                         SEQUENCE LISTING

<110>  UCL BUSINESS PLC.
 
<120>  PRODUCTION OF THERAPEUTIC CELLS

<130>  N409552WO

<140>  PCT/GB2017/053862
<141>  2017-12-21

<150>  GB 1621874.5
<151>  2016-12-21

<150>  GB 1706101.1
<151>  2017-04-18

<160>  19    

<170>  PatentIn version 3.5

<210>  1
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA for TRAC

<400>  1
tctctcagct ggtacacggc                                                   20


<210>  2
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TRAC forward primer

<400>  2
ttgatagctt gtgcctgtcc c                                                 21


<210>  3
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TRAC reverse primer

<400>  3
ggcaaacagt ctgagcaaag g                                                 21


<210>  4
<211>  600
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Terminal CRISPR long terminal repeat

<400>  4
agagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga       60

gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt      120

agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat      180

catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa      240

ggacgaaaca ccgggtcttc gatggaagac ctgttttaga gctagaaata gcaagttaaa      300

ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctaga      360

ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct      420

ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta      480

gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca      540

gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt      600


<210>  5
<211>  187
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  U6 promoter region sequenced in Example 1

<400>  5
aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa       60

atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta      120

aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata      180

tcttgtg                                                                187


<210>  6
<211>  187
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  U6 promoter region sequenced in Example 1


<220>
<221>  misc_feature
<222>  (73)..(73)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (131)..(131)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (133)..(133)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (162)..(162)
<223>  n = unknown nucleotide

<400>  6
aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa       60

atacgtgacg tanaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta      120

aaatggacta ncntatgctt accgtaactt gaaagtattt cnatttcttg gctttatata      180

tcttgtg                                                                187


<210>  7
<211>  62
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  U6 promotor sequence determined in Example 1


<220>
<221>  MISC_FEATURE
<222>  (3)..(3)
<223>  x = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (7)..(7)
<223>  x = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (9)..(9)
<223>  x = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  x = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (40)..(40)
<223>  x = unknown amino acid

<400>  7

Gly Cys Xaa Arg Asp Asn Xaa Asn Xaa Phe Asp Cys Lys His Lys Asp 
1               5                   10                  15      


Ile Ser Thr Lys Tyr Val Thr Xaa Lys Val Ile Ile Ser Trp Val Val 
            20                  25                  30          


Cys Ser Phe Lys Ile Met Phe Xaa Asn Gly Leu Ser Tyr Ala Tyr Arg 
        35                  40                  45              


Asn Leu Lys Val Phe Arg Phe Leu Gly Phe Ile Tyr Leu Val 
    50                  55                  60          


<210>  8
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  U6 promoter sequence determined in in Example 1


<220>
<221>  misc_feature
<222>  (7)..(7)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (65)..(65)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (67)..(67)
<223>  n = unknown nucleotide

<400>  8
gacgtanaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg       60

actancntat                                                              70


<210>  9
<211>  245
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  misc_feature
<222>  (1)..(29)
<223>  gRNA scaffold

<220>
<221>  misc_feature
<222>  (30)..(50)
<223>  HIV-deltaU3

<220>
<221>  misc_feature
<222>  (51)..(146)
<223>  HIV- R region

<220>
<221>  misc_feature
<222>  (147)..(228)
<223>  HIV- U5

<220>
<221>  misc_feature
<222>  (232)..(245)
<223>  HIV - PBS

<400>  9
tggcaccgag tcggtgcttt ttttctagac tgctttttgc ttgtactggg tctctctggt       60

tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc      120

aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta      180

actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa      240

caggg                                                                  245


<210>  10
<211>  316
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  misc_feature
<222>  (1)..(29)
<223>  gRNA scaffold

