580

cgtcgaccac atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct     2640

gacaagaagc gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa     2700

gaagatgaag aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt     2760

cgacaacctg acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat     2820

caagagacag ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag     2880

cagaatgaac acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac     2940

actgaagagc aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga     3000

aatcaacaac taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact     3060

gatcaagaag tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga     3120

cgtcagaaag atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt     3180

cttctacagc aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat     3240

cagaaagaga ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg     3300

aagagacttc gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa     3360

gacagaagtc cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga     3420

caagctgatc gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc     3480

gacagtcgca tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct     3540

gaagagcgtc aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa     3600

cccgatcgac ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa     3660

gctgccgaag tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc     3720

aggagaactg cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta     3780

cctggcaagc cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct     3840

gttcgtcgaa cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag     3900

caagagagtc atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca     3960

cagagacaag ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa     4020

cctgggagca ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac     4080

aagcacaaag gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga     4140

aacaagaatc gacctgagcc agctgggagg agacggagga ggaagcccga agaagaagag     4200

aaaggtctag ctagccatca catttaaaag catctcagcc taccatgaga ataagagaaa     4260

gaaaatgaag atcaatagct tattcatctc tttttctttt tcgttggtgt aaagccaaca     4320

ccctgtctaa aaaacataaa tttctttaat cattttgcct cttttctctg tgcttcaatt     4380

aataaaaaat ggaaagaacc tcgag                                           4405


<210>  245
<211>  4188
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Alternative Cas9 ORF with 19.36% U content

<400>  245
atggataaga agtactcgat cgggctggat atcggaacta attccgtggg ttgggcagtg       60

atcacggatg aatacaaagt gccgtccaag aagttcaagg tcctggggaa caccgataga      120

cacagcatca agaagaatct catcggagcc ctgctgtttg actccggcga aaccgcagaa      180

gcgacccggc tcaaacgtac cgcgaggcga cgctacaccc ggcggaagaa tcgcatctgc      240

tatctgcaag aaatcttttc gaacgaaatg gcaaaggtgg acgacagctt cttccaccgc      300

ctggaagaat ctttcctggt ggaggaggac aagaagcatg aacggcatcc tatctttgga      360

aacatcgtgg acgaagtggc gtaccacgaa aagtacccga ccatctacca tctgcggaag      420

aagttggttg actcaactga caaggccgac ctcagattga tctacttggc cctcgcccat      480

atgatcaaat tccgcggaca cttcctgatc gaaggcgatc tgaaccctga taactccgac      540

gtggataagc tgttcattca actggtgcag acctacaacc aactgttcga agaaaaccca      600

atcaatgcca gcggcgtcga tgccaaggcc atcctgtccg cccggctgtc gaagtcgcgg      660

cgcctcgaaa acctgatcgc acagctgccg ggagagaaga agaacggact tttcggcaac      720

ttgatcgctc tctcactggg actcactccc aatttcaagt ccaattttga cctggccgag      780

gacgcgaagc tgcaactctc aaaggacacc tacgacgacg acttggacaa tttgctggca      840

caaattggcg atcagtacgc ggatctgttc cttgccgcta agaacctttc ggacgcaatc      900

ttgctgtccg atatcctgcg cgtgaacacc gaaataacca aagcgccgct tagcgcctcg      960

atgattaagc ggtacgacga gcatcaccag gatctcacgc tgctcaaagc gctcgtgaga     1020

cagcaactgc ctgaaaagta caaggagatt ttcttcgacc agtccaagaa tgggtacgca     1080

gggtacatcg atggaggcgc cagccaggaa gagttctata agttcatcaa gccaatcctg     1140

gaaaagatgg acggaaccga agaactgctg gtcaagctga acagggagga tctgctccgc     1200

aaacagagaa cctttgacaa cggaagcatt ccacaccaga tccatctggg tgagctgcac     1260

gccatcttgc ggcgccagga ggacttttac ccattcctca aggacaaccg ggaaaagatc     1320

gagaaaattc tgacgttccg catcccgtat tacgtgggcc cactggcgcg cggcaattcg     1380

cgcttcgcgt ggatgactag aaaatcagag gaaaccatca ctccttggaa tttcgaggaa     1440

gttgtggata agggagcttc ggcacaatcc ttcatcgaac gaatgaccaa cttcgacaag     1500

aatctcccaa acgagaaggt gcttcctaag cacagcctcc tttacgaata cttcactgtc     1560

tacaacgaac tgactaaagt gaaatacgtt actgaaggaa tgaggaagcc ggcctttctg     1620

agcggagaac agaagaaagc gattgtcgat ctgctgttca agaccaaccg caaggtgacc     1680

gtcaagcagc ttaaagagga ctacttcaag aagatcgagt gtttcgactc agtggaaatc     1740

agcggagtgg aggacagatt caacgcttcg ctgggaacct atcatgatct cctgaagatc     1800

atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctgga agatatcgtc     1860

ctgaccttga cccttttcga ggatcgcgag atgatcgagg agaggcttaa gacctacgct     1920

catctcttcg acgataaggt catgaaacaa ctcaagcgcc gccggtacac tggttggggc     1980

cgcctctccc gcaagctgat caacggtatt cgcgataaac agagcggtaa aactatcctg     2040

gatttcctca aatcggatgg cttcgctaat cgtaacttca tgcagttgat ccacgacgac     2100

agcctgacct ttaaggagga catccagaaa gcacaagtga gcggacaggg agactcactc     2160

catgaacaca tcgcgaatct ggccggttcg ccggcgatta agaagggaat cctgcaaact     2220

gtgaaggtgg tggacgagct ggtgaaggtc atgggacggc acaaaccgga gaatatcgtg     2280

attgaaatgg cccgagaaaa ccagactacc cagaagggcc agaagaactc ccgcgaaagg     2340

atgaagcgga tcgaagaagg aatcaaggag ctgggcagcc agatcctgaa agagcacccg     2400

gtggaaaaca cgcagctgca gaacgagaag ctctacctgt actatttgca aaatggacgg     2460

gacatgtacg tggaccaaga gctggacatc aatcggttgt ctgattacga cgtggaccac     2520

atcgttccac agtcctttct gaaggatgac tccatcgata acaaggtgtt gactcgcagc     2580

gacaagaaca gagggaagtc agataatgtg ccatcggagg aggtcgtgaa gaagatgaag     2640

aattactggc ggcagctcct gaatgcgaag ctgattaccc agagaaagtt tgacaatctc     2700

actaaagccg agcgcggcgg actctcagag ctggataagg ctggattcat caaacggcag     2760

ctggtcgaga ctcggcagat taccaagcac gtggcgcaga tcctggactc ccgcatgaac     2820

actaaatacg acgagaacga taagctcatc cgggaagtga aggtgattac cctgaaaagc     2880

aaacttgtgt cggactttcg gaaggacttt cagttttaca aagtgagaga aatcaacaac     2940

taccatcacg cgcatgacgc atacctcaac gctgtggtcg gcaccgccct gatcaagaag     3000

taccctaaac ttgaatcgga gtttgtgtac ggagactaca aggtctacga cgtgaggaag     3060

atgatagcca agtccgaaca ggaaatcggg aaagcaactg cgaaatactt cttttactca     3120

aacatcatga acttcttcaa gactgaaatt acgctggcca atggagaaat caggaagagg     3180

ccactgatcg aaactaacgg agaaacgggc gaaatcgtgt gggacaaggg cagggacttc     3240

gcaactgttc gcaaagtgct ctctatgccg caagtcaata ttgtgaagaa aaccgaagtg     3300

caaaccggcg gattttcaaa ggaatcgatc ctcccaaaga gaaatagcga caagctcatt     3360

gcacgcaaga aagactggga cccgaagaag tacggaggat tcgattcgcc gactgtcgca     3420

tactccgtcc tcgtggtggc caaggtggag aagggaaaga gcaagaagct caaatccgtc     3480

aaagagctgc tggggattac catcatggaa cgatcctcgt tcgagaagaa cccgattgat     3540

ttcctggagg cgaagggtta caaggaggtg aagaaggatc tgatcatcaa actgcccaag     3600

tactcactgt tcgaactgga aaatggtcgg aagcgcatgc tggcttcggc cggagaactc     3660

cagaaaggaa atgagctggc cttgcctagc aagtacgtca acttcctcta tcttgcttcg     3720

cactacgaga aactcaaagg gtcaccggaa gataacgaac agaagcagct tttcgtggag     3780

cagcacaagc attatctgga tgaaatcatc gaacaaatct ccgagttttc aaagcgcgtg     3840

atcctcgccg acgccaacct cgacaaagtc ctgtcggcct acaataagca tagagataag     3900

ccgatcagag aacaggccga gaacattatc cacttgttca ccctgactaa cctgggagct     3960

ccagccgcct tcaagtactt cgatactact atcgaccgca aaagatacac gtccaccaag     4020

gaagttctgg acgcgaccct gatccaccaa agcatcactg gactctacga aactaggatc     4080

gatctgtcgc agctgggtgg cgatggtggc ggtggatcct acccatacga cgtgcctgac     4140

tacgcctccg gaggtggtgg ccccaagaag aaacggaagg tgtgatag                  4188


<210>  246
<211>  4459
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 245, Kozak sequence, and 3 UTR of ALB

<400>  246
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatct gccaccatgg ataagaagta ctcgatcggg ctggatatcg gaactaattc      120

