                         SEQUENCE LISTING

<110>  ExcellGene SA
       WURM, Maria J.
       WURM, Florian M.
       JANCIAUSKINE, Sabina
       HAMACHER, Jurg
       STAMMBERGER, Uz
 
<120>  Methods for producing recombinant glycosylated human alpha-1 
       antitrypsin (AAT) in mammalian host cells

<130>  178225.10000

<160>  18    

<170>  PatentIn version 3.5

<210>  1
<211>  394
<212>  PRT
<213>  Homo sapiens

<400>  1

Glu Asp Pro Gln Gly Asp Ala Ala Gln Lys Thr Asp Thr Ser His His 
1               5                   10                  15      


Asp Gln Asp His Pro Thr Phe Asn Lys Ile Thr Pro Asn Leu Ala Glu 
            20                  25                  30          


Phe Ala Phe Ser Leu Tyr Arg Gln Leu Ala His Gln Ser Asn Ser Thr 
        35                  40                  45              


Asn Ile Phe Phe Ser Pro Val Ser Ile Ala Thr Ala Phe Ala Met Leu 
    50                  55                  60                  


Ser Leu Gly Thr Lys Ala Asp Thr His Asp Glu Ile Leu Glu Gly Leu 
65                  70                  75                  80  


Asn Phe Asn Leu Thr Glu Ile Pro Glu Ala Gln Ile His Glu Gly Phe 
                85                  90                  95      


Gln Glu Leu Leu Arg Thr Leu Asn Gln Pro Asp Ser Gln Leu Gln Leu 
            100                 105                 110         


Thr Thr Gly Asn Gly Leu Phe Leu Ser Glu Gly Leu Lys Leu Val Asp 
        115                 120                 125             


Lys Phe Leu Glu Asp Val Lys Lys Leu Tyr His Ser Glu Ala Phe Thr 
    130                 135                 140                 


Val Asn Phe Gly Asp Thr Glu Glu Ala Lys Lys Gln Ile Asn Asp Tyr 
145                 150                 155                 160 


Val Glu Lys Gly Thr Gln Gly Lys Ile Val Asp Leu Val Lys Glu Leu 
                165                 170                 175     


Asp Arg Asp Thr Val Phe Ala Leu Val Asn Tyr Ile Phe Phe Lys Gly 
            180                 185                 190         


Lys Trp Glu Arg Pro Phe Glu Val Lys Asp Thr Glu Glu Glu Asp Phe 
        195                 200                 205             


His Val Asp Gln Val Thr Thr Val Lys Val Pro Met Met Lys Arg Leu 
    210                 215                 220                 


Gly Met Phe Asn Ile Gln His Cys Lys Lys Leu Ser Ser Trp Val Leu 
225                 230                 235                 240 


Leu Met Lys Tyr Leu Gly Asn Ala Thr Ala Ile Phe Phe Leu Pro Asp 
                245                 250                 255     


Glu Gly Lys Leu Gln His Leu Glu Asn Glu Leu Thr His Asp Ile Ile 
            260                 265                 270         


Thr Lys Phe Leu Glu Asn Glu Asp Arg Arg Ser Ala Ser Leu His Leu 
        275                 280                 285             


Pro Lys Leu Ser Ile Thr Gly Thr Tyr Asp Leu Lys Ser Val Leu Gly 
    290                 295                 300                 


Gln Leu Gly Ile Thr Lys Val Phe Ser Asn Gly Ala Asp Leu Ser Gly 
305                 310                 315                 320 


Val Thr Glu Glu Ala Pro Leu Lys Leu Ser Lys Ala Val His Lys Ala 
                325                 330                 335     


Val Leu Thr Ile Asp Glu Lys Gly Thr Glu Ala Ala Gly Ala Met Phe 
            340                 345                 350         


Leu Glu Ala Ile Pro Met Ser Ile Pro Pro Glu Val Lys Phe Asn Lys 
        355                 360                 365             


Pro Phe Val Phe Leu Met Ile Glu Gln Asn Thr Lys Ser Pro Leu Phe 
    370                 375                 380                 


Met Gly Lys Val Val Asn Pro Thr Gln Lys 
385                 390                 


<210>  2
<211>  1210
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  Restriction Enzyme Recognition Sequence

<220>
<221>  misc_feature
<222>  (1205)..(1210)
<223>  Restriction Enzyme Recognition Sequence

<400>  2
actagtcacc gaagatcctc aaggtgacgc cgcccaaaag accgatacct cgcatcatga       60