<220>
<221>  misc_feature
<222>  (30)..(50)
<223>  HIV-deltaU3

<220>
<221>  misc_feature
<222>  (51)..(146)
<223>  HIV - R region

<220>
<221>  misc_feature
<222>  (147)..(228)
<223>  HIV-U5

<220>
<221>  misc_feature
<222>  (232)..(249)
<223>  HIV - PBS

<220>
<221>  misc_feature
<222>  (282)..(316)
<223>  HIV - Psi

<400>  10
tggcaccgag tcggtgcttt ttttctagac tgctttttgc ttgtactggg tctctctggt       60

tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc      120

aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta      180

actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa      240

cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc      300

tgaagcgcgc acggca                                                      316


<210>  11
<211>  286
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  HIV-deltaU3

<220>
<221>  misc_feature
<222>  (21)..(116)
<223>  HIV - R region

<220>
<221>  misc_feature
<222>  (117)..(190)
<223>  HIV - U5

<220>
<221>  misc_feature
<222>  (202)..(219)
<223>  HIV - PBS

<220>
<221>  misc_feature
<222>  (252)..(286)
<223>  HIV - Psi

<400>  11
tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg       60

gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag      120

tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag      180

tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga      240

ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggca                     286


<210>  12
<211>  105
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  MISC_FEATURE
<222>  (9)..(9)
<223>  Xaa =  unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (38)..(38)
<223>  Xaa =  unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (57)..(57)
<223>  Xaa =  unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (60)..(60)
<223>  Xaa =  unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (75)..(75)
<223>  Xaa =  unknown amino acid

<400>  12

Gly Thr Glu Ser Val Leu Phe Phe Xaa Thr Ala Phe Cys Leu Tyr Trp 
1               5                   10                  15      


Val Ser Leu Val Arg Pro Asp Leu Ser Leu Gly Ala Leu Trp Leu Thr 
            20                  25                  30          


Arg Glu Pro Thr Ala Xaa Ala Ser Ile Lys Leu Ala Leu Ser Ala Ser 
        35                  40                  45              


Ser Ser Val Cys Pro Ser Val Val Xaa Leu Trp Xaa Leu Glu Ile Pro 
    50                  55                  60                  


Gln Thr Leu Leu Val Ser Val Glu Asn Leu Xaa Gln Trp Arg Pro Asn 
65                  70                  75                  80  


Arg Asp Leu Lys Ala Lys Gly Lys Pro Glu Glu Leu Ser Arg Arg Arg 
                85                  90                  95      


Thr Arg Leu Ala Glu Ala Arg Thr Ala 
            100                 105 


<210>  13
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1

<400>  13
agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa       60

gcttgccttg                                                              70


<210>  14
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1

<400>  14
agaggagctc tctcgacgca ggactcgggt tgctgaagcg ggcacggcaa gaggcgaggg       60

gcggcgactg                                                              70


<210>  15
<211>  332
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  misc_feature
<222>  (1)..(47)
<223>  HIV - deltaU3

<220>
<221>  misc_feature
<222>  (48)..(143)
<223>  HIV - R region

<220>
<221>  misc_feature
<222>  (144)..(225)
<223>  HIV - U5

<220>
<221>  misc_feature
<222>  (229)..(246)
<223>  HIV - PBS

<220>
<221>  misc_feature
<222>  (279)..(332)
<223>  HIV - Psi

<400>  15
taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag       60

accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat      120

aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact      180

agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag      240

ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga      300

agcgcgcacg gcggggcggc gacaagaggc ga                                    332


<210>  16
<211>  332
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  misc_feature
<222>  (1)..(47)
<223>  HIV - deltaU3

<220>
<221>  misc_feature
<222>  (3)..(5)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (7)..(8)
<223>  n = unknown nucleotide

<220>
<221>  misc_feature
<222>  (48)..(143)
<223>  HIV - R region

<220>
<221>  misc_feature
<222>  (144)..(225)
<223>  HIV - U5

<220>
<221>  misc_feature
<222>  (229)..(246)
<223>  HIV - PBS

<220>
<221>  misc_feature
<222>  (279)..(332)
<223>  HIV - Psi

<400>  16
tannncnntc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag       60

accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat      120

aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact      180

agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag      240

ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga      300

agcgcgcacg gcggggcggc gacaagaggc ga                                    332


<210>  17
<211>  112
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sequence determined in Example 1