cgtgggttgg gcagtgatca cggatgaata caaagtgccg tccaagaagt tcaaggtcct      180

ggggaacacc gatagacaca gcatcaagaa gaatctcatc ggagccctgc tgtttgactc      240

cggcgaaacc gcagaagcga cccggctcaa acgtaccgcg aggcgacgct acacccggcg      300

gaagaatcgc atctgctatc tgcaagaaat cttttcgaac gaaatggcaa aggtggacga      360

cagcttcttc caccgcctgg aagaatcttt cctggtggag gaggacaaga agcatgaacg      420

gcatcctatc tttggaaaca tcgtggacga agtggcgtac cacgaaaagt acccgaccat      480

ctaccatctg cggaagaagt tggttgactc aactgacaag gccgacctca gattgatcta      540

cttggccctc gcccatatga tcaaattccg cggacacttc ctgatcgaag gcgatctgaa      600

ccctgataac tccgacgtgg ataagctgtt cattcaactg gtgcagacct acaaccaact      660

gttcgaagaa aacccaatca atgccagcgg cgtcgatgcc aaggccatcc tgtccgcccg      720

gctgtcgaag tcgcggcgcc tcgaaaacct gatcgcacag ctgccgggag agaagaagaa      780

cggacttttc ggcaacttga tcgctctctc actgggactc actcccaatt tcaagtccaa      840

ttttgacctg gccgaggacg cgaagctgca actctcaaag gacacctacg acgacgactt      900

ggacaatttg ctggcacaaa ttggcgatca gtacgcggat ctgttccttg ccgctaagaa      960

cctttcggac gcaatcttgc tgtccgatat cctgcgcgtg aacaccgaaa taaccaaagc     1020

gccgcttagc gcctcgatga ttaagcggta cgacgagcat caccaggatc tcacgctgct     1080

caaagcgctc gtgagacagc aactgcctga aaagtacaag gagattttct tcgaccagtc     1140

caagaatggg tacgcagggt acatcgatgg aggcgccagc caggaagagt tctataagtt     1200

catcaagcca atcctggaaa agatggacgg aaccgaagaa ctgctggtca agctgaacag     1260

ggaggatctg ctccgcaaac agagaacctt tgacaacgga agcattccac accagatcca     1320

tctgggtgag ctgcacgcca tcttgcggcg ccaggaggac ttttacccat tcctcaagga     1380

caaccgggaa aagatcgaga aaattctgac gttccgcatc ccgtattacg tgggcccact     1440

ggcgcgcggc aattcgcgct tcgcgtggat gactagaaaa tcagaggaaa ccatcactcc     1500

ttggaatttc gaggaagttg tggataaggg agcttcggca caatccttca tcgaacgaat     1560

gaccaacttc gacaagaatc tcccaaacga gaaggtgctt cctaagcaca gcctccttta     1620

cgaatacttc actgtctaca acgaactgac taaagtgaaa tacgttactg aaggaatgag     1680

gaagccggcc tttctgagcg gagaacagaa gaaagcgatt gtcgatctgc tgttcaagac     1740

caaccgcaag gtgaccgtca agcagcttaa agaggactac ttcaagaaga tcgagtgttt     1800

cgactcagtg gaaatcagcg gagtggagga cagattcaac gcttcgctgg gaacctatca     1860

tgatctcctg aagatcatca aggacaagga cttccttgac aacgaggaga acgaggacat     1920

cctggaagat atcgtcctga ccttgaccct tttcgaggat cgcgagatga tcgaggagag     1980

gcttaagacc tacgctcatc tcttcgacga taaggtcatg aaacaactca agcgccgccg     2040

gtacactggt tggggccgcc tctcccgcaa gctgatcaac ggtattcgcg ataaacagag     2100

cggtaaaact atcctggatt tcctcaaatc ggatggcttc gctaatcgta acttcatgca     2160

gttgatccac gacgacagcc tgacctttaa ggaggacatc cagaaagcac aagtgagcgg     2220

acagggagac tcactccatg aacacatcgc gaatctggcc ggttcgccgg cgattaagaa     2280

gggaatcctg caaactgtga aggtggtgga cgagctggtg aaggtcatgg gacggcacaa     2340

accggagaat atcgtgattg aaatggcccg agaaaaccag actacccaga agggccagaa     2400

gaactcccgc gaaaggatga agcggatcga agaaggaatc aaggagctgg gcagccagat     2460

cctgaaagag cacccggtgg aaaacacgca gctgcagaac gagaagctct acctgtacta     2520

tttgcaaaat ggacgggaca tgtacgtgga ccaagagctg gacatcaatc ggttgtctga     2580

ttacgacgtg gaccacatcg ttccacagtc ctttctgaag gatgactcca tcgataacaa     2640

ggtgttgact cgcagcgaca agaacagagg gaagtcagat aatgtgccat cggaggaggt     2700

cgtgaagaag atgaagaatt actggcggca gctcctgaat gcgaagctga ttacccagag     2760

aaagtttgac aatctcacta aagccgagcg cggcggactc tcagagctgg ataaggctgg     2820

attcatcaaa cggcagctgg tcgagactcg gcagattacc aagcacgtgg cgcagatcct     2880

ggactcccgc atgaacacta aatacgacga gaacgataag ctcatccggg aagtgaaggt     2940

gattaccctg aaaagcaaac ttgtgtcgga ctttcggaag gactttcagt tttacaaagt     3000

gagagaaatc aacaactacc atcacgcgca tgacgcatac ctcaacgctg tggtcggcac     3060

cgccctgatc aagaagtacc ctaaacttga atcggagttt gtgtacggag actacaaggt     3120

ctacgacgtg aggaagatga tagccaagtc cgaacaggaa atcgggaaag caactgcgaa     3180

atacttcttt tactcaaaca tcatgaactt cttcaagact gaaattacgc tggccaatgg     3240

agaaatcagg aagaggccac tgatcgaaac taacggagaa acgggcgaaa tcgtgtggga     3300

caagggcagg gacttcgcaa ctgttcgcaa agtgctctct atgccgcaag tcaatattgt     3360

gaagaaaacc gaagtgcaaa ccggcggatt ttcaaaggaa tcgatcctcc caaagagaaa     3420

tagcgacaag ctcattgcac gcaagaaaga ctgggacccg aagaagtacg gaggattcga     3480

ttcgccgact gtcgcatact ccgtcctcgt ggtggccaag gtggagaagg gaaagagcaa     3540

gaagctcaaa tccgtcaaag agctgctggg gattaccatc atggaacgat cctcgttcga     3600

gaagaacccg attgatttcc tggaggcgaa gggttacaag gaggtgaaga aggatctgat     3660

catcaaactg cccaagtact cactgttcga actggaaaat ggtcggaagc gcatgctggc     3720

ttcggccgga gaactccaga aaggaaatga gctggccttg cctagcaagt acgtcaactt     3780

cctctatctt gcttcgcact acgagaaact caaagggtca ccggaagata acgaacagaa     3840

gcagcttttc gtggagcagc acaagcatta tctggatgaa atcatcgaac aaatctccga     3900

gttttcaaag cgcgtgatcc tcgccgacgc caacctcgac aaagtcctgt cggcctacaa     3960

taagcataga gataagccga tcagagaaca ggccgagaac attatccact tgttcaccct     4020

gactaacctg ggagctccag ccgccttcaa gtacttcgat actactatcg accgcaaaag     4080

atacacgtcc accaaggaag ttctggacgc gaccctgatc caccaaagca tcactggact     4140

ctacgaaact aggatcgatc tgtcgcagct gggtggcgat ggtggcggtg gatcctaccc     4200

atacgacgtg cctgactacg cctccggagg tggtggcccc aagaagaaac ggaaggtgtg     4260

atagctagcc atcacattta aaagcatctc agcctaccat gagaataaga gaaagaaaat     4320

gaagatcaat agcttattca tctctttttc tttttcgttg gtgtaaagcc aacaccctgt     4380

ctaaaaaaca taaatttctt taatcatttt gcctcttttc tctgtgcttc aattaataaa     4440

aaatggaaag aacctcgag                                                  4459


<210>  247
<211>  4453
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 245, and 3 UTR of ALB

<400>  247
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatct atggataaga agtactcgat cgggctggat atcggaacta attccgtggg      120

ttgggcagtg atcacggatg aatacaaagt gccgtccaag aagttcaagg tcctggggaa      180

caccgataga cacagcatca agaagaatct catcggagcc ctgctgtttg actccggcga      240

aaccgcagaa gcgacccggc tcaaacgtac cgcgaggcga cgctacaccc ggcggaagaa      300

tcgcatctgc tatctgcaag aaatcttttc gaacgaaatg gcaaaggtgg acgacagctt      360

cttccaccgc ctggaagaat ctttcctggt ggaggaggac aagaagcatg aacggcatcc      420

tatctttgga aacatcgtgg acgaagtggc gtaccacgaa aagtacccga ccatctacca      480

tctgcggaag aagttggttg actcaactga caaggccgac ctcagattga tctacttggc      540

cctcgcccat atgatcaaat tccgcggaca cttcctgatc gaaggcgatc tgaaccctga      600

taactccgac gtggataagc tgttcattca actggtgcag acctacaacc aactgttcga      660

agaaaaccca atcaatgcca gcggcgtcga tgccaaggcc atcctgtccg cccggctgtc      720

gaagtcgcgg cgcctcgaaa acctgatcgc acagctgccg ggagagaaga agaacggact      780

tttcggcaac ttgatcgctc tctcactggg actcactccc aatttcaagt ccaattttga      840

cctggccgag gacgcgaagc tgcaactctc aaaggacacc tacgacgacg acttggacaa      900

tttgctggca caaattggcg atcagtacgc ggatctgttc cttgccgcta agaacctttc      960

ggacgcaatc ttgctgtccg atatcctgcg cgtgaacacc gaaataacca aagcgccgct     1020

tagcgcctcg atgattaagc ggtacgacga gcatcaccag gatctcacgc tgctcaaagc     1080

gctcgtgaga cagcaactgc ctgaaaagta caaggagatt ttcttcgacc agtccaagaa     1140

tgggtacgca gggtacatcg atggaggcgc cagccaggaa gagttctata agttcatcaa     1200

gccaatcctg gaaaagatgg acggaaccga agaactgctg gtcaagctga acagggagga     1260

tctgctccgc aaacagagaa cctttgacaa cggaagcatt ccacaccaga tccatctggg     1320

tgagctgcac gccatcttgc ggcgccagga ggacttttac ccattcctca aggacaaccg     1380

ggaaaagatc gagaaaattc tgacgttccg catcccgtat tacgtgggcc cactggcgcg     1440

cggcaattcg cgcttcgcgt ggatgactag aaaatcagag gaaaccatca ctccttggaa     1500

tttcgaggaa gttgtggata agggagcttc ggcacaatcc ttcatcgaac gaatgaccaa     1560

cttcgacaag aatctcccaa acgagaaggt gcttcctaag cacagcctcc tttacgaata     1620

cttcactgtc tacaacgaac tgactaaagt gaaatacgtt actgaaggaa tgaggaagcc     1680

ggcctttctg agcggagaac agaagaaagc gattgtcgat ctgctgttca agaccaaccg     1740

caaggtgacc gtcaagcagc ttaaagagga ctacttcaag aagatcgagt gtttcgactc     1800

agtggaaatc agcggagtgg aggacagatt caacgcttcg ctgggaacct atcatgatct     1860

cctgaagatc atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctgga     1920

agatatcgtc ctgaccttga cccttttcga ggatcgcgag atgatcgagg agaggcttaa     1980

gacctacgct catctcttcg acgataaggt catgaaacaa ctcaagcgcc gccggtacac     2040

tggttggggc cgcctctccc gcaagctgat caacggtatt cgcgataaac agagcggtaa     2100

aactatcctg gatttcctca aatcggatgg cttcgctaat cgtaacttca tgcagttgat     2160

ccacgacgac agcctgacct ttaaggagga catccagaaa gcacaagtga gcggacaggg     2220

agactcactc catgaacaca tcgcgaatct ggccggttcg ccggcgatta agaagggaat     2280

cctgcaaact gtgaaggtgg tggacgagct ggtgaaggtc atgggacggc acaaaccgga     2340

gaatatcgtg attgaaatgg cccgagaaaa ccagactacc cagaagggcc agaagaactc     2400

ccgcgaaagg atgaagcgga tcgaagaagg aatcaaggag ctgggcagcc agatcctgaa     2460

agagcacccg gtggaaaaca cgcagctgca gaacgagaag ctctacctgt actatttgca     2520

aaatggacgg gacatgtacg tggaccaaga gctggacatc aatcggttgt ctgattacga     2580

cgtggaccac atcgttccac agtcctttct gaaggatgac tccatcgata acaaggtgtt     2640

gactcgcagc gacaagaaca gagggaagtc agataatgtg ccatcggagg aggtcgtgaa     2700

gaagatgaag aattactggc ggcagctcct gaatgcgaag ctgattaccc agagaaagtt     2760

tgacaatctc actaaagccg agcgcggcgg actctcagag ctggataagg ctggattcat     2820

caaacggcag ctggtcgaga ctcggcagat taccaagcac gtggcgcaga tcctggactc     2880

ccgcatgaac actaaatacg acgagaacga taagctcatc cgggaagtga aggtgattac     2940

cctgaaaagc aaacttgtgt cggactttcg gaaggacttt cagttttaca aagtgagaga     3000

aatcaacaac taccatcacg cgcatgacgc atacctcaac gctgtggtcg gcaccgccct     3060

gatcaagaag taccctaaac ttgaatcgga gtttgtgtac ggagactaca aggtctacga     3120

cgtgaggaag atgatagcca agtccgaaca ggaaatcggg aaagcaactg cgaaatactt     3180

cttttactca aacatcatga acttcttcaa gactgaaatt acgctggcca atggagaaat     3240

caggaagagg ccactgatcg aaactaacgg agaaacgggc gaaatcgtgt gggacaaggg     3300

cagggacttc gcaactgttc gcaaagtgct ctctatgccg caagtcaata ttgtgaagaa     3360

aaccgaagtg caaaccggcg gattttcaaa ggaatcgatc ctcccaaaga gaaatagcga     3420

caagctcatt gcacgcaaga aagactggga cccgaagaag tacggaggat tcgattcgcc     3480

gactgtcgca tactccgtcc tcgtggtggc caaggtggag aagggaaaga gcaagaagct     3540

caaatccgtc aaagagctgc tggggattac catcatggaa cgatcctcgt tcgagaagaa     3600

cccgattgat ttcctggagg cgaagggtta caaggaggtg aagaaggatc tgatcatcaa     3660

actgcccaag tactcactgt tcgaactgga aaatggtcgg aagcgcatgc tggcttcggc     3720

cggagaactc cagaaaggaa atgagctggc cttgcctagc aagtacgtca acttcctcta     3780

tcttgcttcg cactacgaga aactcaaagg gtcaccggaa gataacgaac agaagcagct     3840

tttcgtggag cagcacaagc attatctgga tgaaatcatc gaacaaatct ccgagttttc     3900

aaagcgcgtg atcctcgccg acgccaacct cgacaaagtc ctgtcggcct acaataagca     3960

tagagataag ccgatcagag aacaggccga gaacattatc cacttgttca ccctgactaa     4020

cctgggagct ccagccgcct tcaagtactt cgatactact atcgaccgca aaagatacac     4080

gtccaccaag gaagttctgg acgcgaccct gatccaccaa agcatcactg gactctacga     4140

aactaggatc gatctgtcgc agctgggtgg cgatggtggc ggtggatcct acccatacga     4200

cgtgcctgac tacgcctccg gaggtggtgg ccccaagaag aaacggaagg tgtgatagct     4260

agccatcaca tttaaaagca tctcagccta ccatgagaat aagagaaaga aaatgaagat     4320

caatagctta ttcatctctt tttctttttc gttggtgtaa agccaacacc ctgtctaaaa     4380

aacataaatt tctttaatca ttttgcctct tttctctgtg cttcaattaa taaaaaatgg     4440

aaagaacctc gag                                                        4453


<210>  248

<400>  248
000

<210>  249
<211>  4409
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript comprising Kozak sequence with Cas9 
       ORF using codons with generally high expression in humans

<400>  249
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgc ctaagaaaaa gcggaaggtc gacggggata agaagtactc      120

