                         SEQUENCE LISTING

<110>  Children's Hospital Medical Center
 
<120>  MODEL FOR INSULIN RESISTANCE

<130>  CHMC63.032WO

<150>  US 63/042,997
<151>  2020-06-23

<160>  18    

<170>  PatentIn version 3.5

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

<220>
<223>  Human PCK1 sgRNA 1

<400>  1
gaataagcca gatgtaatca ggg                                               23


<210>  2
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Human PCK1 sgRNA 2

<400>  2
actcaggccc tgattacatc tgg                                               23


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

<220>
<223>  Human PCK1 sgRNA 3

<400>  3
tctctctctc gatttcacag ggg                                               23


<210>  4
<211>  7024
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  hPCK1 2A-mSc-2A-Luc


<220>
<221>  misc_feature
<222>  (1)..(1561)
<223>  5' PCK1 homology

<220>
<221>  misc_feature
<222>  (1565)..(1630)
<223>  P2A self-cleaving peptide

<220>
<221>  misc_feature
<222>  (1631)..(2326)
<223>  mScarlet

<220>
<221>  misc_feature
<222>  (2327)..(2389)
<223>  T2A self-cleaving peptide

<220>
<221>  misc_feature
<222>  (2390)..(4042)
<223>  Luciferase

<220>
<221>  misc_feature
<222>  (4043)..(4076)
<223>  LoxP

<220>
<221>  misc_feature
<222>  (4077)..(5319)
<223>  NeoR

<220>
<221>  misc_feature
<222>  (5320)..(5353)
<223>  LoxP

<220>
<221>  misc_feature
<222>  (5379)..(7024)
<223>  3' PCK1 homology

<400>  4
ccaatgccat caagaccatc cagaagaaca caatctttac caatgtggcc gagaccagcg       60

acgggggcgt ttactgggaa ggcattgatg agccgctagc ttcaggtgtc accatcacgt      120

cctggaagaa taaggagtgg agctcagagg atggtgtgtc cctgccagag gccttggtgt      180

gccgggctgc agggactgcc tgtttgagcc aggcactcac gagcctttct ctgtcttata      240

ggggaacctt gtgcccaccc caactcgagg ttctgcaccc ctgccagcca gtgccccatc      300

attgatgctg cctgggagtc tccggaaggt gttcccattg aaggcattat ctttggaggc      360

cgtagacctg ctggtgaggc tctccttcat ttaggctggg aacatgggtg tgctgggtac      420

cgaagggcat ctgtgaaatc tctccttttc catgaccttg tcagagggtg cccaggggct      480

tcctttcttg agcttccttc ccaaagatcc agaataattg gcaagttcaa atgtagaacc      540

aacccttctg gtcacctgga acctttctga atccctgatc tattgtagct tgatcaaatt      600

ttacttttta ctttgtggcc tcagtcatgt aactttgagt tagcagtttt ctgcaattta      660

gcttggtgaa cgcaaaacta gctcgattac aaagttattg tcttgccgtg tctttccgtg      720

ttgtgaataa caccactggt tgtggagtct gaatttcaaa gcctctgatg aacatttctc      780

tttttttttc ctgctaaagg tgtccctcta gtctatgaag ctctcagctg gcaacatgga      840

gtctttgtgg gggcggccat gagatcagag gccacagcgg ctgcagaaca taaaggtaaa      900

tcaaagtcct gatctgaaac cacagagaag tgggattaga gcactcttcg tcactcttat      960

gtctctctcc ttttctgtgt ctgtgtgtgg ggagagagag agagagaaag agagagagga     1020

gaacaaagca tgctaatgtc aacaatcaat ggcgtcagtc ttgcctagga gagcctcatt     1080

tactaatgaa ctccctctct gtttaacagg caaaatcatc atgcatgacc cctttgccat     1140

gcggcccttc tttggctaca acttcggcaa atacctggcc cactggctta gcatggccca     1200

gcacccagca gccaaactgc ccaagatctt ccatgtcaac tggttccgga aggacaagga     1260

aggcaaattc ctctggccag gctttggaga gaactccagg gtgctggagt ggatgttcaa     1320

ccggatcgat ggaaaagcca gcaccaagct cacgcccata ggctacatcc ccaaggagga     1380

tgccctgaac ctgaaaggcc tggggcacat caacatgatg gagcttttca gcatctccaa     1440

ggaattctgg gagaaggagg tggaagacat cgagaagtat ctggaggatc aagtcaatgc     1500

cgacctcccc tgtgaaatcg agagagagat ccttgccttg aagcaaagaa taagccagat     1560

ggggggatcc ggagctacta acttcagcct gctgaagcag gctggagatg tggaggagaa     1620

ccctggacct atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt     1680

gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg     1740

cccctacgag ggcacccaga ccgccaagct gaaggtgaca aagggtggcc ccctgccctt     1800

ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc     1860

cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt     1920

gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg     1980

caccctgatc tacaaggtga agctgcgcgg caccaacttc cctcctgacg gccccgtaat     2040

gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt     2100

gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcgggcgct acctggcgga     2160

cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtgga     2220

ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg     2280

ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggca gcggagaggg     2340

cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta tggaagacgc     2400

caaaaacata aagaaaggcc cggcgccatt ctatccgcta gaggatggaa ccgctggaga     2460

gcaactgcat aaggctatga agagatacgc cctggttcct ggaaccattg cttttacaga     2520

tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg ttcggttggc     2580

agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa     2640

ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc     2700

cgcgaacgac atctataatg aacgtgaatt gctcaacagt atgaacattt cgcagcctac     2760

cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaaattacc     2820

aataatccag aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat     2880

gtacaccttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtaccaga     2940

gtcctttgat cgtgacaaaa caattgcact gataatgaac tcctctggat ctactgggtt     3000

acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc atgccagaga     3060

tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca     3120

tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt     3180

aatgtataga tttgaagagg agctgttttt acgatccctt caggattaca aaatccaaag     3240

tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga ttgacaaata     3300

cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga aagaagtcgg     3360

ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg ggctcactga     3420

gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa     3480

agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt     3540

taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt atgtaaacaa     3600

tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc     3660

ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt taattaaata     3720

caaaggatac caggtggccc ccgctgaatt ggagtcgata ttgttacaac accccaacat     3780

cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt     3840

tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag     3900

tcaagtaaca accgccaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa     3960

aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa     4020

gggcggaaag tccaaattgt aaataacttc gtatagcata cattatacga agttatatga     4080

tatcggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt     4140

ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga     4200

aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag     4260

tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagggtac     4320

cgccaccatg gggtaccgcc accatgggat cggccattga acaagatgga ttgcacgcag     4380

gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg     4440

gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca     4500

agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc     4560

tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg     4620

actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg     4680

ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta     4740

cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag     4800

ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac     4860

tgttcgccag gctcaaggcg cgcatgcccg acggcgatga tctcgtcgtg acccatggcg     4920

atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg     4980

gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg     5040

aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg     5100

attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagtc gacacgcgtt     5160

ggtaccataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga     5220

tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg     5280

ctgtccccag tgcaagtgca ggtgccagaa catttctcta taacttcgta tagcatacat     5340

tatacgaagt tatgacagat tgcctttcct ttaggcaggt aatcagggcc tgagtgcttt     5400

acctttaaaa tcattccctt tcccatccat aaggtgcagt aggagcaaga gagggcaagt     5460

gttcccaaat tgacgccacc ataataatca tcaccacacc gtgagcagat ctgaaaggca     5520

cactttgatt tttttaagga taagaaccac agaacactgg gtagtagcta atgaaattga     5580

gaagggaaat cttagcatgc ctccaaaaat tcacatccaa tgcatagttt gttcaaattt     5640

aaggttactc aggcattgat cttttcagtg ttttttcact ttagctatgt ggattagcta     5700

gaatgcacac caaaaaaata cttgagctgt atatatatat gtgtgtgtgt gtgtgtgtgt     5760

gtgtgtgtgt gtgtgcatgt atgtgcacat gtgtctgtgt ggtatatttg tgtatgtgta     5820

tttgtatgta ctgttattga aaatatattt aatacctttg gaaaaatctt gggcaagatg     5880

acctactagt tttccttgaa aaaaagttgc tttgttatta atattgtgct taaattattt     5940

ttatacacca ttgttcctta cctttacata attgcaatat ttccccctta ctacttcttg     6000

gaaaaaaatt acaaaatgaa gttttataga aaagatggat ttgctttgct tggtttttct     6060

tatttgagct accaagagga gagaactctg attaaatatc ttgtgattta aatcctgccc     6120

cctccccaag aaggtgggga cgttcagaat cacacttgaa gagggggtgc attgcctggt     6180

cccagggaga aggagggagc agatggggag gaatgaggca gggaaccatg gttggggttt     6240

tctgaatttg ctgaagccac tagttggcat cctttaacaa ttctacaaac ctttagatgg     6300

tgaggcttat gtggcaggac tgggagggtt taatgcatgc tttttgacag tttggatctg     6360

gaggggtctc aggcttccac aaaaattaca ttgtggacag tttcatagga ttttgccttt     6420

taactcttca aagcaacagt tttctcaact gtcatccccg cgggggagca gaggaagaca     6480

agtgagaagg aaaaggggac aggatctggg ttgagcaagc attaattgag cgccttctgt     6540

atcccggaca tcatgccaac accatgggct gcccgagaag agaggtgggt ggtttggtcc     6600

ccagctctgc cttgaaccct cacctgctct cctctactcc cagtgtttat tggatattcc     6660

agaacagtcc aaacgaacca taaactttat atcacagatt caatctgtgt aaatactatt     6720

atcttcaagt tgtagtttct ttcttgggcc caggctgtga aattggcttc acccctacca     6780

caccccagcg ggcaggcttg ttcttatgtc acactccacc cctccagtct cctcaactgg     6840

aaaacttgcc tcttttcctc acttcctctg ctccttacaa tggcttcata tggaaggtga     6900

agctgagatc accccaattt tacaggtgag gaaactgagg tcaagctagg agcgtccagc     6960

ggggctgaga aattgccagg atttccttcc ccattaagga gctgagagtc ttagctccag     7020

acac                                                                  7024


<210>  5
<211>  825
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  P2A-mScarlet-T2A-Luciferase

<400>  5

Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val 
1               5                   10                  15      


Glu Glu Asn Pro Gly Pro Met Val Ser Lys Gly Glu Ala Val Ile Lys 
            20                  25                  30          


Glu Phe Met Arg Phe Lys Val His Met Glu Gly Ser Met Asn Gly His 
        35                  40                  45              


Glu Phe Glu Ile Glu Gly Glu Gly Glu Gly Arg Pro Tyr Glu Gly Thr 
    50                  55                  60                  


Gln Thr Ala Lys Leu Lys Val Thr Lys Gly Gly Pro Leu Pro Phe Ser 
65                  70                  75                  80  


Trp Asp Ile Leu Ser Pro Gln Phe Met Tyr Gly Ser Arg Ala Phe Ile 
                85                  90                  95      


Lys His Pro Ala Asp Ile Pro Asp Tyr Tyr Lys Gln Ser Phe Pro Glu 
            100                 105                 110         


Gly Phe Lys Trp Glu Arg Val Met Asn Phe Glu Asp Gly Gly Ala Val 
        115                 120                 125             


Thr Val Thr Gln Asp Thr Ser Leu Glu Asp Gly Thr Leu Ile Tyr Lys 
    130                 135                 140                 


Val Lys Leu Arg Gly Thr Asn Phe Pro Pro Asp Gly Pro Val Met Gln 
145                 150                 155                 160 


Lys Lys Thr Met Gly Trp Glu Ala Ser Thr Glu Arg Leu Tyr Pro Glu 
                165                 170                 175     


Asp Gly Val Leu Lys Gly Asp Ile Lys Met Ala Leu Arg Leu Lys Asp 
            180                 185                 190         


Gly Gly Arg Tyr Leu Ala Asp Phe Lys Thr Thr Tyr Lys Ala Lys Lys 
        195                 200                 205             


Pro Val Gln Met Pro Gly Ala Tyr Asn Val Asp Arg Lys Leu Asp Ile 
    210                 215                 220                 


Thr Ser His Asn Glu Asp Tyr Thr Val Val Glu Gln Tyr Glu Arg Ser 
225                 230                 235                 240 


Glu Gly Arg His Ser Thr Gly Gly Met Asp Glu Leu Tyr Lys Gly Ser 
                245                 250                 255     


Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn 
            260                 265                 270         


Pro Gly Pro Met Glu Asp Ala Lys Asn Ile Lys Lys Gly Pro Ala Pro 
        275                 280                 285             


Phe Tyr Pro Leu Glu Asp Gly Thr Ala Gly Glu Gln Leu His Lys Ala 
    290                 295                 300                 


Met Lys Arg Tyr Ala Leu Val Pro Gly Thr Ile Ala Phe Thr Asp Ala 
305                 310                 315                 320 


His Ile Glu Val Asn Ile Thr Tyr Ala Glu Tyr Phe Glu Met Ser Val 
                325                 330                 335     


Arg Leu Ala Glu Ala Met Lys Arg Tyr Gly Leu Asn Thr Asn His Arg 
            340                 345                 350         


Ile Val Val Cys Ser Glu Asn Ser Leu Gln Phe Phe Met Pro Val Leu 
        355                 360                 365             


Gly Ala Leu Phe Ile Gly Val Ala Val Ala Pro Ala Asn Asp Ile Tyr 
    370                 375                 380                 