<220>
<221>  MISC_FEATURE
<222>  (37)..(37)
<223>  Xaa = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (57)..(57)
<223>  Xaa = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (60)..(60)
<223>  Xaa = unknown amino acid

<220>
<221>  MISC_FEATURE
<222>  (75)..(75)
<223>  Xaa = unknown amino acid

<400>  17

Asn Ser Leu Pro Thr Lys Thr Arg Ser Ala Phe Cys Leu Tyr Trp Val 
1               5                   10                  15      


Ser Leu Val Arg Pro Asp Leu Ser Leu Gly Ala Leu Trp Leu Thr Arg 
            20                  25                  30          


Glu Pro Thr Ala Xaa Ala Ser Ile Lys Leu Ala Leu Ser Ala Ser Ser 
        35                  40                  45              


Ser Val Cys Pro Pro Ser Val Val Xaa Leu Trp Xaa Leu Glu Ile Pro 
    50                  55                  60                  


Gln Thr Leu Leu Val Ser Val Glu Asn Leu Xaa Gln Trp Arg Pro Asn 
65                  70                  75                  80  


Arg Asp Leu Lys Ala Lys Gly Lys Pro Glu Glu Leu Ser Arg Arg Arg 
                85                  90                  95      


Thr Arg Leu Ala Glu Ala Arg Thr Ala Arg Gly Glu Gly Arg Arg Leu 
            100                 105                 110         


<210>  18
<211>  630
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (174)..(180)
<223>  Splice acceptor site

<220>
<221>  misc_feature
<222>  (181)..(453)
<223>  EX1

<220>
<221>  misc_feature
<222>  (435)..(438)
<223>  PAM

<220>
<221>  misc_feature
<222>  (439)..(458)
<223>  Cas9-CD donor site guide

<220>
<221>  misc_feature
<222>  (453)..(459)
<223>  Splice donor site

<400>  18
cccagtccat cacgagcagc tggtttctaa gatgctattt cccgtataaa gcatgagacc       60

gtgacttgcc agccccacag agccccgccc ttgtccatca ctggcatctg gactccagcc      120

tgggttgggg caaagaggga aatgagatca tgtcctaacc ctgatcctct tgtcccacag      180

atatccagaa ccctgaccct gccgtgtacc agctgagaga ctctaaatcc agtgacaagt      240

ctgtctgcct attcaccgat tttgattctc aaacaaatgt gtcacaaagt aaggattctg      300

atgtgtatat cacagacaaa actgtgctag acatgaggtc tatggacttc aagagcaaca      360

gtgctgtggc ctggagcaac aaatctgact ttgcatgtgc aaacgccttc aacaacagca      420

ttattccaga agacaccttc ttccccagcc caggtaaggg cagctttggt gccttcgcag      480

gctgtttcct tgcttcagga atggccaggt tctgcccaga gctctggtca atgatgtcta      540

aaactcctct gattggtggt ctcggcctta tccattgcca ccaaaaccct ctttttacta      600

agaaacagtg agccttgttc tggcagtcca                                       630


<210>  19
<211>  90
<212>  PRT
<213>  Homo sapiens

<400>  19

Ile Gln Asn Pro Asp Pro Ala Val Tyr Gln Leu Arg Asp Ser Lys Ser 
1               5                   10                  15      


Ser Asp Lys Ser Val Cys Leu Phe Thr Asp Phe Asp Ser Gln Thr Asn 
            20                  25                  30          


Val Ser Gln Ser Lys Asp Ser Asp Val Tyr Ile Thr Asp Lys Thr Val 
        35                  40                  45              


Leu Asp Met Arg Ser Met Asp Phe Lys Ser Asn Ser Ala Val Ala Trp 
    50                  55                  60                  


Ser Asn Lys Ser Asp Phe Ala Cys Ala Asn Ala Phe Asn Asn Ser Ile 
65                  70                  75                  80  


Ile Pro Glu Asp Thr Phe Phe Pro Ser Pro 
                85                  90  