aatcgggctg gatatcggaa ctaattccgt gggttgggca gtgatcacgg atgaatacaa      180

agtgccgtcc aagaagttca aggtcctggg gaacaccgat agacacagca tcaagaaaaa      240

tctcatcgga gccctgctgt ttgactccgg cgaaaccgca gaagcgaccc ggctcaaacg      300

taccgcgagg cgacgctaca cccggcggaa gaatcgcatc tgctatctgc aagagatctt      360

ttcgaacgaa atggcaaagg tcgacgacag cttcttccac cgcctggaag aatctttcct      420

ggtggaggag gacaagaagc atgaacggca tcctatcttt ggaaacatcg tcgacgaagt      480

ggcgtaccac gaaaagtacc cgaccatcta ccatctgcgg aagaagttgg ttgactcaac      540

tgacaaggcc gacctcagat tgatctactt ggccctcgcc catatgatca aattccgcgg      600

acacttcctg atcgaaggcg atctgaaccc tgataactcc gacgtggata agcttttcat      660

tcaactggtg cagacctaca accaactgtt cgaagaaaac ccaatcaatg ctagcggcgt      720

cgatgccaag gccatcctgt ccgcccggct gtcgaagtcg cggcgcctcg aaaacctgat      780

cgcacagctg ccgggagaga aaaagaacgg acttttcggc aacttgatcg ctctctcact      840

gggactcact cccaatttca agtccaattt tgacctggcc gaggacgcga agctgcaact      900

ctcaaaggac acctacgacg acgacttgga caatttgctg gcacaaattg gcgatcagta      960

cgcggatctg ttccttgccg ctaagaacct ttcggacgca atcttgctgt ccgatatcct     1020

gcgcgtgaac accgaaataa ccaaagcgcc gcttagcgcc tcgatgatta agcggtacga     1080

cgagcatcac caggatctca cgctgctcaa agcgctcgtg agacagcaac tgcctgaaaa     1140

gtacaaggag atcttcttcg accagtccaa gaatgggtac gcagggtaca tcgatggagg     1200

cgctagccag gaagagttct ataagttcat caagccaatc ctggaaaaga tggacggaac     1260

cgaagaactg ctggtcaagc tgaacaggga ggatctgctc cggaaacaga gaacctttga     1320

caacggatcc attccccacc agatccatct gggtgagctg cacgccatct tgcggcgcca     1380

ggaggacttt tacccattcc tcaaggacaa ccgggaaaag atcgagaaaa ttctgacgtt     1440

ccgcatcccg tattacgtgg gcccactggc gcgcggcaat tcgcgcttcg cgtggatgac     1500

tagaaaatca gaggaaacca tcactccttg gaatttcgag gaagttgtgg ataagggagc     1560

ttcggcacaa agcttcatcg aacgaatgac caacttcgac aagaatctcc caaacgagaa     1620

ggtgcttcct aagcacagcc tcctttacga atacttcact gtctacaacg aactgactaa     1680

agtgaaatac gttactgaag gaatgaggaa gccggccttt ctgtccggag aacagaagaa     1740

agcaattgtc gatctgctgt tcaagaccaa ccgcaaggtg accgtcaagc agcttaaaga     1800

ggactacttc aagaagatcg agtgtttcga ctcagtggaa atcagcgggg tggaggacag     1860

attcaacgct tcgctgggaa cctatcatga tctcctgaag atcatcaagg acaaggactt     1920

ccttgacaac gaggagaacg aggacatcct ggaagatatc gtcctgacct tgaccctttt     1980

cgaggatcgc gagatgatcg aggagaggct taagacctac gctcatctct tcgacgataa     2040

ggtcatgaaa caactcaagc gccgccggta cactggttgg ggccgcctct cccgcaagct     2100

gatcaacggt attcgcgata aacagagcgg taaaactatc ctggatttcc tcaaatcgga     2160

tggcttcgct aatcgtaact tcatgcaatt gatccacgac gacagcctga cctttaagga     2220

ggacatccaa aaagcacaag tgtccggaca gggagactca ctccatgaac acatcgcgaa     2280

tctggccggt tcgccggcga ttaagaaggg aattctgcaa actgtgaagg tggtcgacga     2340

gctggtgaag gtcatgggac ggcacaaacc ggagaatatc gtgattgaaa tggcccgaga     2400

aaaccagact acccagaagg gccagaaaaa ctcccgcgaa aggatgaagc ggatcgaaga     2460

aggaatcaag gagctgggca gccagatcct gaaagagcac ccggtggaaa acacgcagct     2520

gcagaacgag aagctctacc tgtactattt gcaaaatgga cgggacatgt acgtggacca     2580

agagctggac atcaatcggt tgtctgatta cgacgtggac cacatcgttc cacagtcctt     2640

tctgaaggat gactcgatcg ataacaaggt gttgactcgc agcgacaaga acagagggaa     2700

gtcagataat gtgccatcgg aggaggtcgt gaagaagatg aagaattact ggcggcagct     2760

cctgaatgcg aagctgatta cccagagaaa gtttgacaat ctcactaaag ccgagcgcgg     2820

cggactctca gagctggata aggctggatt catcaaacgg cagctggtcg agactcggca     2880

gattaccaag cacgtggcgc agatcttgga ctcccgcatg aacactaaat acgacgagaa     2940

cgataagctc atccgggaag tgaaggtgat taccctgaaa agcaaacttg tgtcggactt     3000

tcggaaggac tttcagtttt acaaagtgag agaaatcaac aactaccatc acgcgcatga     3060

cgcatacctc aacgctgtgg tcggtaccgc cctgatcaaa aagtacccta aacttgaatc     3120

ggagtttgtg tacggagact acaaggtcta cgacgtgagg aagatgatag ccaagtccga     3180

acaggaaatc gggaaagcaa ctgcgaaata cttcttttac tcaaacatca tgaacttttt     3240

caagactgaa attacgctgg ccaatggaga aatcaggaag aggccactga tcgaaactaa     3300

cggagaaacg ggcgaaatcg tgtgggacaa gggcagggac ttcgcaactg ttcgcaaagt     3360

gctctctatg ccgcaagtca atattgtgaa gaaaaccgaa gtgcaaaccg gcggattttc     3420

aaaggaatcg atcctcccaa agagaaatag cgacaagctc attgcacgca agaaagactg     3480

ggacccgaag aagtacggag gattcgattc gccgactgtc gcatactccg tcctcgtggt     3540

ggccaaggtg gagaagggaa agagcaaaaa gctcaaatcc gtcaaagagc tgctggggat     3600

taccatcatg gaacgatcct cgttcgagaa gaacccgatt gatttcctcg aggcgaaggg     3660

ttacaaggag gtgaagaagg atctgatcat caaactcccc aagtactcac tgttcgaact     3720

ggaaaatggt cggaagcgca tgctggcttc ggccggagaa ctccaaaaag gaaatgagct     3780

ggccttgcct agcaagtacg tcaacttcct ctatcttgct tcgcactacg aaaaactcaa     3840

agggtcaccg gaagataacg aacagaagca gcttttcgtg gagcagcaca agcattatct     3900

ggatgaaatc atcgaacaaa tctccgagtt ttcaaagcgc gtgatcctcg ccgacgccaa     3960

cctcgacaaa gtcctgtcgg cctacaataa gcatagagat aagccgatca gagaacaggc     4020

cgagaacatt atccacttgt tcaccctgac taacctggga gccccagccg ccttcaagta     4080

cttcgatact actatcgatc gcaaaagata cacgtccacc aaggaagttc tggacgcgac     4140

cctgatccac caaagcatca ctggactcta cgaaactagg atcgatctgt cgcagctggg     4200

tggcgattga tagtctagcc atcacattta aaagcatctc agcctaccat gagaataaga     4260

gaaagaaaat gaagatcaat agcttattca tctctttttc tttttcgttg gtgtaaagcc     4320

aacaccctgt ctaaaaaaca taaatttctt taatcatttt gcctcttttc tctgtgcttc     4380

aattaataaa aaatggaaag aacctcgag                                       4409


<210>  250
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF with splice junctions removed; 12.75% U 
       content

<400>  250
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccaccgg      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca aaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcccga agaagaagag aaaggtctag     4140


<210>  251
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 250, Kozak sequence, and 3 UTR of ALB

<400>  251
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc caccggctgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcaaaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  252
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF with minimal uridine codons frequently used 
       in humans in general; 12.75% U content

<400>  252
atggacaaga agtacagcat cggcctggac atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgacaga      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccaccagac tgaagagaac cgccagaaga agatacacca gaagaaagaa cagaatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agagacaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aagctggtgg acagcaccga caaggccgac ctgagactga tctacctggc cctggcccac      480

atgatcaagt tcagaggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg ccagactgag caagagcaga      660

agactggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgaga     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga acagagagga cctgctgaga     1200

aagcagagaa ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctga gaagacagga ggacttctac cccttcctga aggacaacag agagaagatc     1320

gagaagatcc tgaccttcag aatcccctac tacgtgggcc ccctggccag aggcaacagc     1380

agattcgcct ggatgaccag aaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgaga gaatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgagaaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaacag aaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggacagatt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggacagagag atgatcgagg agagactgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagagaa gaagatacac cggctggggc     1980

agactgagca gaaagctgat caacggcatc agagacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggcagac acaagcccga gaacatcgtg     2280

atcgagatgg ccagagagaa ccagaccacc cagaagggcc agaagaacag cagagagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggcaga     2460

gacatgtacg tggaccagga gctggacatc aacagactga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gaccagaagc     2580

gacaagaaca gaggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactgga gacagctgct gaacgccaag ctgatcaccc agagaaagtt cgacaacctg     2700

accaaggccg agagaggcgg cctgagcgag ctggacaagg ccggcttcat caagagacag     2760

ctggtggaga ccagacagat caccaagcac gtggcccaga tcctggacag cagaatgaac     2820

accaagtacg acgagaacga caagctgatc agagaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttcag aaaggacttc cagttctaca aggtgagaga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgagaaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat cagaaagaga     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg cagagacttc     3240

gccaccgtga gaaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaaga gaaacagcga caagctgatc     3360

gccagaaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag agaagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggcaga aagagaatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagagagtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca cagagacaag     3900

cccatcagag agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgacagaa agagatacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gaccagaatc     4080

gacctgagcc agctgggcgg cgacggcggc ggcagcccca agaagaagag aaaggtgtga     4140


<210>  253
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 252, Kozak sequence, and 3 UTR of ALB

<400>  253
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcggc ctggacatcg gcaccaacag      120

cgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaagt tcaaggtgct      180

gggcaacacc gacagacaca gcatcaagaa gaacctgatc ggcgccctgc tgttcgacag      240

cggcgagacc gccgaggcca ccagactgaa gagaaccgcc agaagaagat acaccagaag      300

aaagaacaga atctgctacc tgcaggagat cttcagcaac gagatggcca aggtggacga      360

cagcttcttc cacagactgg aggagagctt cctggtggag gaggacaaga agcacgagag      420

acaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat      480

ctaccacctg agaaagaagc tggtggacag caccgacaag gccgacctga gactgatcta      540

cctggccctg gcccacatga tcaagttcag aggccacttc ctgatcgagg gcgacctgaa      600

ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct      660

gttcgaggag aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgagcgccag      720

actgagcaag agcagaagac tggagaacct gatcgcccag ctgcccggcg agaagaagaa      780

cggcctgttc ggcaacctga tcgccctgag cctgggcctg acccccaact tcaagagcaa      840

cttcgacctg gccgaggacg ccaagctgca gctgagcaag gacacctacg acgacgacct      900

ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgttcctgg ccgccaagaa      960

cctgagcgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc     1020

ccccctgagc gccagcatga tcaagagata cgacgagcac caccaggacc tgaccctgct     1080

gaaggccctg gtgagacagc agctgcccga gaagtacaag gagatcttct tcgaccagag     1140

caagaacggc tacgccggct acatcgacgg cggcgccagc caggaggagt tctacaagtt     1200

catcaagccc atcctggaga agatggacgg caccgaggag ctgctggtga agctgaacag     1260

agaggacctg ctgagaaagc agagaacctt cgacaacggc agcatccccc accagatcca     1320

cctgggcgag ctgcacgcca tcctgagaag acaggaggac ttctacccct tcctgaagga     1380

caacagagag aagatcgaga agatcctgac cttcagaatc ccctactacg tgggccccct     1440

ggccagaggc aacagcagat tcgcctggat gaccagaaag agcgaggaga ccatcacccc     1500

ctggaacttc gaggaggtgg tggacaaggg cgccagcgcc cagagcttca tcgagagaat     1560

gaccaacttc gacaagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta     1620

cgagtacttc accgtgtaca acgagctgac caaggtgaag tacgtgaccg agggcatgag     1680

aaagcccgcc ttcctgagcg gcgagcagaa gaaggccatc gtggacctgc tgttcaagac     1740

caacagaaag gtgaccgtga agcagctgaa ggaggactac ttcaagaaga tcgagtgctt     1800

cgacagcgtg gagatcagcg gcgtggagga cagattcaac gccagcctgg gcacctacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaggaga acgaggacat     1920

cctggaggac atcgtgctga ccctgaccct gttcgaggac agagagatga tcgaggagag     1980

actgaagacc tacgcccacc tgttcgacga caaggtgatg aagcagctga agagaagaag     2040

atacaccggc tggggcagac tgagcagaaa gctgatcaac ggcatcagag acaagcagag     2100

cggcaagacc atcctggact tcctgaagag cgacggcttc gccaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgaccttcaa ggaggacatc cagaaggccc aggtgagcgg     2220

ccagggcgac agcctgcacg agcacatcgc caacctggcc ggcagccccg ccatcaagaa     2280

gggcatcctg cagaccgtga aggtggtgga cgagctggtg aaggtgatgg gcagacacaa     2340

gcccgagaac atcgtgatcg agatggccag agagaaccag accacccaga agggccagaa     2400

gaacagcaga gagagaatga agagaatcga ggagggcatc aaggagctgg gcagccagat     2460

cctgaaggag caccccgtgg agaacaccca gctgcagaac gagaagctgt acctgtacta     2520

cctgcagaac ggcagagaca tgtacgtgga ccaggagctg gacatcaaca gactgagcga     2580

ctacgacgtg gaccacatcg tgccccagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtgctgacc agaagcgaca agaacagagg caagagcgac aacgtgccca gcgaggaggt     2700

ggtgaagaag atgaagaact actggagaca gctgctgaac gccaagctga tcacccagag     2760

aaagttcgac aacctgacca aggccgagag aggcggcctg agcgagctgg acaaggccgg     2820

cttcatcaag agacagctgg tggagaccag acagatcacc aagcacgtgg cccagatcct     2880

ggacagcaga atgaacacca agtacgacga gaacgacaag ctgatcagag aggtgaaggt     2940

gatcaccctg aagagcaagc tggtgagcga cttcagaaag gacttccagt tctacaaggt     3000

gagagagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tggtgggcac     3060

cgccctgatc aagaagtacc ccaagctgga gagcgagttc gtgtacggcg actacaaggt     3120

gtacgacgtg agaaagatga tcgccaagag cgagcaggag atcggcaagg ccaccgccaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagacc gagatcaccc tggccaacgg     3240

cgagatcaga aagagacccc tgatcgagac caacggcgag accggcgaga tcgtgtggga     3300

caagggcaga gacttcgcca ccgtgagaaa ggtgctgagc atgccccagg tgaacatcgt     3360

gaagaagacc gaggtgcaga ccggcggctt cagcaaggag agcatcctgc ccaagagaaa     3420

cagcgacaag ctgatcgcca gaaagaagga ctgggacccc aagaagtacg gcggcttcga     3480

cagccccacc gtggcctaca gcgtgctggt ggtggccaag gtggagaagg gcaagagcaa     3540

gaagctgaag agcgtgaagg agctgctggg catcaccatc atggagagaa gcagcttcga     3600

gaagaacccc atcgacttcc tggaggccaa gggctacaag gaggtgaaga aggacctgat     3660

catcaagctg cccaagtaca gcctgttcga gctggagaac ggcagaaaga gaatgctggc     3720

cagcgccggc gagctgcaga agggcaacga gctggccctg cccagcaagt acgtgaactt     3780

cctgtacctg gccagccact acgagaagct gaagggcagc cccgaggaca acgagcagaa     3840

gcagctgttc gtggagcagc acaagcacta cctggacgag atcatcgagc agatcagcga     3900

gttcagcaag agagtgatcc tggccgacgc caacctggac aaggtgctga gcgcctacaa     3960

caagcacaga gacaagccca tcagagagca ggccgagaac atcatccacc tgttcaccct     4020

gaccaacctg ggcgcccccg ccgccttcaa gtacttcgac accaccatcg acagaaagag     4080

atacaccagc accaaggagg tgctggacgc caccctgatc caccagagca tcaccggcct     4140

gtacgagacc agaatcgacc tgagccagct gggcggcgac ggcggcggca gccccaagaa     4200

gaagagaaag gtgtgactag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  254
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF with minimal uridine codons infrequently used
       in humans in general; 12.75% U content