ccaagaccac ccgaccttta acaagatcac tccaaacctg gccgagttcg cattctccct      120

ctacagacag ctggctcacc agtcaaactc aaccaacatc ttcttctccc ctgtgagcat      180

cgccactgcg ttcgccatgc tttcactggg caccaaagcc gatacgcacg acgagatcct      240

ggaggggctc aactttaacc ttaccgaaat cccggaagcg caaatccacg aaggattcca      300

agaacttctg cgcaccctca atcagccaga ctcgcagttg cagctgacta ccggcaacgg      360

actgtttctc tcggaagggc tgaaactcgt ggacaaattc ctcgaggacg tgaagaagct      420

gtaccattcg gaggcgttta ccgtcaattt cggagatacc gaagaagcta aaaagcaaat      480

caatgactac gtggagaagg gaacccaggg aaagatcgtg gacctcgtca aggaattgga      540

ccgggacacc gtgttcgccc tggtgaatta catcttcttt aaaggaaagt gggaaagacc      600

attcgaggtg aaggatactg aggaagaaga tttccacgtc gatcaggtga ctaccgtgaa      660

ggtccccatg atgaagcgcc tgggcatgtt caacatccag cactgtaaga agctgtcctc      720

gtgggtcctg ctcatgaagt acctgggaaa tgcaactgct attttcttcc tcccggatga      780

gggcaaactg cagcaccttg agaacgagct gactcatgat atcattacga agtttctgga      840

aaatgaggac aggcggagcg ccagcctcca tctcccaaag ctgtccatca cggggacgta      900

tgacctgaag tcagtccttg gacagctggg catcactaag gtgtttagca acggtgctga      960

cttgtccgga gtgactgaag aggcaccgct gaaactgtct aaggcggtcc acaaggccgt     1020

gctcaccatc gacgaaaagg gaactgaggc cgctggagca atgttcttgg aggcgatccc     1080

gatgtcgatc cctcccgaag tgaagttcaa taagccgttc gtgtttctga tgattgagca     1140

aaacactaaa agccctctgt tcatgggtaa agtggtgaac ccgactcaga agtagtgatg     1200

ataagaattc                                                            1210


<210>  3
<211>  418
<212>  PRT
<213>  Homo sapiens


<220>
<221>  SIGNAL
<222>  (1)..(24)
<223>  Natural human AAT leader sequence

<400>  3

Met Pro Ser Ser Val Ser Trp Gly Ile Leu Leu Leu Ala Gly Leu Cys 
1               5                   10                  15      


Cys Leu Val Pro Val Ser Leu Ala Glu Asp Pro Gln Gly Asp Ala Ala 
            20                  25                  30          


Gln Lys Thr Asp Thr Ser His His Asp Gln Asp His Pro Thr Phe Asn 
        35                  40                  45              


Lys Ile Thr Pro Asn Leu Ala Glu Phe Ala Phe Ser Leu Tyr Arg Gln 
    50                  55                  60                  


Leu Ala His Gln Ser Asn Ser Thr Asn Ile Phe Phe Ser Pro Val Ser 
65                  70                  75                  80  


Ile Ala Thr Ala Phe Ala Met Leu Ser Leu Gly Thr Lys Ala Asp Thr 
                85                  90                  95      


His Asp Glu Ile Leu Glu Gly Leu Asn Phe Asn Leu Thr Glu Ile Pro 
            100                 105                 110         


Glu Ala Gln Ile His Glu Gly Phe Gln Glu Leu Leu Arg Thr Leu Asn 
        115                 120                 125             


Gln Pro Asp Ser Gln Leu Gln Leu Thr Thr Gly Asn Gly Leu Phe Leu 
    130                 135                 140                 


Ser Glu Gly Leu Lys Leu Val Asp Lys Phe Leu Glu Asp Val Lys Lys 
145                 150                 155                 160 


Leu Tyr His Ser Glu Ala Phe Thr Val Asn Phe Gly Asp Thr Glu Glu 
                165                 170                 175     


Ala Lys Lys Gln Ile Asn Asp Tyr Val Glu Lys Gly Thr Gln Gly Lys 
            180                 185                 190         


Ile Val Asp Leu Val Lys Glu Leu Asp Arg Asp Thr Val Phe Ala Leu 
        195                 200                 205             


Val Asn Tyr Ile Phe Phe Lys Gly Lys Trp Glu Arg Pro Phe Glu Val 
    210                 215                 220                 


Lys Asp Thr Glu Glu Glu Asp Phe His Val Asp Gln Val Thr Thr Val 
225                 230                 235                 240 


Lys Val Pro Met Met Lys Arg Leu Gly Met Phe Asn Ile Gln His Cys 
                245                 250                 255     


Lys Lys Leu Ser Ser Trp Val Leu Leu Met Lys Tyr Leu Gly Asn Ala 
            260                 265                 270         


Thr Ala Ile Phe Phe Leu Pro Asp Glu Gly Lys Leu Gln His Leu Glu 
        275                 280                 285             


Asn Glu Leu Thr His Asp Ile Ile Thr Lys Phe Leu Glu Asn Glu Asp 
    290                 295                 300                 


Arg Arg Ser Ala Ser Leu His Leu Pro Lys Leu Ser Ile Thr Gly Thr 
305                 310                 315                 320 


Tyr Asp Leu Lys Ser Val Leu Gly Gln Leu Gly Ile Thr Lys Val Phe 
                325                 330                 335     


Ser Asn Gly Ala Asp Leu Ser Gly Val Thr Glu Glu Ala Pro Leu Lys 
            340                 345                 350         


Leu Ser Lys Ala Val His Lys Ala Val Leu Thr Ile Asp Glu Lys Gly 
        355                 360                 365             


Thr Glu Ala Ala Gly Ala Met Phe Leu Glu Ala Ile Pro Met Ser Ile 
    370                 375                 380                 


