                         SEQUENCE LISTING

<110>  Imperial College Innovations Limited
 
<120>  Methods and Polynucleotides

<130>  P141925WO

<150>  GB2106646.9
<151>  2021-05-10

<150>  GR20210100201
<151>  2021-03-29

<160>  16    

<170>  PatentIn version 3.5

<210>  1
<211>  2481
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Encodes a solubility tag, a catalytic domain of an enzyme 
       involved in a glycosylation pathway (NtGnTI), a linker and an 
       AviTag.

<400>  1
atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt       60

ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat      120

ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt      180

atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc      240

accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac      300

aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa      360

gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg      420

aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg      480

ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa      540

gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt      600

aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa      660

ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa      720

gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt      780

ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc      840

ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg      900

ggtgccgtag cgctgaagtc ttacgaggaa gagttggtga aagatccgcg tattgccgcc      960

actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc     1020

tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa     1080

gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac     1140

aacctcggga tcgagggaag gatttcacat atggcgacgc aatcggaata tgcggatcgc     1200

ttggcagcgg cgattgaagc cgaaaaccat tgcacgtcac agacccgctt actgatcgat     1260

caaatcagtc agcaacaagg acgcattgta gccctggagg aacagatgaa acgtcaggat     1320

caggagtgtc gccaattacg tgctttggtt caggatcttg agtcgaaagg gattaagaaa     1380

ttgatcggca atgttcaaat gcctgttgct gcagtagtgg tcatggcctg caatcgtgcg     1440

gattacctcg agaaaaccat caaatccatc ctgaagtatc agattagcgt tgcaccgaaa     1500

taccctctgt ttatctccca agatggttct catccggatg tccgcaaact ggcgttaagc     1560

tacgatcaac tgacctatat gcagcatctg gattttgaac cggtgcacac tgaacgtcct     1620

ggcgaattaa tcgcgtatta caaaattgca cgccactaca aatgggccct tgaccagctc     1680

ttttacaagc acaactttag ccgggtgatc attcttgagg acgatatgga aattgcccca     1740

gacttcttcg acttctttga agccggagct actctgctgg atcgcgataa gtcgattatg     1800

gcgatcagta gctggaacga taacgggcag atgcagtttg tgcaagatcc ctatgcttta     1860

tatcgctcag acttctttcc gggtctgggt tggatgttga gtaaatcgac atgggacgaa     1920

ctgagcccga aatggccgaa agcttactgg gatgactggt tgcgcctgaa ggaaaaccat     1980

cgtggtcgtc agttcattcg cccggaagtg tgtcgtagct ataactttgg tgaacatggt     2040

agcagtctgg gccagttctt taaacagtat ctggaaccca tcaaactcaa tgacgtccag     2100

gtcgactgga aatccatgga tctttcttat ctgctggagg acaattacgt gaaacacttt     2160

ggcgatctgg tgaagaaagc gaaaccgatt catggtgccg acgcagtgct gaaagcgttt     2220

aacattgatg gggatgttcg cattcagtac cgtgatcagc tggactttga agatattgca     2280

cgtcagtttg gcattttcga agagtggaaa gatggcgtac cacgtgcggc ctataaaggc     2340

atcgtagtgt tccgctatca gacgtcacgc cgggttttcc tcgtcggccc agactctctg     2400

cagcaactgg gcaatgaaga taccgaattc ggttctggtc ttaatgatat ttttgaagct     2460

cagaagattg aatggcatga a                                               2481


<210>  2
<211>  2256
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Encodes a solubility tag, a catalytic domain of an enzyme 
       involved in a glycosylation pathway (hGnTI), a linker and an 
       AviTag.