<400>  254
atggacaaaa aatacagcat agggctagac atagggacga acagcgtagg gtgggcggta       60

ataacggacg aatacaaagt accgagcaaa aaattcaaag tactagggaa cacggaccga      120

cacagcataa aaaaaaacct aataggggcg ctactattcg acagcgggga aacggcggaa      180

gcgacgcgac taaaacgaac ggcgcgacga cgatacacgc gacgaaaaaa ccgaatatgc      240

tacctacaag aaatattcag caacgaaatg gcgaaagtag acgacagctt cttccaccga      300

ctagaagaaa gcttcctagt agaagaagac aaaaaacacg aacgacaccc gatattcggg      360

aacatagtag acgaagtagc gtaccacgaa aaatacccga cgatatacca cctacgaaaa      420

aaactagtag acagcacgga caaagcggac ctacgactaa tatacctagc gctagcgcac      480

atgataaaat tccgagggca cttcctaata gaaggggacc taaacccgga caacagcgac      540

gtagacaaac tattcataca actagtacaa acgtacaacc aactattcga agaaaacccg      600

ataaacgcga gcggggtaga cgcgaaagcg atactaagcg cgcgactaag caaaagccga      660

cgactagaaa acctaatagc gcaactaccg ggggaaaaaa aaaacgggct attcgggaac      720

ctaatagcgc taagcctagg gctaacgccg aacttcaaaa gcaacttcga cctagcggaa      780

gacgcgaaac tacaactaag caaagacacg tacgacgacg acctagacaa cctactagcg      840

caaatagggg accaatacgc ggacctattc ctagcggcga aaaacctaag cgacgcgata      900

ctactaagcg acatactacg agtaaacacg gaaataacga aagcgccgct aagcgcgagc      960

atgataaaac gatacgacga acaccaccaa gacctaacgc tactaaaagc gctagtacga     1020

caacaactac cggaaaaata caaagaaata ttcttcgacc aaagcaaaaa cgggtacgcg     1080

gggtacatag acgggggggc gagccaagaa gaattctaca aattcataaa accgatacta     1140

gaaaaaatgg acgggacgga agaactacta gtaaaactaa accgagaaga cctactacga     1200

aaacaacgaa cgttcgacaa cgggagcata ccgcaccaaa tacacctagg ggaactacac     1260

gcgatactac gacgacaaga agacttctac ccgttcctaa aagacaaccg agaaaaaata     1320

gaaaaaatac taacgttccg aataccgtac tacgtagggc cgctagcgcg agggaacagc     1380

cgattcgcgt ggatgacgcg aaaaagcgaa gaaacgataa cgccgtggaa cttcgaagaa     1440

gtagtagaca aaggggcgag cgcgcaaagc ttcatagaac gaatgacgaa cttcgacaaa     1500

aacctaccga acgaaaaagt actaccgaaa cacagcctac tatacgaata cttcacggta     1560

tacaacgaac taacgaaagt aaaatacgta acggaaggga tgcgaaaacc ggcgttccta     1620

agcggggaac aaaaaaaagc gatagtagac ctactattca aaacgaaccg aaaagtaacg     1680

gtaaaacaac taaaagaaga ctacttcaaa aaaatagaat gcttcgacag cgtagaaata     1740

agcggggtag aagaccgatt caacgcgagc ctagggacgt accacgacct actaaaaata     1800

ataaaagaca aagacttcct agacaacgaa gaaaacgaag acatactaga agacatagta     1860

ctaacgctaa cgctattcga agaccgagaa atgatagaag aacgactaaa aacgtacgcg     1920

cacctattcg acgacaaagt aatgaaacaa ctaaaacgac gacgatacac ggggtggggg     1980

cgactaagcc gaaaactaat aaacgggata cgagacaaac aaagcgggaa aacgatacta     2040

gacttcctaa aaagcgacgg gttcgcgaac cgaaacttca tgcaactaat acacgacgac     2100

agcctaacgt tcaaagaaga catacaaaaa gcgcaagtaa gcgggcaagg ggacagccta     2160

cacgaacaca tagcgaacct agcggggagc ccggcgataa aaaaagggat actacaaacg     2220

gtaaaagtag tagacgaact agtaaaagta atggggcgac acaaaccgga aaacatagta     2280

atagaaatgg cgcgagaaaa ccaaacgacg caaaaagggc aaaaaaacag ccgagaacga     2340

atgaaacgaa tagaagaagg gataaaagaa ctagggagcc aaatactaaa agaacacccg     2400

gtagaaaaca cgcaactaca aaacgaaaaa ctatacctat actacctaca aaacgggcga     2460

gacatgtacg tagaccaaga actagacata aaccgactaa gcgactacga cgtagaccac     2520

atagtaccgc aaagcttcct aaaagacgac agcatagaca acaaagtact aacgcgaagc     2580

gacaaaaacc gagggaaaag cgacaacgta ccgagcgaag aagtagtaaa aaaaatgaaa     2640

aactactggc gacaactact aaacgcgaaa ctaataacgc aacgaaaatt cgacaaccta     2700

acgaaagcgg aacgaggggg gctaagcgaa ctagacaaag cggggttcat aaaacgacaa     2760

ctagtagaaa cgcgacaaat aacgaaacac gtagcgcaaa tactagacag ccgaatgaac     2820

acgaaatacg acgaaaacga caaactaata cgagaagtaa aagtaataac gctaaaaagc     2880

aaactagtaa gcgacttccg aaaagacttc caattctaca aagtacgaga aataaacaac     2940

taccaccacg cgcacgacgc gtacctaaac gcggtagtag ggacggcgct aataaaaaaa     3000

tacccgaaac tagaaagcga attcgtatac ggggactaca aagtatacga cgtacgaaaa     3060

atgatagcga aaagcgaaca agaaataggg aaagcgacgg cgaaatactt cttctacagc     3120

aacataatga acttcttcaa aacggaaata acgctagcga acggggaaat acgaaaacga     3180

ccgctaatag aaacgaacgg ggaaacgggg gaaatagtat gggacaaagg gcgagacttc     3240

gcgacggtac gaaaagtact aagcatgccg caagtaaaca tagtaaaaaa aacggaagta     3300

caaacggggg ggttcagcaa agaaagcata ctaccgaaac gaaacagcga caaactaata     3360

gcgcgaaaaa aagactggga cccgaaaaaa tacggggggt tcgacagccc gacggtagcg     3420

tacagcgtac tagtagtagc gaaagtagaa aaagggaaaa gcaaaaaact aaaaagcgta     3480

aaagaactac tagggataac gataatggaa cgaagcagct tcgaaaaaaa cccgatagac     3540

ttcctagaag cgaaagggta caaagaagta aaaaaagacc taataataaa actaccgaaa     3600

tacagcctat tcgaactaga aaacgggcga aaacgaatgc tagcgagcgc gggggaacta     3660

caaaaaggga acgaactagc gctaccgagc aaatacgtaa acttcctata cctagcgagc     3720

cactacgaaa aactaaaagg gagcccggaa gacaacgaac aaaaacaact attcgtagaa     3780

caacacaaac actacctaga cgaaataata gaacaaataa gcgaattcag caaacgagta     3840

atactagcgg acgcgaacct agacaaagta ctaagcgcgt acaacaaaca ccgagacaaa     3900

ccgatacgag aacaagcgga aaacataata cacctattca cgctaacgaa cctaggggcg     3960

ccggcggcgt tcaaatactt cgacacgacg atagaccgaa aacgatacac gagcacgaaa     4020

gaagtactag acgcgacgct aatacaccaa agcataacgg ggctatacga aacgcgaata     4080

gacctaagcc aactaggggg ggacgggggg gggagcccga aaaaaaaacg aaaagtatga     4140


<210>  255
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 254, Kozak sequence, and 3 UTR of ALB

<400>  255
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaaaaaata cagcataggg ctagacatag ggacgaacag      120

cgtagggtgg gcggtaataa cggacgaata caaagtaccg agcaaaaaat tcaaagtact      180

agggaacacg gaccgacaca gcataaaaaa aaacctaata ggggcgctac tattcgacag      240

cggggaaacg gcggaagcga cgcgactaaa acgaacggcg cgacgacgat acacgcgacg      300

aaaaaaccga atatgctacc tacaagaaat attcagcaac gaaatggcga aagtagacga      360

cagcttcttc caccgactag aagaaagctt cctagtagaa gaagacaaaa aacacgaacg      420

acacccgata ttcgggaaca tagtagacga agtagcgtac cacgaaaaat acccgacgat      480

ataccaccta cgaaaaaaac tagtagacag cacggacaaa gcggacctac gactaatata      540

cctagcgcta gcgcacatga taaaattccg agggcacttc ctaatagaag gggacctaaa      600

cccggacaac agcgacgtag acaaactatt catacaacta gtacaaacgt acaaccaact      660

attcgaagaa aacccgataa acgcgagcgg ggtagacgcg aaagcgatac taagcgcgcg      720

actaagcaaa agccgacgac tagaaaacct aatagcgcaa ctaccggggg aaaaaaaaaa      780

cgggctattc gggaacctaa tagcgctaag cctagggcta acgccgaact tcaaaagcaa      840

cttcgaccta gcggaagacg cgaaactaca actaagcaaa gacacgtacg acgacgacct      900

agacaaccta ctagcgcaaa taggggacca atacgcggac ctattcctag cggcgaaaaa      960

cctaagcgac gcgatactac taagcgacat actacgagta aacacggaaa taacgaaagc     1020

gccgctaagc gcgagcatga taaaacgata cgacgaacac caccaagacc taacgctact     1080

aaaagcgcta gtacgacaac aactaccgga aaaatacaaa gaaatattct tcgaccaaag     1140

caaaaacggg tacgcggggt acatagacgg gggggcgagc caagaagaat tctacaaatt     1200

cataaaaccg atactagaaa aaatggacgg gacggaagaa ctactagtaa aactaaaccg     1260

agaagaccta ctacgaaaac aacgaacgtt cgacaacggg agcataccgc accaaataca     1320

cctaggggaa ctacacgcga tactacgacg acaagaagac ttctacccgt tcctaaaaga     1380

caaccgagaa aaaatagaaa aaatactaac gttccgaata ccgtactacg tagggccgct     1440

agcgcgaggg aacagccgat tcgcgtggat gacgcgaaaa agcgaagaaa cgataacgcc     1500

gtggaacttc gaagaagtag tagacaaagg ggcgagcgcg caaagcttca tagaacgaat     1560

gacgaacttc gacaaaaacc taccgaacga aaaagtacta ccgaaacaca gcctactata     1620

cgaatacttc acggtataca acgaactaac gaaagtaaaa tacgtaacgg aagggatgcg     1680

aaaaccggcg ttcctaagcg gggaacaaaa aaaagcgata gtagacctac tattcaaaac     1740

gaaccgaaaa gtaacggtaa aacaactaaa agaagactac ttcaaaaaaa tagaatgctt     1800

cgacagcgta gaaataagcg gggtagaaga ccgattcaac gcgagcctag ggacgtacca     1860

cgacctacta aaaataataa aagacaaaga cttcctagac aacgaagaaa acgaagacat     1920

actagaagac atagtactaa cgctaacgct attcgaagac cgagaaatga tagaagaacg     1980

actaaaaacg tacgcgcacc tattcgacga caaagtaatg aaacaactaa aacgacgacg     2040

atacacgggg tgggggcgac taagccgaaa actaataaac gggatacgag acaaacaaag     2100

cgggaaaacg atactagact tcctaaaaag cgacgggttc gcgaaccgaa acttcatgca     2160

actaatacac gacgacagcc taacgttcaa agaagacata caaaaagcgc aagtaagcgg     2220

gcaaggggac agcctacacg aacacatagc gaacctagcg gggagcccgg cgataaaaaa     2280

agggatacta caaacggtaa aagtagtaga cgaactagta aaagtaatgg ggcgacacaa     2340

accggaaaac atagtaatag aaatggcgcg agaaaaccaa acgacgcaaa aagggcaaaa     2400

aaacagccga gaacgaatga aacgaataga agaagggata aaagaactag ggagccaaat     2460

actaaaagaa cacccggtag aaaacacgca actacaaaac gaaaaactat acctatacta     2520

cctacaaaac gggcgagaca tgtacgtaga ccaagaacta gacataaacc gactaagcga     2580

ctacgacgta gaccacatag taccgcaaag cttcctaaaa gacgacagca tagacaacaa     2640

agtactaacg cgaagcgaca aaaaccgagg gaaaagcgac aacgtaccga gcgaagaagt     2700

agtaaaaaaa atgaaaaact actggcgaca actactaaac gcgaaactaa taacgcaacg     2760

aaaattcgac aacctaacga aagcggaacg aggggggcta agcgaactag acaaagcggg     2820

gttcataaaa cgacaactag tagaaacgcg acaaataacg aaacacgtag cgcaaatact     2880

agacagccga atgaacacga aatacgacga aaacgacaaa ctaatacgag aagtaaaagt     2940

aataacgcta aaaagcaaac tagtaagcga cttccgaaaa gacttccaat tctacaaagt     3000

acgagaaata aacaactacc accacgcgca cgacgcgtac ctaaacgcgg tagtagggac     3060

ggcgctaata aaaaaatacc cgaaactaga aagcgaattc gtatacgggg actacaaagt     3120

atacgacgta cgaaaaatga tagcgaaaag cgaacaagaa atagggaaag cgacggcgaa     3180

atacttcttc tacagcaaca taatgaactt cttcaaaacg gaaataacgc tagcgaacgg     3240

ggaaatacga aaacgaccgc taatagaaac gaacggggaa acgggggaaa tagtatggga     3300

caaagggcga gacttcgcga cggtacgaaa agtactaagc atgccgcaag taaacatagt     3360

aaaaaaaacg gaagtacaaa cgggggggtt cagcaaagaa agcatactac cgaaacgaaa     3420

cagcgacaaa ctaatagcgc gaaaaaaaga ctgggacccg aaaaaatacg gggggttcga     3480

cagcccgacg gtagcgtaca gcgtactagt agtagcgaaa gtagaaaaag ggaaaagcaa     3540

aaaactaaaa agcgtaaaag aactactagg gataacgata atggaacgaa gcagcttcga     3600

aaaaaacccg atagacttcc tagaagcgaa agggtacaaa gaagtaaaaa aagacctaat     3660

aataaaacta ccgaaataca gcctattcga actagaaaac gggcgaaaac gaatgctagc     3720

gagcgcgggg gaactacaaa aagggaacga actagcgcta ccgagcaaat acgtaaactt     3780

cctataccta gcgagccact acgaaaaact aaaagggagc ccggaagaca acgaacaaaa     3840

acaactattc gtagaacaac acaaacacta cctagacgaa ataatagaac aaataagcga     3900

attcagcaaa cgagtaatac tagcggacgc gaacctagac aaagtactaa gcgcgtacaa     3960

caaacaccga gacaaaccga tacgagaaca agcggaaaac ataatacacc tattcacgct     4020

aacgaaccta ggggcgccgg cggcgttcaa atacttcgac acgacgatag accgaaaacg     4080

atacacgagc acgaaagaag tactagacgc gacgctaata caccaaagca taacggggct     4140

atacgaaacg cgaatagacc taagccaact agggggggac ggggggggga gcccgaaaaa     4200

aaaacgaaaa gtatgactag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  256
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with AGG as first three nucleotides 
       for use with CleanCapTM, 5 UTR of HSD, ORF corresponding to SEQ 
       ID NO: 204, Kozak sequence, and 3 UTR of ALB