Asn Glu Arg Glu Leu Leu Asn Ser Met Asn Ile Ser Gln Pro Thr Val 
385                 390                 395                 400 


Val Phe Val Ser Lys Lys Gly Leu Gln Lys Ile Leu Asn Val Gln Lys 
                405                 410                 415     


Lys Leu Pro Ile Ile Gln Lys Ile Ile Ile Met Asp Ser Lys Thr Asp 
            420                 425                 430         


Tyr Gln Gly Phe Gln Ser Met Tyr Thr Phe Val Thr Ser His Leu Pro 
        435                 440                 445             


Pro Gly Phe Asn Glu Tyr Asp Phe Val Pro Glu Ser Phe Asp Arg Asp 
    450                 455                 460                 


Lys Thr Ile Ala Leu Ile Met Asn Ser Ser Gly Ser Thr Gly Leu Pro 
465                 470                 475                 480 


Lys Gly Val Ala Leu Pro His Arg Thr Ala Cys Val Arg Phe Ser His 
                485                 490                 495     


Ala Arg Asp Pro Ile Phe Gly Asn Gln Ile Ile Pro Asp Thr Ala Ile 
            500                 505                 510         


Leu Ser Val Val Pro Phe His His Gly Phe Gly Met Phe Thr Thr Leu 
        515                 520                 525             


Gly Tyr Leu Ile Cys Gly Phe Arg Val Val Leu Met Tyr Arg Phe Glu 
    530                 535                 540                 


Glu Glu Leu Phe Leu Arg Ser Leu Gln Asp Tyr Lys Ile Gln Ser Ala 
545                 550                 555                 560 


Leu Leu Val Pro Thr Leu Phe Ser Phe Phe Ala Lys Ser Thr Leu Ile 
                565                 570                 575     


Asp Lys Tyr Asp Leu Ser Asn Leu His Glu Ile Ala Ser Gly Gly Ala 
            580                 585                 590         


Pro Leu Ser Lys Glu Val Gly Glu Ala Val Ala Lys Arg Phe His Leu 
        595                 600                 605             


Pro Gly Ile Arg Gln Gly Tyr Gly Leu Thr Glu Thr Thr Ser Ala Ile 
    610                 615                 620                 


Leu Ile Thr Pro Glu Gly Asp Asp Lys Pro Gly Ala Val Gly Lys Val 
625                 630                 635                 640 


Val Pro Phe Phe Glu Ala Lys Val Val Asp Leu Asp Thr Gly Lys Thr 
                645                 650                 655     


Leu Gly Val Asn Gln Arg Gly Glu Leu Cys Val Arg Gly Pro Met Ile 
            660                 665                 670         


Met Ser Gly Tyr Val Asn Asn Pro Glu Ala Thr Asn Ala Leu Ile Asp 
        675                 680                 685             


Lys Asp Gly Trp Leu His Ser Gly Asp Ile Ala Tyr Trp Asp Glu Asp 
    690                 695                 700                 


Glu His Phe Phe Ile Val Asp Arg Leu Lys Ser Leu Ile Lys Tyr Lys 
705                 710                 715                 720 


Gly Tyr Gln Val Ala Pro Ala Glu Leu Glu Ser Ile Leu Leu Gln His 
                725                 730                 735     


Pro Asn Ile Phe Asp Ala Gly Val Ala Gly Leu Pro Asp Asp Asp Ala 
            740                 745                 750         


Gly Glu Leu Pro Ala Ala Val Val Val Leu Glu His Gly Lys Thr Met 
        755                 760                 765             


Thr Glu Lys Glu Ile Val Asp Tyr Val Ala Ser Gln Val Thr Thr Ala 
    770                 775                 780                 


Lys Lys Leu Arg Gly Gly Val Val Phe Val Asp Glu Val Pro Lys Gly 
785                 790                 795                 800 


Leu Thr Gly Lys Leu Asp Ala Arg Lys Ile Arg Glu Ile Leu Ile Lys 
                805                 810                 815     


Ala Lys Lys Gly Gly Lys Ser Lys Leu 
            820                 825 


<210>  6
<211>  1561
<212>  DNA
<213>  Homo sapiens

<400>  6
ccaatgccat caagaccatc cagaagaaca caatctttac caatgtggcc gagaccagcg       60