<400>  2
atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt       60

ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat      120

ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt      180

atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc      240

accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac      300

aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa      360

gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg      420

aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg      480

ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa      540

gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt      600

aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa      660

ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa      720

gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt      780

ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc      840

ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg      900

ggtgccgtag cgctgaagtc ttacgaggaa gagttggtga aagatccgcg tattgccgcc      960

actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc     1020

tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa     1080

gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac     1140

aacctcggga tcgagggaag gatttcacat atggcggtga ttccgatcct ggtcattgcg     1200

tgtgaccgtt cgaccgtgcg tcgttgcctg gataaactgt tgcattaccg cccgtctgcc     1260

gagctgtttc caatcattgt ttctcaagac tgcggccatg aggaaaccgc tcaagcgatc     1320

gcaagctatg gtagcgcggt tacgcacatc cgccagccgg atctgtccag catcgcggtt     1380

ccgccggatc accgcaaatt ccaaggttac tacaaaattg cgcgtcatta tcgttgggcg     1440

ctgggtcagg tatttcgcca gtttcgcttt ccggcagcgg tcgtcgtcga ggatgatctg     1500

gaggttgccc cagacttctt cgagtacttc cgtgcgacgt atccgttgct gaaggcagat     1560

ccgtccctgt ggtgcgtcag cgcgtggaat gataacggta aagagcagat ggtggatgcc     1620

agccgtcctg aactgctgta ccgtaccgac ttctttccgg gcctgggttg gctgctgttg     1680

gctgaactgt gggcggaact ggagccgaag tggccgaaag cattttggga cgattggatg     1740

cgtcgcccgg aacagcgcca gggccgtgcc tgtattcgcc cggagattag ccgcaccatg     1800

acgtttggtc gcaagggcgt gagccacggc cagttctttg accagcatct gaaattcatt     1860

aagctgaatc agcaattcgt tcacttcacc caactggacc tgagctactt gcaacgtgag     1920

gcgtatgatc gtgacttctt ggcgcgtgtc tatggtgctc cgcaactgca agtcgagaaa     1980

gtgcgcacga acgatcgtaa ggagctgggt gaggtgcgcg tgcagtacac cggccgtgac     2040

agctttaagg ccttcgccaa ggcgctgggc gtcatggacg acctgaaaag cggcgttcct     2100

cgtgcgggtt atcgtggtat tgtgaccttt cagttccgtg gtcgtcgcgt tcatctggca     2160

ccgccgctga cctgggaagg ctacgacccg agctggaacg aattcggttc tggtcttaat     2220

gatatttttg aagctcagaa gattgaatgg catgaa                               2256


<210>  3
<211>  2040
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Encodes a solubility tag, a catalytic domain of an enzyme 
       involved in a glycosylation pathway (hGalT), a linker and an 
       AviTag.

<400>  3
atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt       60

ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat      120

ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt      180

atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc      240

accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac      300

aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa      360

gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg      420

aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg      480

ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa      540

gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt      600

aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa      660

ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa      720

gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt      780

ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc      840

ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg      900

ggtgccgtag cgctgaagtc ttacgaggaa gagttggtga aagatccgcg tattgccgcc      960

actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc     1020

tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa     1080

gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac     1140

aacctcggga tcgagggaag gatttcacat atggcctgcc ctgaggaaag cccactgttg     1200

gtgggcccaa tgctgatcga gtttaacatg ccggtggacc tggaactggt ggcgaaacag     1260

aacccgaacg tcaaaatggg cggccgttac gcaccgcgtg actgcgttag cccgcacaaa     1320

gtcgcgatca ttattccgtt ccgcaatcgc caagagcatc tgaagtactg gctgtactat     1380

ctgcatccag ttctgcaacg tcagcaattg gactacggta tttacgttat caatcaagcc     1440

ggcgacacga tctttaatcg tgctaagttg ctgaatgttg gttttcaaga agcgctgaaa     1500

gactacgact acacctgttt cgtgttctcc gacgttgacc tgattccgat gaatgatcac     1560

aatgcgtacc gctgtttttc tcagccgcgt cacatcagcg tagcgatgga taagtttggt     1620

ttcagcctgc cgtatgtgca gtattttggt ggcgtcagcg cactgagcaa gcaacagttt     1680

ctcacgatta acggtttccc gaacaactat tggggttggg gtggcgaaga tgatgatatc     1740

ttcaaccgtc tggtgttccg tggtatgagc attagccgcc cgaacgctgt ggttggccgt     1800

tgccgtatga ttcgtcatag ccgcgacaag aaaaatgaac cgaatcctca gcgtttcgat     1860

cgtatcgcac acaccaaaga aactatgttg agcgacggct taaacagcct gacctatcaa     1920

gtcttggatg ttcaacgcta tccgctgtac acgcagatta ccgtggacat tggcaccccg     1980

agcgaattcg gttctggtct taatgatatt tttgaagctc agaagattga atggcatgaa     2040


<210>  4
<211>  57
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward strand for encoding Avitag peptide sequence and GS 
       linker.

<400>  4
aattcggttc tggtcttaat gatatttttg aagctcagaa gattgaatgg catgaaa          57


<210>  5
<211>  57
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse strand for encoding Avitag peptide sequence and GS 
       linker.

<400>  5
agcttttcat gccattcaat cttctgagct tcaaaaatat cattaagacc agaaccg          57


<210>  6
<211>  4
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Linker for preparing recombinant fusion protein.

<400>  6

Gly Ser Gly Ser 
1               


<210>  7
<211>  6
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Linker for preparing recombinant fusion protein.

<400>  7

Gly Ser Gly Ser Gly Ser 
1               5       


<210>  8
<211>  8
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Linker for preparing recombinant fusion protein.

<400>  8

Gly Ser Gly Ser Gly Ser Gly Ser 
1               5               


<210>  9
<211>  10
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Linker for preparing recombinant fusion protein.

<400>  9

Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
1               5                   10  


<210>  10
<211>  5
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Linker for preparing recombinant fusion protein.


<220>
<221>  REPEAT
<222>  (1)..(5)
<223>  Can be repeated n times, where n is typically 1, 2 or 3.

<400>  10

Gly Gly Gly Gly Ser 
1               5   


<210>  11
<211>  5
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Linker for preparing recombinant fusion protein.


<220>
<221>  REPEAT
<222>  (1)..(5)
<223>  Can be repeated n times, where n is typically 1, 2 or 3.

<400>  11

Glu Ala Ala Ala Lys 
1               5   


<210>  12
<211>  15
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AviTag

<400>  12

Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 
1               5                   10                  15  


<210>  13
<211>  13
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AviTag


<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  X can be any amino acid.

<220>
<221>  MISC_FEATURE
<222>  (3)..(3)
<223>  X can be any amino acid except for L, V, I, W, F or Y.

<400>  13

Leu Xaa Xaa Ile Phe Glu Ala Gln Lys Ile Glu Trp Arg 
1               5                   10              


<210>  14
<211>  15
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AviTag (BioTag)

<400>  14

Ala Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Ala 
1               5                   10                  15  


<210>  15
<211>  23
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AviTag (BLRP)

<400>  15

Met Ala Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp 
1               5                   10                  15      


His Glu Asp Thr Gly Gly Ser 
            20              


<210>  16
<211>  15
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AviTag (BirA Substrate Peptide)

<400>  16

Leu His His Ile Leu Asp Ala Gln Lys Met Val Trp Asn His Arg 
1               5                   10                  15  