<400>  256
aggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  257
<211>  4481
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR from CMV, ORF corresponding
       to SEQ ID NO: 204, Kozak sequence, and 3 UTR of ALB

<400>  257
gggcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac       60

cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt      120

gactcaccgt ccttgacacg gccaccatgg acaagaagta cagcatcgga ctggacatcg      180

gaacaaacag cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt      240

tcaaggtcct gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc      300

tgttcgacag cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat      360

acacaagaag aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa      420

aggtcgacga cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga      480

agcacgaaag acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt      540

acccgacaat ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga      600

gactgatcta cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag      660

gagacctgaa cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat      720

acaaccagct gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc      780

tgagcgcaag actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag      840

aaaagaagaa cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact      900

tcaagagcaa cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg      960

acgacgacct ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg     1020

cagcaaagaa cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa     1080

tcacaaaggc accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc     1140

tgacactgct gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct     1200

tcgaccagag caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat     1260

tctacaagtt catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca     1320

agctgaacag agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc     1380

accagatcca cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt     1440

tcctgaagga caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg     1500

tcggaccgct ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa     1560

caatcacacc gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca     1620

tcgaaagaat gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca     1680

gcctgctgta cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag     1740

aaggaatgag aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc     1800

tgttcaagac aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga     1860

tcgaatgctt cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg     1920

gaacatacca cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa     1980

acgaagacat cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga     2040

tcgaagaaag actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga     2100

agagaagaag atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag     2160

acaagcagag cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa     2220

acttcatgca gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac     2280

aggtcagcgg acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg     2340

caatcaagaa gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg     2400

gaagacacaa gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga     2460

agggacagaa gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg     2520

gaagccagat cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt     2580

acctgtacta cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca     2640

gactgagcga ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca     2700

tcgacaacaa ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga     2760

gcgaagaagt cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga     2820

tcacacagag aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg     2880

acaaggcagg attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg     2940

cacagatcct ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag     3000

aagtcaaggt catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt     3060

tctacaaggt cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag     3120

tcgtcggaac agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag     3180

actacaaggt ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg     3240

caacagcaaa gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac     3300

tggcaaacgg agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa     3360

tcgtctggga caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg     3420

tcaacatcgt caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc     3480

cgaagagaaa cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg     3540

gaggattcga cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg     3600

gaaagagcaa gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa     3660

gcagcttcga aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga     3720

aggacctgat catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga     3780

gaatgctggc aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt     3840

acgtcaactt cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca     3900

acgaacagaa gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac     3960

agatcagcga attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga     4020

gcgcatacaa caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc     4080

tgttcacact gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg     4140

acagaaagag atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca     4200

tcacaggact gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa     4260

gcccgaagaa gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc     4320

atgagaataa gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt     4380

tggtgtaaag ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt     4440

tctctgtgct tcaattaata aaaaatggaa agaacctcga g                         4481


<210>  258
<211>  4348
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR from HBB, ORF corresponding
       to SEQ ID NO: 204, Kozak sequence, and 3 UTR of HBB

<400>  258
gggacatttg cttctgacac aactgtgttc actagcaacc tcaaacagac accggatctg       60

ccaccatgga caagaagtac agcatcggac tggacatcgg aacaaacagc gtcggatggg      120

cagtcatcac agacgaatac aaggtcccga gcaagaagtt caaggtcctg ggaaacacag      180

acagacacag catcaagaag aacctgatcg gagcactgct gttcgacagc ggagaaacag      240

cagaagcaac aagactgaag agaacagcaa gaagaagata cacaagaaga aagaacagaa      300

tctgctacct gcaggaaatc ttcagcaacg aaatggcaaa ggtcgacgac agcttcttcc      360

acagactgga agaaagcttc ctggtcgaag aagacaagaa gcacgaaaga cacccgatct      420

tcggaaacat cgtcgacgaa gtcgcatacc acgaaaagta cccgacaatc taccacctga      480

gaaagaagct ggtcgacagc acagacaagg cagacctgag actgatctac ctggcactgg      540

cacacatgat caagttcaga ggacacttcc tgatcgaagg agacctgaac ccggacaaca      600

gcgacgtcga caagctgttc atccagctgg tccagacata caaccagctg ttcgaagaaa      660

acccgatcaa cgcaagcgga gtcgacgcaa aggcaatcct gagcgcaaga ctgagcaaga      720

gcagaagact ggaaaacctg atcgcacagc tgccgggaga aaagaagaac ggactgttcg      780

gaaacctgat cgcactgagc ctgggactga caccgaactt caagagcaac ttcgacctgg      840

cagaagacgc aaagctgcag ctgagcaagg acacatacga cgacgacctg gacaacctgc      900

tggcacagat cggagaccag tacgcagacc tgttcctggc agcaaagaac ctgagcgacg      960

caatcctgct gagcgacatc ctgagagtca acacagaaat cacaaaggca ccgctgagcg     1020

caagcatgat caagagatac gacgaacacc accaggacct gacactgctg aaggcactgg     1080

tcagacagca gctgccggaa aagtacaagg aaatcttctt cgaccagagc aagaacggat     1140

acgcaggata catcgacgga ggagcaagcc aggaagaatt ctacaagttc atcaagccga     1200

tcctggaaaa gatggacgga acagaagaac tgctggtcaa gctgaacaga gaagacctgc     1260

tgagaaagca gagaacattc gacaacggaa gcatcccgca ccagatccac ctgggagaac     1320

tgcacgcaat cctgagaaga caggaagact tctacccgtt cctgaaggac aacagagaaa     1380

agatcgaaaa gatcctgaca ttcagaatcc cgtactacgt cggaccgctg gcaagaggaa     1440

acagcagatt cgcatggatg acaagaaaga gcgaagaaac aatcacaccg tggaacttcg     1500

aagaagtcgt cgacaaggga gcaagcgcac agagcttcat cgaaagaatg acaaacttcg     1560

acaagaacct gccgaacgaa aaggtcctgc cgaagcacag cctgctgtac gaatacttca     1620

cagtctacaa cgaactgaca aaggtcaagt acgtcacaga aggaatgaga aagccggcat     1680

tcctgagcgg agaacagaag aaggcaatcg tcgacctgct gttcaagaca aacagaaagg     1740

tcacagtcaa gcagctgaag gaagactact tcaagaagat cgaatgcttc gacagcgtcg     1800

aaatcagcgg agtcgaagac agattcaacg caagcctggg aacataccac gacctgctga     1860

agatcatcaa ggacaaggac ttcctggaca acgaagaaaa cgaagacatc ctggaagaca     1920

tcgtcctgac actgacactg ttcgaagaca gagaaatgat cgaagaaaga ctgaagacat     1980

acgcacacct gttcgacgac aaggtcatga agcagctgaa gagaagaaga tacacaggat     2040

ggggaagact gagcagaaag ctgatcaacg gaatcagaga caagcagagc ggaaagacaa     2100

tcctggactt cctgaagagc gacggattcg caaacagaaa cttcatgcag ctgatccacg     2160

acgacagcct gacattcaag gaagacatcc agaaggcaca ggtcagcgga cagggagaca     2220

gcctgcacga acacatcgca aacctggcag gaagcccggc aatcaagaag ggaatcctgc     2280

agacagtcaa ggtcgtcgac gaactggtca aggtcatggg aagacacaag ccggaaaaca     2340

tcgtcatcga aatggcaaga gaaaaccaga caacacagaa gggacagaag aacagcagag     2400

aaagaatgaa gagaatcgaa gaaggaatca aggaactggg aagccagatc ctgaaggaac     2460

acccggtcga aaacacacag ctgcagaacg aaaagctgta cctgtactac ctgcagaacg     2520

gaagagacat gtacgtcgac caggaactgg acatcaacag actgagcgac tacgacgtcg     2580

accacatcgt cccgcagagc ttcctgaagg acgacagcat cgacaacaag gtcctgacaa     2640

gaagcgacaa gaacagagga aagagcgaca acgtcccgag cgaagaagtc gtcaagaaga     2700

tgaagaacta ctggagacag ctgctgaacg caaagctgat cacacagaga aagttcgaca     2760

acctgacaaa ggcagagaga ggaggactga gcgaactgga caaggcagga ttcatcaaga     2820

gacagctggt cgaaacaaga cagatcacaa agcacgtcgc acagatcctg gacagcagaa     2880

tgaacacaaa gtacgacgaa aacgacaagc tgatcagaga agtcaaggtc atcacactga     2940

agagcaagct ggtcagcgac ttcagaaagg acttccagtt ctacaaggtc agagaaatca     3000

acaactacca ccacgcacac gacgcatacc tgaacgcagt cgtcggaaca gcactgatca     3060

agaagtaccc gaagctggaa agcgaattcg tctacggaga ctacaaggtc tacgacgtca     3120

gaaagatgat cgcaaagagc gaacaggaaa tcggaaaggc aacagcaaag tacttcttct     3180

acagcaacat catgaacttc ttcaagacag aaatcacact ggcaaacgga gaaatcagaa     3240

agagaccgct gatcgaaaca aacggagaaa caggagaaat cgtctgggac aagggaagag     3300

acttcgcaac agtcagaaag gtcctgagca tgccgcaggt caacatcgtc aagaagacag     3360

aagtccagac aggaggattc agcaaggaaa gcatcctgcc gaagagaaac agcgacaagc     3420

tgatcgcaag aaagaaggac tgggacccga agaagtacgg aggattcgac agcccgacag     3480

tcgcatacag cgtcctggtc gtcgcaaagg tcgaaaaggg aaagagcaag aagctgaaga     3540

gcgtcaagga actgctggga atcacaatca tggaaagaag cagcttcgaa aagaacccga     3600

tcgacttcct ggaagcaaag ggatacaagg aagtcaagaa ggacctgatc atcaagctgc     3660

cgaagtacag cctgttcgaa ctggaaaacg gaagaaagag aatgctggca agcgcaggag     3720

aactgcagaa gggaaacgaa ctggcactgc cgagcaagta cgtcaacttc ctgtacctgg     3780

caagccacta cgaaaagctg aagggaagcc cggaagacaa cgaacagaag cagctgttcg     3840

tcgaacagca caagcactac ctggacgaaa tcatcgaaca gatcagcgaa ttcagcaaga     3900

gagtcatcct ggcagacgca aacctggaca aggtcctgag cgcatacaac aagcacagag     3960

acaagccgat cagagaacag gcagaaaaca tcatccacct gttcacactg acaaacctgg     4020

gagcaccggc agcattcaag tacttcgaca caacaatcga cagaaagaga tacacaagca     4080

caaaggaagt cctggacgca acactgatcc accagagcat cacaggactg tacgaaacaa     4140

gaatcgacct gagccagctg ggaggagacg gaggaggaag cccgaagaag aagagaaagg     4200

tctagctagc gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa     4260

gtccaactac taaactgggg gatattatga agggccttga gcatctggat tctgcctaat     4320

aaaaaacatt tattttcatt gcctcgag                                        4348


<210>  259
<211>  4325
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR from XBG, ORF corresponding
       to SEQ ID NO: 204, Kozak sequence, and 3 UTR of XBG

<400>  259
gggaagctca gaataaacgc tcaactttgg ccggatctgc caccatggac aagaagtaca       60