Pro Pro Glu Val Lys Phe Asn Lys Pro Phe Val Phe Leu Met Ile Glu 
385                 390                 395                 400 


Gln Asn Thr Lys Ser Pro Leu Phe Met Gly Lys Val Val Asn Pro Thr 
                405                 410                 415     


Gln Lys 
        


<210>  4
<211>  1282
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  Restriction Enzyme Recognition Site

<220>
<221>  misc_feature
<222>  (1277)..(1282)
<223>  Restriction Enzyme Recognition Site

<400>  4
actagtcacc atgccgagca gcgtgagctg gggcattctg ctgctggcgg gcctgtgctg       60

cctggtgccg gtgagcctgg cggaagatcc tcaaggtgac gccgcccaaa agaccgatac      120

ctcgcatcat gaccaagacc acccgacctt taacaagatc actccaaacc tggccgagtt      180

cgcattctcc ctctacagac agctggctca ccagtcaaac tcaaccaaca tcttcttctc      240

ccctgtgagc atcgccactg cgttcgccat gctttcactg ggcaccaaag ccgatacgca      300

cgacgagatc ctggaggggc tcaactttaa ccttaccgaa atcccggaag cgcaaatcca      360

cgaaggattc caagaacttc tgcgcaccct caatcagcca gactcgcagt tgcagctgac      420

taccggcaac ggactgtttc tctcggaagg gctgaaactc gtggacaaat tcctcgagga      480

cgtgaagaag ctgtaccatt cggaggcgtt taccgtcaat ttcggagata ccgaagaagc      540

taaaaagcaa atcaatgact acgtggagaa gggaacccag ggaaagatcg tggacctcgt      600

caaggaattg gaccgggaca ccgtgttcgc cctggtgaat tacatcttct ttaaaggaaa      660

gtgggaaaga ccattcgagg tgaaggatac tgaggaagaa gatttccacg tcgatcaggt      720

gactaccgtg aaggtcccca tgatgaagcg cctgggcatg ttcaacatcc agcactgtaa      780

gaagctgtcc tcgtgggtcc tgctcatgaa gtacctggga aatgcaactg ctattttctt      840

cctcccggat gagggcaaac tgcagcacct tgagaacgag ctgactcatg atatcattac      900

gaagtttctg gaaaatgagg acaggcggag cgccagcctc catctcccaa agctgtccat      960

cacggggacg tatgacctga agtcagtcct tggacagctg ggcatcacta aggtgtttag     1020

caacggtgct gacttgtccg gagtgactga agaggcaccg ctgaaactgt ctaaggcggt     1080

ccacaaggcc gtgctcacca tcgacgaaaa gggaactgag gccgctggag caatgttctt     1140

ggaggcgatc ccgatgtcga tccctcccga agtgaagttc aataagccgt tcgtgtttct     1200

gatgattgag caaaacacta aaagccctct gttcatgggt aaagtggtga acccgactca     1260

gaagtagtga tgataagaat tc                                              1282


<210>  5
<211>  413
<212>  PRT
<213>  Homo sapiens


<220>
<221>  SIGNAL
<222>  (1)..(19)
<223>  Human IgG heavy chain leader sequence

<400>  5

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


Val Gln Cys Glu Asp Pro Gln Gly Asp Ala Ala Gln Lys Thr Asp Thr 
            20                  25                  30          


Ser His His Asp Gln Asp His Pro Thr Phe Asn Lys Ile Thr Pro Asn 
        35                  40                  45              


Leu Ala Glu Phe Ala Phe Ser Leu Tyr Arg Gln Leu Ala His Gln Ser 
    50                  55                  60                  


Asn Ser Thr Asn Ile Phe Phe Ser Pro Val Ser Ile Ala Thr Ala Phe 
65                  70                  75                  80  


Ala Met Leu Ser Leu Gly Thr Lys Ala Asp Thr His Asp Glu Ile Leu 
                85                  90                  95      


Glu Gly Leu Asn Phe Asn Leu Thr Glu Ile Pro Glu Ala Gln Ile His 
            100                 105                 110         


Glu Gly Phe Gln Glu Leu Leu Arg Thr Leu Asn Gln Pro Asp Ser Gln 
        115                 120                 125             


Leu Gln Leu Thr Thr Gly Asn Gly Leu Phe Leu Ser Glu Gly Leu Lys 
    130                 135                 140                 


Leu Val Asp Lys Phe Leu Glu Asp Val Lys Lys Leu Tyr His Ser Glu 
145                 150                 155                 160 


Ala Phe Thr Val Asn Phe Gly Asp Thr Glu Glu Ala Lys Lys Gln Ile 
                165                 170                 175     


Asn Asp Tyr Val Glu Lys Gly Thr Gln Gly Lys Ile Val Asp Leu Val 
            180                 185                 190         


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


Phe Lys Gly Lys Trp Glu Arg Pro Phe Glu Val Lys Asp Thr Glu Glu 
    210                 215                 220                 


Glu Asp Phe His Val Asp Gln Val Thr Thr Val Lys Val Pro Met Met 
225                 230                 235                 240 


Lys Arg Leu Gly Met Phe Asn Ile Gln His Cys Lys Lys Leu Ser Ser 
                245                 250                 255     