acgggggcgt ttactgggaa ggcattgatg agccgctagc ttcaggtgtc accatcacgt      120

cctggaagaa taaggagtgg agctcagagg atggtgtgtc cctgccagag gccttggtgt      180

gccgggctgc agggactgcc tgtttgagcc aggcactcac gagcctttct ctgtcttata      240

ggggaacctt gtgcccaccc caactcgagg ttctgcaccc ctgccagcca gtgccccatc      300

attgatgctg cctgggagtc tccggaaggt gttcccattg aaggcattat ctttggaggc      360

cgtagacctg ctggtgaggc tctccttcat ttaggctggg aacatgggtg tgctgggtac      420

cgaagggcat ctgtgaaatc tctccttttc catgaccttg tcagagggtg cccaggggct      480

tcctttcttg agcttccttc ccaaagatcc agaataattg gcaagttcaa atgtagaacc      540

aacccttctg gtcacctgga acctttctga atccctgatc tattgtagct tgatcaaatt      600

ttacttttta ctttgtggcc tcagtcatgt aactttgagt tagcagtttt ctgcaattta      660

gcttggtgaa cgcaaaacta gctcgattac aaagttattg tcttgccgtg tctttccgtg      720

ttgtgaataa caccactggt tgtggagtct gaatttcaaa gcctctgatg aacatttctc      780

tttttttttc ctgctaaagg tgtccctcta gtctatgaag ctctcagctg gcaacatgga      840

gtctttgtgg gggcggccat gagatcagag gccacagcgg ctgcagaaca taaaggtaaa      900

tcaaagtcct gatctgaaac cacagagaag tgggattaga gcactcttcg tcactcttat      960

gtctctctcc ttttctgtgt ctgtgtgtgg ggagagagag agagagaaag agagagagga     1020

gaacaaagca tgctaatgtc aacaatcaat ggcgtcagtc ttgcctagga gagcctcatt     1080

tactaatgaa ctccctctct gtttaacagg caaaatcatc atgcatgacc cctttgccat     1140

gcggcccttc tttggctaca acttcggcaa atacctggcc cactggctta gcatggccca     1200

gcacccagca gccaaactgc ccaagatctt ccatgtcaac tggttccgga aggacaagga     1260

aggcaaattc ctctggccag gctttggaga gaactccagg gtgctggagt ggatgttcaa     1320

ccggatcgat ggaaaagcca gcaccaagct cacgcccata ggctacatcc ccaaggagga     1380

tgccctgaac ctgaaaggcc tggggcacat caacatgatg gagcttttca gcatctccaa     1440

ggaattctgg gagaaggagg tggaagacat cgagaagtat ctggaggatc aagtcaatgc     1500

cgacctcccc tgtgaaatcg agagagagat ccttgccttg aagcaaagaa taagccagat     1560

g                                                                     1561


<210>  7
<211>  66
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  P2A nucleic acid

<400>  7
ggatccggag ctactaactt cagcctgctg aagcaggctg gagatgtgga ggagaaccct       60

ggacct                                                                  66


<210>  8
<211>  696
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mScarlet nucleic acid

<400>  8
atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag       60

ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag      120

ggcacccaga ccgccaagct gaaggtgaca aagggtggcc ccctgccctt ctcctgggac      180

atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc      240

cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc      300

gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc      360

tacaaggtga agctgcgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag      420

acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc      480

gacattaaga tggccctgcg cctgaaggac ggcgggcgct acctggcgga cttcaagacc      540

acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtgga ccgcaagttg      600

gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc      660

cgccactcca ccggcggcat ggacgagctg tacaag                                696


<210>  9
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  T2A nucleic acid

<400>  9
ggcagcggag agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc       60

cct                                                                     63


<210>  10
<211>  1653
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Luciferase nucleic acid

<400>  10
atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct agaggatgga       60

accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaaccatt      120

gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc      180

gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta      240

tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt      300

gcagttgcgc ccgcgaacga catctataat gaacgtgaat tgctcaacag tatgaacatt      360

tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa      420

aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga      480

tttcagtcga tgtacacctt cgtcacatct catctacctc ccggttttaa tgaatacgat      540

tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa ctcctctgga      600

tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg      660

catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt      720

gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt      780

cgagtcgtct taatgtatag atttgaagag gagctgtttt tacgatccct tcaggattac      840

aaaatccaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg      900

attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg      960

aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat     1020

gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc     1080

gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa     1140

acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt     1200

tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct     1260

ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg cttgaagtct     1320

ttaattaaat acaaaggata ccaggtggcc cccgctgaat tggagtcgat attgttacaa     1380

caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc cggtgaactt     1440

cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat     1500

tacgtcgcca gtcaagtaac aaccgccaaa aagttgcgcg gaggagttgt gtttgtggac     1560

gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata     1620

aaggccaaga agggcggaaa gtccaaattg taa                                  1653


<210>  11
<211>  1243
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  NeoR cassette