gcatcggact ggacatcgga acaaacagcg tcggatgggc agtcatcaca gacgaataca      120

aggtcccgag caagaagttc aaggtcctgg gaaacacaga cagacacagc atcaagaaga      180

acctgatcgg agcactgctg ttcgacagcg gagaaacagc agaagcaaca agactgaaga      240

gaacagcaag aagaagatac acaagaagaa agaacagaat ctgctacctg caggaaatct      300

tcagcaacga aatggcaaag gtcgacgaca gcttcttcca cagactggaa gaaagcttcc      360

tggtcgaaga agacaagaag cacgaaagac acccgatctt cggaaacatc gtcgacgaag      420

tcgcatacca cgaaaagtac ccgacaatct accacctgag aaagaagctg gtcgacagca      480

cagacaaggc agacctgaga ctgatctacc tggcactggc acacatgatc aagttcagag      540

gacacttcct gatcgaagga gacctgaacc cggacaacag cgacgtcgac aagctgttca      600

tccagctggt ccagacatac aaccagctgt tcgaagaaaa cccgatcaac gcaagcggag      660

tcgacgcaaa ggcaatcctg agcgcaagac tgagcaagag cagaagactg gaaaacctga      720

tcgcacagct gccgggagaa aagaagaacg gactgttcgg aaacctgatc gcactgagcc      780

tgggactgac accgaacttc aagagcaact tcgacctggc agaagacgca aagctgcagc      840

tgagcaagga cacatacgac gacgacctgg acaacctgct ggcacagatc ggagaccagt      900

acgcagacct gttcctggca gcaaagaacc tgagcgacgc aatcctgctg agcgacatcc      960

tgagagtcaa cacagaaatc acaaaggcac cgctgagcgc aagcatgatc aagagatacg     1020

acgaacacca ccaggacctg acactgctga aggcactggt cagacagcag ctgccggaaa     1080

agtacaagga aatcttcttc gaccagagca agaacggata cgcaggatac atcgacggag     1140

gagcaagcca ggaagaattc tacaagttca tcaagccgat cctggaaaag atggacggaa     1200

cagaagaact gctggtcaag ctgaacagag aagacctgct gagaaagcag agaacattcg     1260

acaacggaag catcccgcac cagatccacc tgggagaact gcacgcaatc ctgagaagac     1320

aggaagactt ctacccgttc ctgaaggaca acagagaaaa gatcgaaaag atcctgacat     1380

tcagaatccc gtactacgtc ggaccgctgg caagaggaaa cagcagattc gcatggatga     1440

caagaaagag cgaagaaaca atcacaccgt ggaacttcga agaagtcgtc gacaagggag     1500

caagcgcaca gagcttcatc gaaagaatga caaacttcga caagaacctg ccgaacgaaa     1560

aggtcctgcc gaagcacagc ctgctgtacg aatacttcac agtctacaac gaactgacaa     1620

aggtcaagta cgtcacagaa ggaatgagaa agccggcatt cctgagcgga gaacagaaga     1680

aggcaatcgt cgacctgctg ttcaagacaa acagaaaggt cacagtcaag cagctgaagg     1740

aagactactt caagaagatc gaatgcttcg acagcgtcga aatcagcgga gtcgaagaca     1800

gattcaacgc aagcctggga acataccacg acctgctgaa gatcatcaag gacaaggact     1860

tcctggacaa cgaagaaaac gaagacatcc tggaagacat cgtcctgaca ctgacactgt     1920

tcgaagacag agaaatgatc gaagaaagac tgaagacata cgcacacctg ttcgacgaca     1980

aggtcatgaa gcagctgaag agaagaagat acacaggatg gggaagactg agcagaaagc     2040

tgatcaacgg aatcagagac aagcagagcg gaaagacaat cctggacttc ctgaagagcg     2100

acggattcgc aaacagaaac ttcatgcagc tgatccacga cgacagcctg acattcaagg     2160

aagacatcca gaaggcacag gtcagcggac agggagacag cctgcacgaa cacatcgcaa     2220

acctggcagg aagcccggca atcaagaagg gaatcctgca gacagtcaag gtcgtcgacg     2280

aactggtcaa ggtcatggga agacacaagc cggaaaacat cgtcatcgaa atggcaagag     2340

aaaaccagac aacacagaag ggacagaaga acagcagaga aagaatgaag agaatcgaag     2400

aaggaatcaa ggaactggga agccagatcc tgaaggaaca cccggtcgaa aacacacagc     2460

tgcagaacga aaagctgtac ctgtactacc tgcagaacgg aagagacatg tacgtcgacc     2520

aggaactgga catcaacaga ctgagcgact acgacgtcga ccacatcgtc ccgcagagct     2580

tcctgaagga cgacagcatc gacaacaagg tcctgacaag aagcgacaag aacagaggaa     2640

agagcgacaa cgtcccgagc gaagaagtcg tcaagaagat gaagaactac tggagacagc     2700

tgctgaacgc aaagctgatc acacagagaa agttcgacaa cctgacaaag gcagagagag     2760

gaggactgag cgaactggac aaggcaggat tcatcaagag acagctggtc gaaacaagac     2820

agatcacaaa gcacgtcgca cagatcctgg acagcagaat gaacacaaag tacgacgaaa     2880

acgacaagct gatcagagaa gtcaaggtca tcacactgaa gagcaagctg gtcagcgact     2940

tcagaaagga cttccagttc tacaaggtca gagaaatcaa caactaccac cacgcacacg     3000

acgcatacct gaacgcagtc gtcggaacag cactgatcaa gaagtacccg aagctggaaa     3060

gcgaattcgt ctacggagac tacaaggtct acgacgtcag aaagatgatc gcaaagagcg     3120

aacaggaaat cggaaaggca acagcaaagt acttcttcta cagcaacatc atgaacttct     3180

tcaagacaga aatcacactg gcaaacggag aaatcagaaa gagaccgctg atcgaaacaa     3240

acggagaaac aggagaaatc gtctgggaca agggaagaga cttcgcaaca gtcagaaagg     3300

tcctgagcat gccgcaggtc aacatcgtca agaagacaga agtccagaca ggaggattca     3360

gcaaggaaag catcctgccg aagagaaaca gcgacaagct gatcgcaaga aagaaggact     3420

gggacccgaa gaagtacgga ggattcgaca gcccgacagt cgcatacagc gtcctggtcg     3480

tcgcaaaggt cgaaaaggga aagagcaaga agctgaagag cgtcaaggaa ctgctgggaa     3540

tcacaatcat ggaaagaagc agcttcgaaa agaacccgat cgacttcctg gaagcaaagg     3600

gatacaagga agtcaagaag gacctgatca tcaagctgcc gaagtacagc ctgttcgaac     3660

tggaaaacgg aagaaagaga atgctggcaa gcgcaggaga actgcagaag ggaaacgaac     3720

tggcactgcc gagcaagtac gtcaacttcc tgtacctggc aagccactac gaaaagctga     3780

agggaagccc ggaagacaac gaacagaagc agctgttcgt cgaacagcac aagcactacc     3840

tggacgaaat catcgaacag atcagcgaat tcagcaagag agtcatcctg gcagacgcaa     3900

acctggacaa ggtcctgagc gcatacaaca agcacagaga caagccgatc agagaacagg     3960

cagaaaacat catccacctg ttcacactga caaacctggg agcaccggca gcattcaagt     4020

acttcgacac aacaatcgac agaaagagat acacaagcac aaaggaagtc ctggacgcaa     4080

cactgatcca ccagagcatc acaggactgt acgaaacaag aatcgacctg agccagctgg     4140

gaggagacgg aggaggaagc ccgaagaaga agagaaaggt ctagctagca ccagcctcaa     4200

gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt     4260

cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattctc     4320

tcgag                                                                 4325


<210>  260
<211>  4325
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with AGG as first three nucleotides 
       for use with CleanCapTM, 5 UTR from XBG, ORF corresponding to SEQ
       ID NO: 204, Kozak sequence, and 3 UTR of XBG

<400>  260
aggaagctca gaataaacgc tcaactttgg ccggatctgc caccatggac aagaagtaca       60

gcatcggact ggacatcgga acaaacagcg tcggatgggc agtcatcaca gacgaataca      120

aggtcccgag caagaagttc aaggtcctgg gaaacacaga cagacacagc atcaagaaga      180

acctgatcgg agcactgctg ttcgacagcg gagaaacagc agaagcaaca agactgaaga      240

gaacagcaag aagaagatac acaagaagaa agaacagaat ctgctacctg caggaaatct      300

tcagcaacga aatggcaaag gtcgacgaca gcttcttcca cagactggaa gaaagcttcc      360

tggtcgaaga agacaagaag cacgaaagac acccgatctt cggaaacatc gtcgacgaag      420

tcgcatacca cgaaaagtac ccgacaatct accacctgag aaagaagctg gtcgacagca      480

cagacaaggc agacctgaga ctgatctacc tggcactggc acacatgatc aagttcagag      540

gacacttcct gatcgaagga gacctgaacc cggacaacag cgacgtcgac aagctgttca      600

tccagctggt ccagacatac aaccagctgt tcgaagaaaa cccgatcaac gcaagcggag      660

tcgacgcaaa ggcaatcctg agcgcaagac tgagcaagag cagaagactg gaaaacctga      720

tcgcacagct gccgggagaa aagaagaacg gactgttcgg aaacctgatc gcactgagcc      780

tgggactgac accgaacttc aagagcaact tcgacctggc agaagacgca aagctgcagc      840

tgagcaagga cacatacgac gacgacctgg acaacctgct ggcacagatc ggagaccagt      900

acgcagacct gttcctggca gcaaagaacc tgagcgacgc aatcctgctg agcgacatcc      960

tgagagtcaa cacagaaatc acaaaggcac cgctgagcgc aagcatgatc aagagatacg     1020

acgaacacca ccaggacctg acactgctga aggcactggt cagacagcag ctgccggaaa     1080

agtacaagga aatcttcttc gaccagagca agaacggata cgcaggatac atcgacggag     1140

gagcaagcca ggaagaattc tacaagttca tcaagccgat cctggaaaag atggacggaa     1200

cagaagaact gctggtcaag ctgaacagag aagacctgct gagaaagcag agaacattcg     1260

acaacggaag catcccgcac cagatccacc tgggagaact gcacgcaatc ctgagaagac     1320

aggaagactt ctacccgttc ctgaaggaca acagagaaaa gatcgaaaag atcctgacat     1380

tcagaatccc gtactacgtc ggaccgctgg caagaggaaa cagcagattc gcatggatga     1440

caagaaagag cgaagaaaca atcacaccgt ggaacttcga agaagtcgtc gacaagggag     1500

caagcgcaca gagcttcatc gaaagaatga caaacttcga caagaacctg ccgaacgaaa     1560

aggtcctgcc gaagcacagc ctgctgtacg aatacttcac agtctacaac gaactgacaa     1620

aggtcaagta cgtcacagaa ggaatgagaa agccggcatt cctgagcgga gaacagaaga     1680

aggcaatcgt cgacctgctg ttcaagacaa acagaaaggt cacagtcaag cagctgaagg     1740

aagactactt caagaagatc gaatgcttcg acagcgtcga aatcagcgga gtcgaagaca     1800

gattcaacgc aagcctggga acataccacg acctgctgaa gatcatcaag gacaaggact     1860

tcctggacaa cgaagaaaac gaagacatcc tggaagacat cgtcctgaca ctgacactgt     1920

tcgaagacag agaaatgatc gaagaaagac tgaagacata cgcacacctg ttcgacgaca     1980

aggtcatgaa gcagctgaag agaagaagat acacaggatg gggaagactg agcagaaagc     2040

tgatcaacgg aatcagagac aagcagagcg gaaagacaat cctggacttc ctgaagagcg     2100

acggattcgc aaacagaaac ttcatgcagc tgatccacga cgacagcctg acattcaagg     2160

aagacatcca gaaggcacag gtcagcggac agggagacag cctgcacgaa cacatcgcaa     2220

acctggcagg aagcccggca atcaagaagg gaatcctgca gacagtcaag gtcgtcgacg     2280

aactggtcaa ggtcatggga agacacaagc cggaaaacat cgtcatcgaa atggcaagag     2340

aaaaccagac aacacagaag ggacagaaga acagcagaga aagaatgaag agaatcgaag     2400

aaggaatcaa ggaactggga agccagatcc tgaaggaaca cccggtcgaa aacacacagc     2460

tgcagaacga aaagctgtac ctgtactacc tgcagaacgg aagagacatg tacgtcgacc     2520

aggaactgga catcaacaga ctgagcgact acgacgtcga ccacatcgtc ccgcagagct     2580

tcctgaagga cgacagcatc gacaacaagg tcctgacaag aagcgacaag aacagaggaa     2640

agagcgacaa cgtcccgagc gaagaagtcg tcaagaagat gaagaactac tggagacagc     2700

tgctgaacgc aaagctgatc acacagagaa agttcgacaa cctgacaaag gcagagagag     2760

gaggactgag cgaactggac aaggcaggat tcatcaagag acagctggtc gaaacaagac     2820

agatcacaaa gcacgtcgca cagatcctgg acagcagaat gaacacaaag tacgacgaaa     2880

acgacaagct gatcagagaa gtcaaggtca tcacactgaa gagcaagctg gtcagcgact     2940

tcagaaagga cttccagttc tacaaggtca gagaaatcaa caactaccac cacgcacacg     3000

acgcatacct gaacgcagtc gtcggaacag cactgatcaa gaagtacccg aagctggaaa     3060

gcgaattcgt ctacggagac tacaaggtct acgacgtcag aaagatgatc gcaaagagcg     3120

aacaggaaat cggaaaggca acagcaaagt acttcttcta cagcaacatc atgaacttct     3180

tcaagacaga aatcacactg gcaaacggag aaatcagaaa gagaccgctg atcgaaacaa     3240

acggagaaac aggagaaatc gtctgggaca agggaagaga cttcgcaaca gtcagaaagg     3300

tcctgagcat gccgcaggtc aacatcgtca agaagacaga agtccagaca ggaggattca     3360

gcaaggaaag catcctgccg aagagaaaca gcgacaagct gatcgcaaga aagaaggact     3420

gggacccgaa gaagtacgga ggattcgaca gcccgacagt cgcatacagc gtcctggtcg     3480

tcgcaaaggt cgaaaaggga aagagcaaga agctgaagag cgtcaaggaa ctgctgggaa     3540

tcacaatcat ggaaagaagc agcttcgaaa agaacccgat cgacttcctg gaagcaaagg     3600

gatacaagga agtcaagaag gacctgatca tcaagctgcc gaagtacagc ctgttcgaac     3660

tggaaaacgg aagaaagaga atgctggcaa gcgcaggaga actgcagaag ggaaacgaac     3720

tggcactgcc gagcaagtac gtcaacttcc tgtacctggc aagccactac gaaaagctga     3780

agggaagccc ggaagacaac gaacagaagc agctgttcgt cgaacagcac aagcactacc     3840

tggacgaaat catcgaacag atcagcgaat tcagcaagag agtcatcctg gcagacgcaa     3900

acctggacaa ggtcctgagc gcatacaaca agcacagaga caagccgatc agagaacagg     3960

cagaaaacat catccacctg ttcacactga caaacctggg agcaccggca gcattcaagt     4020

acttcgacac aacaatcgac agaaagagat acacaagcac aaaggaagtc ctggacgcaa     4080

cactgatcca ccagagcatc acaggactgt acgaaacaag aatcgacctg agccagctgg     4140

gaggagacgg aggaggaagc ccgaagaaga agagaaaggt ctagctagca ccagcctcaa     4200

gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt     4260

cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattctc     4320

tcgag                                                                 4325


<210>  261
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with AGG as first three nucleotides 
       for use with CleanCapTM,  5 UTR from HSD, ORF corresponding to 
       SEQ ID NO: 204, Kozak sequence, and 3 UTR of ALB

<400>  261
aggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  262

<400>  262
000

<210>  263
<211>  93
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: poly-A 100 sequence

<400>  263
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa       60

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa                                    93


<210>  264
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G209 single guide RNA targeting the mouse TTR gene

<400>  264
aaataagaga gaaaagaaga gtaagaagaa atataagagc cacc                        44


<210>  265
<211>  3312
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: ORF encoding Neisseria meningitidis Cas9 using minimal
       uridine codons, with start and stop codons