Trp Val Leu Leu Met Lys Tyr Leu Gly Asn Ala Thr Ala Ile Phe Phe 
            260                 265                 270         


Leu Pro Asp Glu Gly Lys Leu Gln His Leu Glu Asn Glu Leu Thr His 
        275                 280                 285             


Asp Ile Ile Thr Lys Phe Leu Glu Asn Glu Asp Arg Arg Ser Ala Ser 
    290                 295                 300                 


Leu His Leu Pro Lys Leu Ser Ile Thr Gly Thr Tyr Asp Leu Lys Ser 
305                 310                 315                 320 


Val Leu Gly Gln Leu Gly Ile Thr Lys Val Phe Ser Asn Gly Ala Asp 
                325                 330                 335     


Leu Ser Gly Val Thr Glu Glu Ala Pro Leu Lys Leu Ser Lys Ala Val 
            340                 345                 350         


His Lys Ala Val Leu Thr Ile Asp Glu Lys Gly Thr Glu Ala Ala Gly 
        355                 360                 365             


Ala Met Phe Leu Glu Ala Ile Pro Met Ser Ile Pro Pro Glu Val Lys 
    370                 375                 380                 


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


Pro Leu Phe Met Gly Lys Val Val Asn Pro Thr Gln Lys 
                405                 410             


<210>  6
<211>  1267
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  Restriction Enzyme Recognition Sequence

<220>
<221>  misc_feature
<222>  (1262)..(1267)
<223>  Restriction Enzyme Recognition Sequence

<400>  6
actagtcacc atggaatttt ggctgtcctg ggttttcctc gttgcaatct tgaaaggcgt       60

ccagtgcgaa gatcctcaag gtgacgccgc ccaaaagacc gatacctcgc atcatgacca      120

agaccacccg acctttaaca agatcactcc aaacctggcc gagttcgcat tctccctcta      180

cagacagctg gctcaccagt caaactcaac caacatcttc ttctcccctg tgagcatcgc      240

cactgcgttc gccatgcttt cactgggcac caaagccgat acgcacgacg agatcctgga      300

ggggctcaac tttaacctta ccgaaatccc ggaagcgcaa atccacgaag gattccaaga      360

acttctgcgc accctcaatc agccagactc gcagttgcag ctgactaccg gcaacggact      420

gtttctctcg gaagggctga aactcgtgga caaattcctc gaggacgtga agaagctgta      480

ccattcggag gcgtttaccg tcaatttcgg agataccgaa gaagctaaaa agcaaatcaa      540

tgactacgtg gagaagggaa cccagggaaa gatcgtggac ctcgtcaagg aattggaccg      600

ggacaccgtg ttcgccctgg tgaattacat cttctttaaa ggaaagtggg aaagaccatt      660

cgaggtgaag gatactgagg aagaagattt ccacgtcgat caggtgacta ccgtgaaggt      720

ccccatgatg aagcgcctgg gcatgttcaa catccagcac tgtaagaagc tgtcctcgtg      780

ggtcctgctc atgaagtacc tgggaaatgc aactgctatt ttcttcctcc cggatgaggg      840

caaactgcag caccttgaga acgagctgac tcatgatatc attacgaagt ttctggaaaa      900

tgaggacagg cggagcgcca gcctccatct cccaaagctg tccatcacgg ggacgtatga      960

cctgaagtca gtccttggac agctgggcat cactaaggtg tttagcaacg gtgctgactt     1020

gtccggagtg actgaagagg caccgctgaa actgtctaag gcggtccaca aggccgtgct     1080

caccatcgac gaaaagggaa ctgaggccgc tggagcaatg ttcttggagg cgatcccgat     1140

gtcgatccct cccgaagtga agttcaataa gccgttcgtg tttctgatga ttgagcaaaa     1200

cactaaaagc cctctgttca tgggtaaagt ggtgaacccg actcagaagt agtgatgata     1260

agaattc                                                               1267


<210>  7
<211>  418
<212>  PRT
<213>  Homo sapiens


<220>
<221>  SIGNAL
<222>  (1)..(24)
<223>  Chimpanzee AAT leader sequence

<400>  7

Met Leu Ser Ser Val Ser Trp Gly Ile Leu Leu Leu Ala Gly Leu Cys 
1               5                   10                  15      


Cys Leu Val Pro Val Ser Leu Ala Glu Asp Pro Gln Gly Asp Ala Ala 
            20                  25                  30          


Gln Lys Thr Asp Thr Ser His His Asp Gln Asp His Pro Thr Phe Asn 
        35                  40                  45              


Lys Ile Thr Pro Asn Leu Ala Glu Phe Ala Phe Ser Leu Tyr Arg Gln 
    50                  55                  60                  


Leu Ala His Gln Ser Asn Ser Thr Asn Ile Phe Phe Ser Pro Val Ser 
65                  70                  75                  80  


Ile Ala Thr Ala Phe Ala Met Leu Ser Leu Gly Thr Lys Ala Asp Thr 
                85                  90                  95      


His Asp Glu Ile Leu Glu Gly Leu Asn Phe Asn Leu Thr Glu Ile Pro 
            100                 105                 110         


Glu Ala Gln Ile His Glu Gly Phe Gln Glu Leu Leu Arg Thr Leu Asn 
        115                 120                 125             