<400>  11
atgatatcgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga       60

agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact      120

gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata      180

taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg      240

gtaccgccac catggggtac cgccaccatg ggatcggcca ttgaacaaga tggattgcac      300

gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca      360

atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt      420

gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg      480

tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga      540

agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct      600

cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg      660

gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg      720

gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc      780

gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg atgatctcgt cgtgacccat      840

ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac      900

tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt      960

gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct     1020

cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agtcgacacg     1080

cgttggtacc ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa     1140

tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa     1200

taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tct                       1243


<210>  12
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LoxP nucleic acid

<400>  12
ataacttcgt atagcataca ttatacgaag ttat                                   34


<210>  13
<211>  1646
<212>  DNA
<213>  Homo sapiens

<400>  13
gtaatcaggg cctgagtgct ttacctttaa aatcattccc tttcccatcc ataaggtgca       60

gtaggagcaa gagagggcaa gtgttcccaa attgacgcca ccataataat catcaccaca      120

ccgtgagcag atctgaaagg cacactttga tttttttaag gataagaacc acagaacact      180

gggtagtagc taatgaaatt gagaagggaa atcttagcat gcctccaaaa attcacatcc      240

aatgcatagt ttgttcaaat ttaaggttac tcaggcattg atcttttcag tgttttttca      300

ctttagctat gtggattagc tagaatgcac accaaaaaaa tacttgagct gtatatatat      360

atgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgcat gtatgtgcac atgtgtctgt      420

gtggtatatt tgtgtatgtg tatttgtatg tactgttatt gaaaatatat ttaatacctt      480

tggaaaaatc ttgggcaaga tgacctacta gttttccttg aaaaaaagtt gctttgttat      540

taatattgtg cttaaattat ttttatacac cattgttcct tacctttaca taattgcaat      600

atttccccct tactacttct tggaaaaaaa ttacaaaatg aagttttata gaaaagatgg      660

atttgctttg cttggttttt cttatttgag ctaccaagag gagagaactc tgattaaata      720

tcttgtgatt taaatcctgc cccctcccca agaaggtggg gacgttcaga atcacacttg      780

aagagggggt gcattgcctg gtcccaggga gaaggaggga gcagatgggg aggaatgagg      840

cagggaacca tggttggggt tttctgaatt tgctgaagcc actagttggc atcctttaac      900

aattctacaa acctttagat ggtgaggctt atgtggcagg actgggaggg tttaatgcat      960

gctttttgac agtttggatc tggaggggtc tcaggcttcc acaaaaatta cattgtggac     1020

agtttcatag gattttgcct tttaactctt caaagcaaca gttttctcaa ctgtcatccc     1080

cgcgggggag cagaggaaga caagtgagaa ggaaaagggg acaggatctg ggttgagcaa     1140

gcattaattg agcgccttct gtatcccgga catcatgcca acaccatggg ctgcccgaga     1200

agagaggtgg gtggtttggt ccccagctct gccttgaacc ctcacctgct ctcctctact     1260

cccagtgttt attggatatt ccagaacagt ccaaacgaac cataaacttt atatcacaga     1320

ttcaatctgt gtaaatacta ttatcttcaa gttgtagttt ctttcttggg cccaggctgt     1380

gaaattggct tcacccctac cacaccccag cgggcaggct tgttcttatg tcacactcca     1440

cccctccagt ctcctcaact ggaaaacttg cctcttttcc tcacttcctc tgctccttac     1500

aatggcttca tatggaaggt gaagctgaga tcaccccaat tttacaggtg aggaaactga     1560

ggtcaagcta ggagcgtcca gcggggctga gaaattgcca ggatttcctt ccccattaag     1620

gagctgagag tcttagctcc agacac                                          1646


<210>  14
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  P2A peptide

<400>  14

Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val 
1               5                   10                  15      


Glu Glu Asn Pro Gly Pro 
            20          


<210>  15
<211>  232
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  mScarlet peptide

<400>  15

Met Val Ser Lys Gly Glu Ala Val Ile Lys Glu Phe Met Arg Phe Lys 
1               5                   10                  15      


Val His Met Glu Gly Ser Met Asn Gly His Glu Phe Glu Ile Glu Gly 
            20                  25                  30          


Glu Gly Glu Gly Arg Pro Tyr Glu Gly Thr Gln Thr Ala Lys Leu Lys 
        35                  40                  45              


Val Thr Lys Gly Gly Pro Leu Pro Phe Ser Trp Asp Ile Leu Ser Pro 
    50                  55                  60                  


Gln Phe Met Tyr Gly Ser Arg Ala Phe Ile Lys His Pro Ala Asp Ile 
65                  70                  75                  80  