<400>  265
atggcagcat tcaagccgaa ctcgatcaac tacatcctgg gactggacat cggaatcgca       60

tcggtcggat gggcaatggt cgaaatcgac gaagaagaaa acccgatcag actgatcgac      120

ctgggagtca gagtcttcga aagagcagaa gtcccgaaga caggagactc gctggcaatg      180

gcaagaagac tggcaagatc ggtcagaaga ctgacaagaa gaagagcaca cagactgctg      240

agaacaagaa gactgctgaa gagagaagga gtcctgcagg cagcaaactt cgacgaaaac      300

ggactgatca agtcgctgcc gaacacaccg tggcagctga gagcagcagc actggacaga      360

aagctgacac cgctggaatg gtcggcagtc ctgctgcacc tgatcaagca cagaggatac      420

ctgtcgcaga gaaagaacga aggagaaaca gcagacaagg aactgggagc actgctgaag      480

ggagtcgcag gaaacgcaca cgcactgcag acaggagact tcagaacacc ggcagaactg      540

gcactgaaca agttcgaaaa ggaatcggga cacatcagaa accagagatc ggactactcg      600

cacacattct cgagaaagga cctgcaggca gaactgatcc tgctgttcga aaagcagaag      660

gaattcggaa acccgcacgt ctcgggagga ctgaaggaag gaatcgaaac actgctgatg      720

acacagagac cggcactgtc gggagacgca gtccagaaga tgctgggaca ctgcacattc      780

gaaccggcag aaccgaaggc agcaaagaac acatacacag cagaaagatt catctggctg      840

acaaagctga acaacctgag aatcctggaa cagggatcgg aaagaccgct gacagacaca      900

gaaagagcaa cactgatgga cgaaccgtac agaaagtcga agctgacata cgcacaggca      960

agaaagctgc tgggactgga agacacagca ttcttcaagg gactgagata cggaaaggac     1020

aacgcagaag catcgacact gatggaaatg aaggcatacc acgcaatctc gagagcactg     1080

gaaaaggaag gactgaagga caagaagtcg ccgctgaacc tgtcgccgga actgcaggac     1140

gaaatcggaa cagcattctc gctgttcaag acagacgaag acatcacagg aagactgaag     1200

gacagaatcc agccggaaat cctggaagca ctgctgaagc acatctcgtt cgacaagttc     1260

gtccagatct cgctgaaggc actgagaaga atcgtcccgc tgatggaaca gggaaagaga     1320

tacgacgaag catgcgcaga aatctacgga gaccactacg gaaagaagaa cacagaagaa     1380

aagatctacc tgccgccgat cccggcagac gaaatcagaa acccggtcgt cctgagagca     1440

ctgtcgcagg caagaaaggt catcaacgga gtcgtcagaa gatacggatc gccggcaaga     1500

atccacatcg aaacagcaag agaagtcgga aagtcgttca aggacagaaa ggaaatcgaa     1560

aagagacagg aagaaaacag aaaggacaga gaaaaggcag cagcaaagtt cagagaatac     1620

ttcccgaact tcgtcggaga accgaagtcg aaggacatcc tgaagctgag actgtacgaa     1680

cagcagcacg gaaagtgcct gtactcggga aaggaaatca acctgggaag actgaacgaa     1740

aagggatacg tcgaaatcga ccacgcactg ccgttctcga gaacatggga cgactcgttc     1800

aacaacaagg tcctggtcct gggatcggaa aaccagaaca agggaaacca gacaccgtac     1860

gaatacttca acggaaagga caactcgaga gaatggcagg aattcaaggc aagagtcgaa     1920

acatcgagat tcccgagatc gaagaagcag agaatcctgc tgcagaagtt cgacgaagac     1980

ggattcaagg aaagaaacct gaacgacaca agatacgtca acagattcct gtgccagttc     2040

gtcgcagaca gaatgagact gacaggaaag ggaaagaaga gagtcttcgc atcgaacgga     2100

cagatcacaa acctgctgag aggattctgg ggactgagaa aggtcagagc agaaaacgac     2160

agacaccacg cactggacgc agtcgtcgtc gcatgctcga cagtcgcaat gcagcagaag     2220

atcacaagat tcgtcagata caaggaaatg aacgcattcg acggaaagac aatcgacaag     2280

gaaacaggag aagtcctgca ccagaagaca cacttcccgc agccgtggga attcttcgca     2340

caggaagtca tgatcagagt cttcggaaag ccggacggaa agccggaatt cgaagaagca     2400

gacacactgg aaaagctgag aacactgctg gcagaaaagc tgtcgtcgag accggaagca     2460

gtccacgaat acgtcacacc gctgttcgtc tcgagagcac cgaacagaaa gatgtcggga     2520

cagggacaca tggaaacagt caagtcggca aagagactgg acgaaggagt ctcggtcctg     2580

agagtcccgc tgacacagct gaagctgaag gacctggaaa agatggtcaa cagagaaaga     2640

gaaccgaagc tgtacgaagc actgaaggca agactggaag cacacaagga cgacccggca     2700

aaggcattcg cagaaccgtt ctacaagtac gacaaggcag gaaacagaac acagcaggtc     2760

aaggcagtca gagtcgaaca ggtccagaag acaggagtct gggtcagaaa ccacaacgga     2820

atcgcagaca acgcaacaat ggtcagagta gacgtcttcg aaaagggaga caagtactac     2880

ctggtcccga tctactcgtg gcaggtcgca aagggaatcc tgccggacag agcagtcgtc     2940

cagggaaagg acgaagaaga ctggcagctg atcgacgact cgttcaactt caagttctcg     3000

ctgcacccga acgacctggt cgaagtcatc acaaagaagg caagaatgtt cggatacttc     3060

gcatcgtgcc acagaggaac aggaaacatc aacatcagaa tccacgacct ggaccacaag     3120

atcggaaaga acggaatcct ggaaggaatc ggagtcaaga cagcactgtc gttccagaag     3180

taccagatcg acgaactggg aaaggaaatc agaccgtgca gactgaagaa gagaccgccg     3240

gtcagatccg gaaagagaac agcagacgga tcggaattcg aatcgccgaa gaagaagaga     3300

aaggtcgaat ga                                                         3312


<210>  266
<211>  3306
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: ORF encoding Neisseria meningitidis Cas9 using minimal
       uridine codons (no start or stop codons; suitable for inclusion 
       in fusion protein coding sequence)

<400>  266
gcagcattca agccgaactc gatcaactac atcctgggac tggacatcgg aatcgcatcg       60

gtcggatggg caatggtcga aatcgacgaa gaagaaaacc cgatcagact gatcgacctg      120

ggagtcagag tcttcgaaag agcagaagtc ccgaagacag gagactcgct ggcaatggca      180

agaagactgg caagatcggt cagaagactg acaagaagaa gagcacacag actgctgaga      240

acaagaagac tgctgaagag agaaggagtc ctgcaggcag caaacttcga cgaaaacgga      300

ctgatcaagt cgctgccgaa cacaccgtgg cagctgagag cagcagcact ggacagaaag      360

ctgacaccgc tggaatggtc ggcagtcctg ctgcacctga tcaagcacag aggatacctg      420

tcgcagagaa agaacgaagg agaaacagca gacaaggaac tgggagcact gctgaaggga      480

gtcgcaggaa acgcacacgc actgcagaca ggagacttca gaacaccggc agaactggca      540

ctgaacaagt tcgaaaagga atcgggacac atcagaaacc agagatcgga ctactcgcac      600

acattctcga gaaaggacct gcaggcagaa ctgatcctgc tgttcgaaaa gcagaaggaa      660

ttcggaaacc cgcacgtctc gggaggactg aaggaaggaa tcgaaacact gctgatgaca      720

cagagaccgg cactgtcggg agacgcagtc cagaagatgc tgggacactg cacattcgaa      780

ccggcagaac cgaaggcagc aaagaacaca tacacagcag aaagattcat ctggctgaca      840

aagctgaaca acctgagaat cctggaacag ggatcggaaa gaccgctgac agacacagaa      900

agagcaacac tgatggacga accgtacaga aagtcgaagc tgacatacgc acaggcaaga      960

aagctgctgg gactggaaga cacagcattc ttcaagggac tgagatacgg aaaggacaac     1020

gcagaagcat cgacactgat ggaaatgaag gcataccacg caatctcgag agcactggaa     1080

aaggaaggac tgaaggacaa gaagtcgccg ctgaacctgt cgccggaact gcaggacgaa     1140

atcggaacag cattctcgct gttcaagaca gacgaagaca tcacaggaag actgaaggac     1200

agaatccagc cggaaatcct ggaagcactg ctgaagcaca tctcgttcga caagttcgtc     1260

cagatctcgc tgaaggcact gagaagaatc gtcccgctga tggaacaggg aaagagatac     1320

gacgaagcat gcgcagaaat ctacggagac cactacggaa agaagaacac agaagaaaag     1380

atctacctgc cgccgatccc ggcagacgaa atcagaaacc cggtcgtcct gagagcactg     1440

tcgcaggcaa gaaaggtcat caacggagtc gtcagaagat acggatcgcc ggcaagaatc     1500

cacatcgaaa cagcaagaga agtcggaaag tcgttcaagg acagaaagga aatcgaaaag     1560

agacaggaag aaaacagaaa ggacagagaa aaggcagcag caaagttcag agaatacttc     1620

ccgaacttcg tcggagaacc gaagtcgaag gacatcctga agctgagact gtacgaacag     1680

cagcacggaa agtgcctgta ctcgggaaag gaaatcaacc tgggaagact gaacgaaaag     1740

ggatacgtcg aaatcgacca cgcactgccg ttctcgagaa catgggacga ctcgttcaac     1800

aacaaggtcc tggtcctggg atcggaaaac cagaacaagg gaaaccagac accgtacgaa     1860

tacttcaacg gaaaggacaa ctcgagagaa tggcaggaat tcaaggcaag agtcgaaaca     1920

tcgagattcc cgagatcgaa gaagcagaga atcctgctgc agaagttcga cgaagacgga     1980

ttcaaggaaa gaaacctgaa cgacacaaga tacgtcaaca gattcctgtg ccagttcgtc     2040

gcagacagaa tgagactgac aggaaaggga aagaagagag tcttcgcatc gaacggacag     2100

atcacaaacc tgctgagagg attctgggga ctgagaaagg tcagagcaga aaacgacaga     2160

caccacgcac tggacgcagt cgtcgtcgca tgctcgacag tcgcaatgca gcagaagatc     2220

acaagattcg tcagatacaa ggaaatgaac gcattcgacg gaaagacaat cgacaaggaa     2280

acaggagaag tcctgcacca gaagacacac ttcccgcagc cgtgggaatt cttcgcacag     2340

gaagtcatga tcagagtctt cggaaagccg gacggaaagc cggaattcga agaagcagac     2400

acactggaaa agctgagaac actgctggca gaaaagctgt cgtcgagacc ggaagcagtc     2460

cacgaatacg tcacaccgct gttcgtctcg agagcaccga acagaaagat gtcgggacag     2520

ggacacatgg aaacagtcaa gtcggcaaag agactggacg aaggagtctc ggtcctgaga     2580

gtcccgctga cacagctgaa gctgaaggac ctggaaaaga tggtcaacag agaaagagaa     2640

ccgaagctgt acgaagcact gaaggcaaga ctggaagcac acaaggacga cccggcaaag     2700

gcattcgcag aaccgttcta caagtacgac aaggcaggaa acagaacaca gcaggtcaag     2760

gcagtcagag tcgaacaggt ccagaagaca ggagtctggg tcagaaacca caacggaatc     2820

gcagacaacg caacaatggt cagagtagac gtcttcgaaa agggagacaa gtactacctg     2880

gtcccgatct actcgtggca ggtcgcaaag ggaatcctgc cggacagagc agtcgtccag     2940

ggaaaggacg aagaagactg gcagctgatc gacgactcgt tcaacttcaa gttctcgctg     3000

cacccgaacg acctggtcga agtcatcaca aagaaggcaa gaatgttcgg atacttcgca     3060

tcgtgccaca gaggaacagg aaacatcaac atcagaatcc acgacctgga ccacaagatc     3120

ggaaagaacg gaatcctgga aggaatcgga gtcaagacag cactgtcgtt ccagaagtac     3180

cagatcgacg aactgggaaa ggaaatcaga ccgtgcagac tgaagaagag accgccggtc     3240

agatccggaa agagaacagc agacggatcg gaattcgaat cgccgaagaa gaagagaaag     3300

gtcgaa                                                                3306


<210>  267
<211>  3636
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Transcript comprising SEQ ID NO: 265 (encoding 
       Neisseria meningitidis Cas9)

<400>  267
gggagaccca agctggctag cgtttaaact taagcttgga tccgccacca tggcagcatt       60