Gln Pro Asp Ser Gln Leu Gln Leu Thr Thr Gly Asn Gly Leu Phe Leu 
    130                 135                 140                 


Ser Glu Gly Leu Lys Leu Val Asp Lys Phe Leu Glu Asp Val Lys Lys 
145                 150                 155                 160 


Leu Tyr His Ser Glu Ala Phe Thr Val Asn Phe Gly Asp Thr Glu Glu 
                165                 170                 175     


Ala Lys Lys Gln Ile Asn Asp Tyr Val Glu Lys Gly Thr Gln Gly Lys 
            180                 185                 190         


Ile Val Asp Leu Val Lys Glu Leu Asp Arg Asp Thr Val Phe Ala Leu 
        195                 200                 205             


Val Asn Tyr Ile Phe Phe Lys Gly Lys Trp Glu Arg Pro Phe Glu Val 
    210                 215                 220                 


Lys Asp Thr Glu Glu Glu Asp Phe His Val Asp Gln Val Thr Thr Val 
225                 230                 235                 240 


Lys Val Pro Met Met Lys Arg Leu Gly Met Phe Asn Ile Gln His Cys 
                245                 250                 255     


Lys Lys Leu Ser Ser Trp Val Leu Leu Met Lys Tyr Leu Gly Asn Ala 
            260                 265                 270         


Thr Ala Ile Phe Phe Leu Pro Asp Glu Gly Lys Leu Gln His Leu Glu 
        275                 280                 285             


Asn Glu Leu Thr His Asp Ile Ile Thr Lys Phe Leu Glu Asn Glu Asp 
    290                 295                 300                 


Arg Arg Ser Ala Ser Leu His Leu Pro Lys Leu Ser Ile Thr Gly Thr 
305                 310                 315                 320 


Tyr Asp Leu Lys Ser Val Leu Gly Gln Leu Gly Ile Thr Lys Val Phe 
                325                 330                 335     


Ser Asn Gly Ala Asp Leu Ser Gly Val Thr Glu Glu Ala Pro Leu Lys 
            340                 345                 350         


Leu Ser Lys Ala Val His Lys Ala Val Leu Thr Ile Asp Glu Lys Gly 
        355                 360                 365             


Thr Glu Ala Ala Gly Ala Met Phe Leu Glu Ala Ile Pro Met Ser Ile 
    370                 375                 380                 


Pro Pro Glu Val Lys Phe Asn Lys Pro Phe Val Phe Leu Met Ile Glu 
385                 390                 395                 400 


Gln Asn Thr Lys Ser Pro Leu Phe Met Gly Lys Val Val Asn Pro Thr 
                405                 410                 415     


Gln Lys 
        


<210>  8
<211>  1282
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  Restriction Enzyme Recognition Sequence

<220>
<221>  misc_feature
<222>  (1277)..(1282)
<223>  Restriction Enzyme Recognition Sequence

<400>  8
actagtcacc atgctgagca gcgtgagctg gggcattctg ctgctggcgg gcctgtgctg       60

cctggtgccg gtgagcctgg cggaagatcc tcaaggtgac gccgcccaaa agaccgatac      120

ctcgcatcat gaccaagacc acccgacctt taacaagatc actccaaacc tggccgagtt      180

cgcattctcc ctctacagac agctggctca ccagtcaaac tcaaccaaca tcttcttctc      240

ccctgtgagc atcgccactg cgttcgccat gctttcactg ggcaccaaag ccgatacgca      300

cgacgagatc ctggaggggc tcaactttaa ccttaccgaa atcccggaag cgcaaatcca      360

cgaaggattc caagaacttc tgcgcaccct caatcagcca gactcgcagt tgcagctgac      420

taccggcaac ggactgtttc tctcggaagg gctgaaactc gtggacaaat tcctcgagga      480

cgtgaagaag ctgtaccatt cggaggcgtt taccgtcaat ttcggagata ccgaagaagc      540

taaaaagcaa atcaatgact acgtggagaa gggaacccag ggaaagatcg tggacctcgt      600

caaggaattg gaccgggaca ccgtgttcgc cctggtgaat tacatcttct ttaaaggaaa      660

gtgggaaaga ccattcgagg tgaaggatac tgaggaagaa gatttccacg tcgatcaggt      720

gactaccgtg aaggtcccca tgatgaagcg cctgggcatg ttcaacatcc agcactgtaa      780

gaagctgtcc tcgtgggtcc tgctcatgaa gtacctggga aatgcaactg ctattttctt      840

cctcccggat gagggcaaac tgcagcacct tgagaacgag ctgactcatg atatcattac      900

gaagtttctg gaaaatgagg acaggcggag cgccagcctc catctcccaa agctgtccat      960

cacggggacg tatgacctga agtcagtcct tggacagctg ggcatcacta aggtgtttag     1020

caacggtgct gacttgtccg gagtgactga agaggcaccg ctgaaactgt ctaaggcggt     1080

ccacaaggcc gtgctcacca tcgacgaaaa gggaactgag gccgctggag caatgttctt     1140

ggaggcgatc ccgatgtcga tccctcccga agtgaagttc aataagccgt tcgtgtttct     1200

gatgattgag caaaacacta aaagccctct gttcatgggt aaagtggtga acccgactca     1260

gaagtagtga tgataagaat tc                                              1282


<210>  9
<211>  6504
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1948)..(1953)
<223>  Restriction Enzyme Recognition Sequence

