                         SEQUENCE LISTING

<110>  LightBio Limited
 
<120>  Nucleic Acid Construct

<130>  P32190WO1

<150>  1913898.1
<151>  2019-09-26

<160>  7     

<170>  PatentIn version 3.5

<210>  1
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PAM sequence

<400>  1
gtcaccaatc ctgtccctag tgg                                               23


<210>  2
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  sgRNA guide sequence

<400>  2
gucaccaauc cugucccuag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  3
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PAM sequence 2

<400>  3
gctgaagcac tgcacgccgt agg                                               23


<210>  4
<211>  140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HDR template

<400>  4
accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc       60

accctgagcc acggggtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac      120

ttcttcaagt ccgccatgcc                                                  140


<210>  5
<211>  720
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EGFP sequence

<400>  5
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac       60

ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac      120

ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc      180

ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag      240

cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc      300

ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg      360

gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac      420

aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac      480

ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc      540

gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac      600

tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc      660

ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag      720


<210>  6
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  sgRNA guide sequence 2

<400>  6
gcugaagcac ugcacgccgu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  7
<211>  9177
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Cas9/sgRNA plasmid sequence

<400>  7
gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag       60

ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga      120

aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat      180

atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga      240

cgaaacaccg gctgaagcac tgcacgccgt gttttagagc tagaaatagc aagttaaaat      300

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga      360

gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca      420

gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac      480

cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac      540

gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata      600

tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc      660

agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta      720

ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc      780

cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg      840

gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc      900

ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga      960

ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc     1020

gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac     1080

tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt     1140

agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt     1200

acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggacta     1260

taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa     1320

gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa     1380

gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga     1440

gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa     1500

gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct     1560

gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga     1620

gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc     1680

cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga     1740

cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga     1800

cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt     1860

ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct     1920

gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag     1980

cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa     2040

tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct     2100

gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact     2160

gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga     2220

ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga     2280

catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag     2340

atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc     2400

tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga     2460

cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga     2520

cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac     2580

cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg     2640

gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct     2700

gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg     2760

gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa     2820

gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa     2880

cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct     2940

gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca     3000

gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct     3060

gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga     3120

agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa     3180

ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac     3240

actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga     3300

cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg     3360

gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa     3420

gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt     3480

taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat     3540

tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt     3600

ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc     3660

cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat     3720

cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac     3780

ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt     3840

ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca     3900

gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg     3960

gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg     4020

gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga     4080

gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac     4140

ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga     4200

cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc     4260

cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc     4320

ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct     4380

ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa     4440

gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa     4500

ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga     4560

gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg     4620

gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg     4680

cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa     4740

ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct     4800

ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct     4860

ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc     4920

caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt     4980

cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa     5040

cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa     5100

gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca     5160

ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga     5220

cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga     5280

gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt     5340

caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga     5400

cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca     5460

gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa     5520

ggaattcggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa     5580

tcctggccca atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc     5640

cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt     5700

cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt     5760

cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac     5820

cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga     5880

gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg     5940

gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa     6000

gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc     6060

cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac     6120

cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc     6180

cggtgcctga gaattctaac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc     6240

cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc     6300

actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct     6360

attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg     6420

catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc     6480

gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg     6540

gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt     6600

ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc     6660

cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac     6720

ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg     6780

ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt     6840

tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc     6900

cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct     6960

tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga     7020

ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga     7080

attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg     7140

atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg     7200

cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt     7260

gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc     7320

tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc     7380

ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc     7440

cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga     7500

gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt     7560

ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag     7620

tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag     7680

aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta     7740

ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg     7800

agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca     7860

gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag     7920

gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc     7980

gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg     8040

tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc     8100

ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg     8160

cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg     8220

gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga     8280

cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac     8340

tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa     8400

aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca     8460

aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag     8520

gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac     8580

cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa     8640

ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc     8700

accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag     8760

tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac     8820

cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc     8880

gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc     8940

ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca     9000

cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc     9060

tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg     9120

ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt        9177