caagccgaac tcgatcaact acatcctggg actggacatc ggaatcgcat cggtcggatg      120

ggcaatggtc gaaatcgacg aagaagaaaa cccgatcaga ctgatcgacc tgggagtcag      180

agtcttcgaa agagcagaag tcccgaagac aggagactcg ctggcaatgg caagaagact      240

ggcaagatcg gtcagaagac tgacaagaag aagagcacac agactgctga gaacaagaag      300

actgctgaag agagaaggag tcctgcaggc agcaaacttc gacgaaaacg gactgatcaa      360

gtcgctgccg aacacaccgt ggcagctgag agcagcagca ctggacagaa agctgacacc      420

gctggaatgg tcggcagtcc tgctgcacct gatcaagcac agaggatacc tgtcgcagag      480

aaagaacgaa ggagaaacag cagacaagga actgggagca ctgctgaagg gagtcgcagg      540

aaacgcacac gcactgcaga caggagactt cagaacaccg gcagaactgg cactgaacaa      600

gttcgaaaag gaatcgggac acatcagaaa ccagagatcg gactactcgc acacattctc      660

gagaaaggac ctgcaggcag aactgatcct gctgttcgaa aagcagaagg aattcggaaa      720

cccgcacgtc tcgggaggac tgaaggaagg aatcgaaaca ctgctgatga cacagagacc      780

ggcactgtcg ggagacgcag tccagaagat gctgggacac tgcacattcg aaccggcaga      840

accgaaggca gcaaagaaca catacacagc agaaagattc atctggctga caaagctgaa      900

caacctgaga atcctggaac agggatcgga aagaccgctg acagacacag aaagagcaac      960

actgatggac gaaccgtaca gaaagtcgaa gctgacatac gcacaggcaa gaaagctgct     1020

gggactggaa gacacagcat tcttcaaggg actgagatac ggaaaggaca acgcagaagc     1080

atcgacactg atggaaatga aggcatacca cgcaatctcg agagcactgg aaaaggaagg     1140

actgaaggac aagaagtcgc cgctgaacct gtcgccggaa ctgcaggacg aaatcggaac     1200

agcattctcg ctgttcaaga cagacgaaga catcacagga agactgaagg acagaatcca     1260

gccggaaatc ctggaagcac tgctgaagca catctcgttc gacaagttcg tccagatctc     1320

gctgaaggca ctgagaagaa tcgtcccgct gatggaacag ggaaagagat acgacgaagc     1380

atgcgcagaa atctacggag accactacgg aaagaagaac acagaagaaa agatctacct     1440

gccgccgatc ccggcagacg aaatcagaaa cccggtcgtc ctgagagcac tgtcgcaggc     1500

aagaaaggtc atcaacggag tcgtcagaag atacggatcg ccggcaagaa tccacatcga     1560

aacagcaaga gaagtcggaa agtcgttcaa ggacagaaag gaaatcgaaa agagacagga     1620

agaaaacaga aaggacagag aaaaggcagc agcaaagttc agagaatact tcccgaactt     1680

cgtcggagaa ccgaagtcga aggacatcct gaagctgaga ctgtacgaac agcagcacgg     1740

aaagtgcctg tactcgggaa aggaaatcaa cctgggaaga ctgaacgaaa agggatacgt     1800

cgaaatcgac cacgcactgc cgttctcgag aacatgggac gactcgttca acaacaaggt     1860

cctggtcctg ggatcggaaa accagaacaa gggaaaccag acaccgtacg aatacttcaa     1920

cggaaaggac aactcgagag aatggcagga attcaaggca agagtcgaaa catcgagatt     1980

cccgagatcg aagaagcaga gaatcctgct gcagaagttc gacgaagacg gattcaagga     2040

aagaaacctg aacgacacaa gatacgtcaa cagattcctg tgccagttcg tcgcagacag     2100

aatgagactg acaggaaagg gaaagaagag agtcttcgca tcgaacggac agatcacaaa     2160

cctgctgaga ggattctggg gactgagaaa ggtcagagca gaaaacgaca gacaccacgc     2220

actggacgca gtcgtcgtcg catgctcgac agtcgcaatg cagcagaaga tcacaagatt     2280

cgtcagatac aaggaaatga acgcattcga cggaaagaca atcgacaagg aaacaggaga     2340

agtcctgcac cagaagacac acttcccgca gccgtgggaa ttcttcgcac aggaagtcat     2400

gatcagagtc ttcggaaagc cggacggaaa gccggaattc gaagaagcag acacactgga     2460

aaagctgaga acactgctgg cagaaaagct gtcgtcgaga ccggaagcag tccacgaata     2520

cgtcacaccg ctgttcgtct cgagagcacc gaacagaaag atgtcgggac agggacacat     2580

ggaaacagtc aagtcggcaa agagactgga cgaaggagtc tcggtcctga gagtcccgct     2640

gacacagctg aagctgaagg acctggaaaa gatggtcaac agagaaagag aaccgaagct     2700

gtacgaagca ctgaaggcaa gactggaagc acacaaggac gacccggcaa aggcattcgc     2760

agaaccgttc tacaagtacg acaaggcagg aaacagaaca cagcaggtca aggcagtcag     2820

agtcgaacag gtccagaaga caggagtctg ggtcagaaac cacaacggaa tcgcagacaa     2880

cgcaacaatg gtcagagtag acgtcttcga aaagggagac aagtactacc tggtcccgat     2940

ctactcgtgg caggtcgcaa agggaatcct gccggacaga gcagtcgtcc agggaaagga     3000

cgaagaagac tggcagctga tcgacgactc gttcaacttc aagttctcgc tgcacccgaa     3060

cgacctggtc gaagtcatca caaagaaggc aagaatgttc ggatacttcg catcgtgcca     3120

cagaggaaca ggaaacatca acatcagaat ccacgacctg gaccacaaga tcggaaagaa     3180

cggaatcctg gaaggaatcg gagtcaagac agcactgtcg ttccagaagt accagatcga     3240

cgaactggga aaggaaatca gaccgtgcag actgaagaag agaccgccgg tcagatccgg     3300

aaagagaaca gcagacggat cggaattcga atcgccgaag aagaagagaa aggtcgaatg     3360

atagctagct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt     3420

ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg     3480

ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt     3540

gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca     3600

atagcaggca tgctggggat gcggtgggct ctatgg                               3636


<210>  268
<211>  1103
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Neisseria meningitidis Cas9

<400>  268

Met Ala Ala Phe Lys Pro Asn Ser Ile Asn Tyr Ile Leu Gly Leu Asp 
1               5                   10                  15      


Ile Gly Ile Ala Ser Val Gly Trp Ala Met Val Glu Ile Asp Glu Glu 
            20                  25                  30          


Glu Asn Pro Ile Arg Leu Ile Asp Leu Gly Val Arg Val Phe Glu Arg 
        35                  40                  45              


Ala Glu Val Pro Lys Thr Gly Asp Ser Leu Ala Met Ala Arg Arg Leu 
    50                  55                  60                  


Ala Arg Ser Val Arg Arg Leu Thr Arg Arg Arg Ala His Arg Leu Leu 
65                  70                  75                  80  


Arg Thr Arg Arg Leu Leu Lys Arg Glu Gly Val Leu Gln Ala Ala Asn 
                85                  90                  95      


Phe Asp Glu Asn Gly Leu Ile Lys Ser Leu Pro Asn Thr Pro Trp Gln 
            100                 105                 110         


Leu Arg Ala Ala Ala Leu Asp Arg Lys Leu Thr Pro Leu Glu Trp Ser 
        115                 120                 125             


Ala Val Leu Leu His Leu Ile Lys His Arg Gly Tyr Leu Ser Gln Arg 
    130                 135                 140                 


Lys Asn Glu Gly Glu Thr Ala Asp Lys Glu Leu Gly Ala Leu Leu Lys 
145                 150                 155                 160 


Gly Val Ala Gly Asn Ala His Ala Leu Gln Thr Gly Asp Phe Arg Thr 
                165                 170                 175     


Pro Ala Glu Leu Ala Leu Asn Lys Phe Glu Lys Glu Ser Gly His Ile 
            180                 185                 190         


Arg Asn Gln Arg Ser Asp Tyr Ser His Thr Phe Ser Arg Lys Asp Leu 
        195                 200                 205             


Gln Ala Glu Leu Ile Leu Leu Phe Glu Lys Gln Lys Glu Phe Gly Asn 
    210                 215                 220                 


Pro His Val Ser Gly Gly Leu Lys Glu Gly Ile Glu Thr Leu Leu Met 
225                 230                 235                 240 


Thr Gln Arg Pro Ala Leu Ser Gly Asp Ala Val Gln Lys Met Leu Gly 
                245                 250                 255     


His Cys Thr Phe Glu Pro Ala Glu Pro Lys Ala Ala Lys Asn Thr Tyr 
            260                 265                 270         


Thr Ala Glu Arg Phe Ile Trp Leu Thr Lys Leu Asn Asn Leu Arg Ile 
        275                 280                 285             


Leu Glu Gln Gly Ser Glu Arg Pro Leu Thr Asp Thr Glu Arg Ala Thr 
    290                 295                 300                 


Leu Met Asp Glu Pro Tyr Arg Lys Ser Lys Leu Thr Tyr Ala Gln Ala 
305                 310                 315                 320 


Arg Lys Leu Leu Gly Leu Glu Asp Thr Ala Phe Phe Lys Gly Leu Arg 
                325                 330                 335     


Tyr Gly Lys Asp Asn Ala Glu Ala Ser Thr Leu Met Glu Met Lys Ala 
            340                 345                 350         


Tyr His Ala Ile Ser Arg Ala Leu Glu Lys Glu Gly Leu Lys Asp Lys 
        355                 360                 365             


Lys Ser Pro Leu Asn Leu Ser Pro Glu Leu Gln Asp Glu Ile Gly Thr 
    370                 375                 380                 


Ala Phe Ser Leu Phe Lys Thr Asp Glu Asp Ile Thr Gly Arg Leu Lys 
385                 390                 395                 400 


Asp Arg Ile Gln Pro Glu Ile Leu Glu Ala Leu Leu Lys His Ile Ser 
                405                 410                 415     


Phe Asp Lys Phe Val Gln Ile Ser Leu Lys Ala Leu Arg Arg Ile Val 
            420                 425                 430         


Pro Leu Met Glu Gln Gly Lys Arg Tyr Asp Glu Ala Cys Ala Glu Ile 
        435                 440                 445             


Tyr Gly Asp His Tyr Gly Lys Lys Asn Thr Glu Glu Lys Ile Tyr Leu 
    450                 455                 460                 


Pro Pro Ile Pro Ala Asp Glu Ile Arg Asn Pro Val Val Leu Arg Ala 
465                 470                 475                 480 


Leu Ser Gln Ala Arg Lys Val Ile Asn Gly Val Val Arg Arg Tyr Gly 
                485                 490                 495     


Ser Pro Ala Arg Ile His Ile Glu Thr Ala Arg Glu Val Gly Lys Ser 
            500                 505                 510         


Phe Lys Asp Arg Lys Glu Ile Glu Lys Arg Gln Glu Glu Asn Arg Lys 
        515                 520                 525             


Asp Arg Glu Lys Ala Ala Ala Lys Phe Arg Glu Tyr Phe Pro Asn Phe 
    530                 535                 540                 


Val Gly Glu Pro Lys Ser Lys Asp Ile Leu Lys Leu Arg Leu Tyr Glu 
545                 550                 555                 560 


Gln Gln His Gly Lys Cys Leu Tyr Ser Gly Lys Glu Ile Asn Leu Gly 
                565                 570                 575     


Arg Leu Asn Glu Lys Gly Tyr Val Glu Ile Asp His Ala Leu Pro Phe 
            580                 585                 590         


Ser Arg Thr Trp Asp Asp Ser Phe Asn Asn Lys Val Leu Val Leu Gly 
        595                 600                 605             


Ser Glu Asn Gln Asn Lys Gly Asn Gln Thr Pro Tyr Glu Tyr Phe Asn 
    610                 615                 620                 


Gly Lys Asp Asn Ser Arg Glu Trp Gln Glu Phe Lys Ala Arg Val Glu 
625                 630                 635                 640 


Thr Ser Arg Phe Pro Arg Ser Lys Lys Gln Arg Ile Leu Leu Gln Lys 
                645                 650                 655     


Phe Asp Glu Asp Gly Phe Lys Glu Arg Asn Leu Asn Asp Thr Arg Tyr 
            660                 665                 670         


Val Asn Arg Phe Leu Cys Gln Phe Val Ala Asp Arg Met Arg Leu Thr 
        675                 680                 685             


Gly Lys Gly Lys Lys Arg Val Phe Ala Ser Asn Gly Gln Ile Thr Asn 
    690                 695                 700                 


Leu Leu Arg Gly Phe Trp Gly Leu Arg Lys Val Arg Ala Glu Asn Asp 
705                 710                 715                 720 


Arg His His Ala Leu Asp Ala Val Val Val Ala Cys Ser Thr Val Ala 
                725                 730                 735     


Met Gln Gln Lys Ile Thr Arg Phe Val Arg Tyr Lys Glu Met Asn Ala 
            740                 745                 750         


Phe Asp Gly Lys Thr Ile Asp Lys Glu Thr Gly Glu Val Leu His Gln 
        755                 760                 765             


Lys Thr His Phe Pro Gln Pro Trp Glu Phe Phe Ala Gln Glu Val Met 
    770                 775                 780                 


Ile Arg Val Phe Gly Lys Pro Asp Gly Lys Pro Glu Phe Glu Glu Ala 
785                 790                 795                 800 


Asp Thr Leu Glu Lys Leu Arg Thr Leu Leu Ala Glu Lys Leu Ser Ser 
                805                 810                 815     


Arg Pro Glu Ala Val His Glu Tyr Val Thr Pro Leu Phe Val Ser Arg 
            820                 825                 830         


Ala Pro Asn Arg Lys Met Ser Gly Gln Gly His Met Glu Thr Val Lys 
        835                 840                 845             


Ser Ala Lys Arg Leu Asp Glu Gly Val Ser Val Leu Arg Val Pro Leu 
    850                 855                 860                 


Thr Gln Leu Lys Leu Lys Asp Leu Glu Lys Met Val Asn Arg Glu Arg 
865                 870                 875                 880 


Glu Pro Lys Leu Tyr Glu Ala Leu Lys Ala Arg Leu Glu Ala His Lys 
                885                 890                 895     


Asp Asp Pro Ala Lys Ala Phe Ala Glu Pro Phe Tyr Lys Tyr Asp Lys 
            900                 905                 910         


Ala Gly Asn Arg Thr Gln Gln Val Lys Ala Val Arg Val Glu Gln Val 
        915                 920                 925             


Gln Lys Thr Gly Val Trp Val Arg Asn His Asn Gly Ile Ala Asp Asn 
    930                 935                 940                 


Ala Thr Met Val Arg Val Asp Val Phe Glu Lys Gly Asp Lys Tyr Tyr 
945                 950                 955                 960 


Leu Val Pro Ile Tyr Ser Trp Gln Val Ala Lys Gly Ile Leu Pro Asp 
                965                 970                 975     


Arg Ala Val Val Gln Gly Lys Asp Glu Glu Asp Trp Gln Leu Ile Asp 
            980                 985                 990         


Asp Ser Phe Asn Phe Lys Phe Ser  Leu His Pro Asn Asp  Leu Val Glu 
        995                 1000                 1005             


Val Ile  Thr Lys Lys Ala Arg  Met Phe Gly Tyr Phe  Ala Ser Cys 
    1010                 1015                 1020             


His Arg  Gly Thr Gly Asn Ile  Asn Ile Arg Ile His  Asp Leu Asp 
    1025                 1030                 1035             


His Lys  Ile Gly Lys Asn Gly  Ile Leu Glu Gly Ile  Gly Val Lys 
    1040                 1045                 1050             


Thr Ala  Leu Ser Phe Gln Lys  Tyr Gln Ile Asp Glu  Leu Gly Lys 
    1055                 1060                 1065             


Glu Ile  Arg Pro Cys Arg Leu  Lys Lys Arg Pro Pro  Val Arg Ser 
    1070                 1075                 1080             


Gly Lys  Arg Thr Ala Asp Gly  Ser Glu Phe Glu Ser  Pro Lys Lys 
    1085                 1090                 1095             


Lys Arg  Lys Val Glu 
    1100             


<210>  269
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G390 single guide RNA targeting the rat TTR gene


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  269
gccgagucug gagagcugca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  270
<211>  74
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: trRNA

<400>  270
aacagcauag caaguuaaaa uaaggcuagu ccguuaucaa cuugaaaaag uggcaccgag       60

ucggugcuuu uuuu                                                         74


<210>  271

<400>  271
000

<210>  272
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G534 single guide RNA targeting the rat TTR gene


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  272
acgcaaauau caguccagcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  273
<211>  95
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G000395 5 truncated inactive sgRNA modified sequence


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (24)..(35)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (64)..(91)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (92)..(95)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  273
gcaauggugu agcggguuuu agagcuagaa auagcaaguu aaaauaaggc uaguccguua       60

ucaacuugaa aaaguggcac cgagucggug cuuuu                                  95


<210>  274
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: SV40 NLS

<400>  274

Pro Lys Lys Lys Arg Lys Val 
1               5           


<210>  275
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Alternate SV40 NLS

<400>  275

Pro Lys Lys Lys Arg Arg Val 
1               5           


<210>  276
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Nucleoplasmin NLS

<400>  276

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys 
1               5                   10                  15      


<210>  277
<211>  10
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary Kozak sequence

<400>  277
gccrccaugg                                                              10


<210>  278
<211>  13
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary Kozak sequence

<400>  278
gccgccrcca ugg                                                          13