<220>
<221>  misc_feature
<222>  (2015)..(3196)
<223>  CHO-cell codon-optimized AAT protein sequence

<220>
<221>  misc_feature
<222>  (3209)..(3214)
<223>  Restriction Enzyme Recognition Sequence

<400>  9
ggcgcgcctt aaccctagaa agatagtctg cgtaaaattg acgcatgcat tcttgaaata       60

ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc      120

cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta      180

taacgaccgc gtgagtcaaa atgacgcatg attatctttt acgtgacttt taagatttaa      240

ctcatacgat aattatattg ttatttcatg ttctacttac gtgataactt attatatata      300

tattttcttg ttatagatat catcgataac aggaaagttc cattggagcc aagtacattg      360

agtcaatagg gactttccaa tgggttttgc ccagtacata aggtcaatgg gaggtaagcc      420

aatgggtttt tcccattact ggcacgtata ctgagtcatt agggactttc caatgggttt      480

tgcccagtac ataaggtcaa taggggtgaa tcaacaggaa agttccattg gagccaagta      540

cactgagtca atagggactt tccattgggt tttgcccagt acaaaaggtc aatagggggt      600

gagtcaatgg gtttttccca ttattggcac gtacataagg tcaatagggg tgagtcattg      660

ggtttttcca gccaatttaa ttaaaacgcc atgtactttc ccaccattga cgtcaatggg      720

ctattgaaac taatgcaacg tgacctttaa acggtacttt cccatagctg attaatggga      780

aagtaccgtt ctcgagccaa tacacgtcaa tgggaagtga aagggcagcc aaaacgtaac      840

accgccccgg ttttcccctg gaaattccat attggcacgc attctattgg ctgagctgcg      900

ttctacgtgg gtataagagg cgcgaccagc gtcggtaccg tcgcagtctt cggtctgacc      960

accgtagaac gcagagctcc tcgctgcagg caagcttggt aagtgccgtg tgtggttccc     1020

gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacgcccc     1080

tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg     1140

aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc     1200

tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag     1260

tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt     1320

cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc     1380

gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca     1440

ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg     1500

ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga     1560

gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc     1620

tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc     1680

gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg     1740

agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc     1800

cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg     1860

gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa     1920

agtttttttc ttccatttca gggatccact agtcaccatg gaattttggc tgtcctgggt     1980

tttcctcgtt gcaatcttga aaggcgtcca gtgcgaagat cctcaaggtg acgccgccca     2040

aaagaccgat acctcgcatc atgaccaaga ccacccgacc tttaacaaga tcactccaaa     2100

cctggccgag ttcgcattct ccctctacag acagctggct caccagtcaa actcaaccaa     2160

catcttcttc tcccctgtga gcatcgccac tgcgttcgcc atgctttcac tgggcaccaa     2220

agccgatacg cacgacgaga tcctggaggg gctcaacttt aaccttaccg aaatcccgga     2280

agcgcaaatc cacgaaggat tccaagaact tctgcgcacc ctcaatcagc cagactcgca     2340

gttgcagctg actaccggca acggactgtt tctctcggaa gggctgaaac tcgtggacaa     2400

attcctcgag gacgtgaaga agctgtacca ttcggaggcg tttaccgtca atttcggaga     2460

taccgaagaa gctaaaaagc aaatcaatga ctacgtggag aagggaaccc agggaaagat     2520

cgtggacctc gtcaaggaat tggaccggga caccgtgttc gccctggtga attacatctt     2580

ctttaaagga aagtgggaaa gaccattcga ggtgaaggat actgaggaag aagatttcca     2640

cgtcgatcag gtgactaccg tgaaggtccc catgatgaag cgcctgggca tgttcaacat     2700

ccagcactgt aagaagctgt cctcgtgggt cctgctcatg aagtacctgg gaaatgcaac     2760

tgctattttc ttcctcccgg atgagggcaa actgcagcac cttgagaacg agctgactca     2820

tgatatcatt acgaagtttc tggaaaatga ggacaggcgg agcgccagcc tccatctccc     2880

aaagctgtcc atcacgggga cgtatgacct gaagtcagtc cttggacagc tgggcatcac     2940

taaggtgttt agcaacggtg ctgacttgtc cggagtgact gaagaggcac cgctgaaact     3000

gtctaaggcg gtccacaagg ccgtgctcac catcgacgaa aagggaactg aggccgctgg     3060

agcaatgttc ttggaggcga tcccgatgtc gatccctccc gaagtgaagt tcaataagcc     3120

gttcgtgttt ctgatgattg agcaaaacac taaaagccct ctgttcatgg gtaaagtggt     3180

gaacccgact cagaagtagt gatgataaga attctgcaga tatccatcac actggcggcc     3240

gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct     3300

gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc     3360

cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg     3420

catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca     3480

agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt     3540

ctgaggcgga aagaaccagt ggcggtaata cggttatcca cagaatcagg ggataacgca     3600

ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg     3660

ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt     3720

cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc     3780

ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct     3840

tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc     3900

gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta     3960

tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca     4020

gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag     4080

tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag     4140

ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt     4200

agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa     4260

gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg     4320

attttggtca taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca     4380

caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca     4440

tcaatgtatc ttatcatgtc tggatccgct tcaggcaccg ggcttgcggg tcatgcacca     4500

ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta     4560

gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc     4620

cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga     4680

gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc     4740

ttccatctgt tgctgcgcgg ccagcctgga accgctcaac tcggccatgc gcgggccgat     4800

ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc     4860

gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc     4920

ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc gcgtggcggg     4980

gtagtcggcg aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt cgcgggtggc     5040