Pro Asp Tyr Tyr Lys Gln Ser Phe Pro Glu Gly Phe Lys Trp Glu Arg 
                85                  90                  95      


Val Met Asn Phe Glu Asp Gly Gly Ala Val Thr Val Thr Gln Asp Thr 
            100                 105                 110         


Ser Leu Glu Asp Gly Thr Leu Ile Tyr Lys Val Lys Leu Arg Gly Thr 
        115                 120                 125             


Asn Phe Pro Pro Asp Gly Pro Val Met Gln Lys Lys Thr Met Gly Trp 
    130                 135                 140                 


Glu Ala Ser Thr Glu Arg Leu Tyr Pro Glu Asp Gly Val Leu Lys Gly 
145                 150                 155                 160 


Asp Ile Lys Met Ala Leu Arg Leu Lys Asp Gly Gly Arg Tyr Leu Ala 
                165                 170                 175     


Asp Phe Lys Thr Thr Tyr Lys Ala Lys Lys Pro Val Gln Met Pro Gly 
            180                 185                 190         


Ala Tyr Asn Val Asp Arg Lys Leu Asp Ile Thr Ser His Asn Glu Asp 
        195                 200                 205             


Tyr Thr Val Val Glu Gln Tyr Glu Arg Ser Glu Gly Arg His Ser Thr 
    210                 215                 220                 


Gly Gly Met Asp Glu Leu Tyr Lys 
225                 230         


<210>  16
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  T2A peptide

<400>  16

Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu 
1               5                   10                  15      


Glu Asn Pro Gly Pro 
            20      


<210>  17
<211>  550
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Luciferase peptide

<400>  17

Met Glu Asp Ala Lys Asn Ile Lys Lys Gly Pro Ala Pro Phe Tyr Pro 
1               5                   10                  15      


Leu Glu Asp Gly Thr Ala Gly Glu Gln Leu His Lys Ala Met Lys Arg 
            20                  25                  30          


Tyr Ala Leu Val Pro Gly Thr Ile Ala Phe Thr Asp Ala His Ile Glu 
        35                  40                  45              


Val Asn Ile Thr Tyr Ala Glu Tyr Phe Glu Met Ser Val Arg Leu Ala 
    50                  55                  60                  


Glu Ala Met Lys Arg Tyr Gly Leu Asn Thr Asn His Arg Ile Val Val 
65                  70                  75                  80  


Cys Ser Glu Asn Ser Leu Gln Phe Phe Met Pro Val Leu Gly Ala Leu 
                85                  90                  95      


Phe Ile Gly Val Ala Val Ala Pro Ala Asn Asp Ile Tyr Asn Glu Arg 
            100                 105                 110         


Glu Leu Leu Asn Ser Met Asn Ile Ser Gln Pro Thr Val Val Phe Val 
        115                 120                 125             


Ser Lys Lys Gly Leu Gln Lys Ile Leu Asn Val Gln Lys Lys Leu Pro 
    130                 135                 140                 


Ile Ile Gln Lys Ile Ile Ile Met Asp Ser Lys Thr Asp Tyr Gln Gly 
145                 150                 155                 160 


Phe Gln Ser Met Tyr Thr Phe Val Thr Ser His Leu Pro Pro Gly Phe 
                165                 170                 175     


Asn Glu Tyr Asp Phe Val Pro Glu Ser Phe Asp Arg Asp Lys Thr Ile 
            180                 185                 190         


Ala Leu Ile Met Asn Ser Ser Gly Ser Thr Gly Leu Pro Lys Gly Val 
        195                 200                 205             


Ala Leu Pro His Arg Thr Ala Cys Val Arg Phe Ser His Ala Arg Asp 
    210                 215                 220                 


Pro Ile Phe Gly Asn Gln Ile Ile Pro Asp Thr Ala Ile Leu Ser Val 
225                 230                 235                 240 


Val Pro Phe His His Gly Phe Gly Met Phe Thr Thr Leu Gly Tyr Leu 
                245                 250                 255     


Ile Cys Gly Phe Arg Val Val Leu Met Tyr Arg Phe Glu Glu Glu Leu 
            260                 265                 270         


Phe Leu Arg Ser Leu Gln Asp Tyr Lys Ile Gln Ser Ala Leu Leu Val 
        275                 280                 285             


Pro Thr Leu Phe Ser Phe Phe Ala Lys Ser Thr Leu Ile Asp Lys Tyr 
    290                 295                 300                 


Asp Leu Ser Asn Leu His Glu Ile Ala Ser Gly Gly Ala Pro Leu Ser 
305                 310                 315                 320 