gaggcgcacc gtgggcttgt actcggtcat ggtggcctgc agagtcgctc tgtgttcgag     5100

gccacacgcg tcaccttaat atgcgaagtg gacctgggac cgcgccgccc cgactgcatc     5160

tgcgtgtttt cgccaatgac aagacgctgg gcggggtttg tgtcatcata gaactaaaga     5220

catgcaaata tatttcttcc ggggacaccg ccagcaaacg cgagcaacgg gccacgggga     5280

tgaagcagct ggctagctaa aagttttgtt actttataga agaaattttg agtttttgtt     5340

tttttttaat aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta     5400

agtgtaaata taataaaact taatatctat tcaaattaat aaataaacct cgatatacag     5460

accgataaaa cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg     5520

attatctttc tagggttaat tcgaacagct ggttctttcc gcctcaggac tcttcctttt     5580

tcaataaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa     5640

tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc     5700

ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga     5760

taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa     5820

gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt     5880

gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg     5940

ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc     6000

aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg     6060

gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag     6120

cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt     6180

actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt     6240

caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac     6300

gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac     6360

ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag     6420

caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa     6480

tactcatact cttccttttt caat                                            6504


<210>  10
<211>  594
<212>  PRT
<213>  Trichoplusia ni

<400>  10

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


Ser Asp Asp Glu Leu Val Gly Glu Asp Ser Asp Ser Glu Ile Ser Asp 
            20                  25                  30          


His Val Ser Glu Asp Asp Val Gln Ser Asp Thr Glu Glu Ala Phe Ile 
        35                  40                  45              


Asp Glu Val His Glu Val Gln Pro Thr Ser Ser Gly Ser Glu Ile Leu 
    50                  55                  60                  


Asp Glu Gln Asn Val Ile Glu Gln Pro Gly Ser Ser Leu Ala Ser Asn 
65                  70                  75                  80  


Arg Ile Leu Thr Leu Pro Gln Arg Thr Ile Arg Gly Lys Asn Lys His 
                85                  90                  95      


Cys Trp Ser Thr Ser Lys Ser Thr Arg Arg Ser Arg Val Ser Ala Leu 
            100                 105                 110         


Asn Ile Val Arg Ser Gln Arg Gly Pro Thr Arg Met Cys Arg Asn Ile 
        115                 120                 125             


Tyr Asp Pro Leu Leu Cys Phe Lys Leu Phe Phe Thr Asp Glu Ile Ile 
    130                 135                 140                 


Ser Glu Ile Val Lys Trp Thr Asn Ala Glu Ile Ser Leu Lys Arg Arg 
145                 150                 155                 160 


Glu Ser Met Thr Gly Ala Thr Phe Arg Asp Thr Asn Glu Asp Glu Ile 
                165                 170                 175     


Tyr Ala Phe Phe Gly Ile Leu Val Met Thr Ala Val Arg Lys Asp Asn 
            180                 185                 190         


His Met Ser Thr Asp Asp Leu Phe Asp Arg Ser Leu Ser Met Val Tyr 
        195                 200                 205             


Val Ser Val Met Ser Arg Asp Arg Phe Asp Phe Leu Ile Arg Cys Leu 
    210                 215                 220                 


Arg Met Asp Asp Lys Ser Ile Arg Pro Thr Leu Arg Glu Asn Asp Val 
225                 230                 235                 240 


Phe Thr Pro Val Arg Lys Ile Trp Asp Leu Phe Ile His Gln Cys Ile 
                245                 250                 255     


Gln Asn Tyr Thr Pro Gly Ala His Leu Thr Ile Asp Glu Gln Leu Leu 
            260                 265                 270         


Gly Phe Arg Gly Arg Cys Pro Phe Arg Met Tyr Ile Pro Asn Lys Pro 
        275                 280                 285             


Ser Lys Tyr Gly Ile Lys Ile Leu Met Met Cys Asp Ser Gly Thr Lys 
    290                 295                 300                 


Tyr Met Ile Asn Gly Met Pro Tyr Leu Gly Arg Gly Thr Gln Thr Asn 
305                 310                 315                 320 


Gly Val Pro Leu Gly Glu Tyr Tyr Val Lys Glu Leu Ser Lys Pro Val 
                325                 330                 335     


His Gly Ser Cys Arg Asn Ile Thr Cys Asp Asn Trp Phe Thr Ser Ile 
            340                 345                 350         


Pro Leu Ala Lys Asn Leu Leu Gln Glu Pro Tyr Lys Leu Thr Ile Val 
        355                 360                 365             


Gly Thr Val Arg Ser Asn Lys Arg Glu Ile Pro Glu Val Leu Lys Asn 
    370                 375                 380                 


Ser Arg Ser Arg Pro Val Gly Thr Ser Met Phe Cys Phe Asp Gly Pro 
385                 390                 395                 400 


Leu Thr Leu Val Ser Tyr Lys Pro Lys Pro Ala Lys Met Val Tyr Leu 
                405                 410                 415     


Leu Ser Ser Cys Asp Glu Asp Ala Ser Ile Asn Glu Ser Thr Gly Lys 
            420                 425                 430         


Pro Gln Met Val Met Tyr Tyr Asn Gln Thr Lys Gly Gly Val Asp Thr 
        435                 440                 445             


Leu Asp Gln Met Cys Ser Val Met Thr Cys Ser Arg Lys Thr Asn Arg 
    450                 455                 460                 


Trp Pro Met Ala Leu Leu Tyr Gly Met Ile Asn Ile Ala Cys Ile Asn 
465                 470                 475                 480 


Ser Phe Ile Ile Tyr Ser His Asn Val Ser Ser Lys Gly Glu Lys Val 
                485                 490                 495     


Gln Ser Arg Lys Lys Phe Met Arg Asn Leu Tyr Met Ser Leu Thr Ser 
            500                 505                 510         


Ser Phe Met Arg Lys Arg Leu Glu Ala Pro Thr Leu Lys Arg Tyr Leu 
        515                 520                 525             


Arg Asp Asn Ile Ser Asn Ile Leu Pro Asn Glu Val Pro Gly Thr Ser 
    530                 535                 540                 


Asp Asp Ser Thr Glu Glu Pro Val Met Lys Lys Arg Thr Tyr Cys Thr 
545                 550                 555                 560 


Tyr Cys Pro Ser Lys Ile Arg Arg Lys Ala Asn Ala Ser Cys Lys Lys 
                565                 570                 575     


Cys Lys Lys Val Ile Cys Arg Glu His Asn Ile Asp Met Cys Gln Ser 
            580                 585                 590         


Cys Phe 
        


<210>  11
<211>  24
<212>  PRT
<213>  Homo sapiens


<220>
<221>  SIGNAL
<222>  (1)..(24)
<223>  H. sapiens & Hylobates sp. natural AAT leader sequence

<400>  11

Met Pro Ser Ser Val Ser Trp Gly Ile Leu Leu Leu Ala Gly Leu Cys 
1               5                   10                  15      


Cys Leu Val Pro Val Ser Leu Ala 
            20                  


<210>  12
<211>  19
<212>  PRT
<213>  Homo sapiens


<220>
<221>  Signal
<222>  (1)..(19)
<223>  H. sapiens IgG Heavy chain leader sequence

<400>  12

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


Val Gln Cys 
            


<210>  13
<211>  24
<212>  PRT
<213>  Pan troglodytes


<220>
<221>  Signal
<222>  (1)..(24)
<223>  Pan troglodytes (Chimpanzee) AAT leader sequence

<400>  13

Met Leu Ser Ser Val Ser Trp Gly Ile Leu Leu Leu Ala Gly Leu Cys 
1               5                   10                  15      


Cys Leu Val Pro Val Ser Leu Ala 
            20                  


<210>  14
<211>  18
<212>  PRT
<213>  Homo sapiens


<220>
<221>  Signal
<222>  (1)..(18)
<223>  H. sapiens serum albumin leader sequence

<400>  14

Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ala 
1               5                   10                  15      


Tyr Ser 
        


<210>  15
<211>  19
<212>  PRT
<213>  Homo sapiens


<220>
<221>  Signal
<222>  (1)..(19)
<223>  H. sapiens Azurocidin leader sequence

<400>  15

Met Thr Arg Leu Thr Val Leu Ala Leu Leu Ala Gly Leu Leu Ala Ser 
1               5                   10                  15      


Ser Arg Ala 
            


<210>  16
<211>  22
<212>  PRT
<213>  Homo sapiens


<220>
<221>  Signal
<222>  (1)..(22)
<223>  H. sapiens Ig kappa Light chain leader sequence

<400>  16

Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro 
1               5                   10                  15      


Val Ser Asp Thr Thr Gly 
            20          


<210>  17
<211>  20
<212>  PRT
<213>  Mus musculus


<220>
<221>  Signal
<222>  (1)..(20)
<223>  Mouse and Hamster Ig kappa Light chain

<400>  17

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 
1               5                   10                  15      


Gly Ser Thr Gly 
            20  


<210>  18
<211>  9
<212>  PRT
<213>  Synthetic


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  (7-Methoxycoumarin-4-yl)acetyl (Mca) attached before residue 
       position 1

<220>
<221>  MOD_RES
<222>  (6)..(6)
<223>  Xaa = Norvaline (Nva)

<220>
<221>  MISC_FEATURE
<222>  (9)..(9)
<223>  2,4-Dinitrophenyl (Dnp) attached after residue position 9

<400>  18

Arg Pro Lys Val Glu Xaa Trp Arg Lys 
1               5                   