Lys Glu Val Gly Glu Ala Val Ala Lys Arg Phe His Leu Pro Gly Ile 
                325                 330                 335     


Arg Gln Gly Tyr Gly Leu Thr Glu Thr Thr Ser Ala Ile Leu Ile Thr 
            340                 345                 350         


Pro Glu Gly Asp Asp Lys Pro Gly Ala Val Gly Lys Val Val Pro Phe 
        355                 360                 365             


Phe Glu Ala Lys Val Val Asp Leu Asp Thr Gly Lys Thr Leu Gly Val 
    370                 375                 380                 


Asn Gln Arg Gly Glu Leu Cys Val Arg Gly Pro Met Ile Met Ser Gly 
385                 390                 395                 400 


Tyr Val Asn Asn Pro Glu Ala Thr Asn Ala Leu Ile Asp Lys Asp Gly 
                405                 410                 415     


Trp Leu His Ser Gly Asp Ile Ala Tyr Trp Asp Glu Asp Glu His Phe 
            420                 425                 430         


Phe Ile Val Asp Arg Leu Lys Ser Leu Ile Lys Tyr Lys Gly Tyr Gln 
        435                 440                 445             


Val Ala Pro Ala Glu Leu Glu Ser Ile Leu Leu Gln His Pro Asn Ile 
    450                 455                 460                 


Phe Asp Ala Gly Val Ala Gly Leu Pro Asp Asp Asp Ala Gly Glu Leu 
465                 470                 475                 480 


Pro Ala Ala Val Val Val Leu Glu His Gly Lys Thr Met Thr Glu Lys 
                485                 490                 495     


Glu Ile Val Asp Tyr Val Ala Ser Gln Val Thr Thr Ala Lys Lys Leu 
            500                 505                 510         


Arg Gly Gly Val Val Phe Val Asp Glu Val Pro Lys Gly Leu Thr Gly 
        515                 520                 525             


Lys Leu Asp Ala Arg Lys Ile Arg Glu Ile Leu Ile Lys Ala Lys Lys 
    530                 535                 540                 


Gly Gly Lys Ser Lys Leu 
545                 550 


<210>  18
<211>  267
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  NeoR peptide

<400>  18

Met Gly Ser Ala Ile Glu Gln Asp Gly Leu His Ala Gly Ser Pro Ala 
1               5                   10                  15      


Ala Trp Val Glu Arg Leu Phe Gly Tyr Asp Trp Ala Gln Gln Thr Ile 
            20                  25                  30          


Gly Cys Ser Asp Ala Ala Val Phe Arg Leu Ser Ala Gln Gly Arg Pro 
        35                  40                  45              


Val Leu Phe Val Lys Thr Asp Leu Ser Gly Ala Leu Asn Glu Leu Gln 
    50                  55                  60                  


Asp Glu Ala Ala Arg Leu Ser Trp Leu Ala Thr Thr Gly Val Pro Cys 
65                  70                  75                  80  


Ala Ala Val Leu Asp Val Val Thr Glu Ala Gly Arg Asp Trp Leu Leu 
                85                  90                  95      


Leu Gly Glu Val Pro Gly Gln Asp Leu Leu Ser Ser His Leu Ala Pro 
            100                 105                 110         


Ala Glu Lys Val Ser Ile Met Ala Asp Ala Met Arg Arg Leu His Thr 
        115                 120                 125             


Leu Asp Pro Ala Thr Cys Pro Phe Asp His Gln Ala Lys His Arg Ile 
    130                 135                 140                 


Glu Arg Ala Arg Thr Arg Met Glu Ala Gly Leu Val Asp Gln Asp Asp 
145                 150                 155                 160 


Leu Asp Glu Glu His Gln Gly Leu Ala Pro Ala Glu Leu Phe Ala Arg 
                165                 170                 175     


Leu Lys Ala Arg Met Pro Asp Gly Asp Asp Leu Val Val Thr His Gly 
            180                 185                 190         


Asp Ala Cys Leu Pro Asn Ile Met Val Glu Asn Gly Arg Phe Ser Gly 
        195                 200                 205             


Phe Ile Asp Cys Gly Arg Leu Gly Val Ala Asp Arg Tyr Gln Asp Ile 
    210                 215                 220                 


Ala Leu Ala Thr Arg Asp Ile Ala Glu Glu Leu Gly Gly Glu Trp Ala 
225                 230                 235                 240 


Asp Arg Phe Leu Val Leu Tyr Gly Ile Ala Ala Pro Asp Ser Gln Arg 
                245                 250                 255     


Ile Ala Phe Tyr Arg Leu Leu Asp Glu Phe Phe 
            260                 265         


