                         SEQUENCE LISTING

<110>  The University of North Carolina at Chapel Hill
       Xiao, Xiao
       Li, Juan
       Yuan, Zhenhua
 
<120>  OPTIMIZED HUMAN CLOTTING FACTOR VIII GENE EXPRESSION CASSETTES 
       AND THEIR USE

<130>  5470-734WO

<150>  US 62/112,901
<151>  2015-02-06

<160>  34    

<170>  PatentIn version 3.5

<210>  1
<211>  200
<212>  DNA
<213>  Homo sapiens

<400>  1
ctgtttactc tggttaattt ttaaaggagg gtaaacagtg cctgaaagct gacctttgcc       60

cacattcctc cggtagacat taacttatta aattgattct gattacaaat ctgacctttg      120

cccccatctc acccagtaac aatgcaagag ttgatgtcag tctataaaaa gcgaagcgcg      180

cggtgggcgg ggttcgctgc                                                  200


<210>  2
<211>  288
<212>  DNA
<213>  Artificial

<220>
<223>  LXP3.3I synthetic promoter sequence

<400>  2
ctgtttactc tggttaattt ttaaaggagg gtaaacagtg cctgaaagct gacctttgcc       60

cacattcctc cggtagacat taacttatta aattgattct gattacaaat ctgacctttg      120

cccccatctc acccagtaac aatgcaagag ttgatgtcag tctataaaaa gcgaagcgcg      180

cggtgggcgg ggttcgctgc ctgcaggtga gtatctcagg gatccagaca tggggatatg      240

ggaggtgcct ctgatcccag ggctcactgt gggtctctct gttcacag                   288


<210>  3
<211>  4374
<212>  DNA
<213>  Artificial

<220>
<223>  Synthetic human B-domain deleted factor FVIII coding sequence

<400>  3
atgcagatcg agctgtctac ctgcttcttc ctgtgcctgc tgcggttctg cttcagcgcc       60

accagacggt actatctggg cgccgtggaa ctgagctggg actacatgca gagcgacctg      120

ggcgagctgc ccgtggatgc cagattccct ccaagagtgc ccaagagctt ccccttcaac      180

acctccgtgg tgtacaagaa aaccctgttc gtggaattca ccgaccacct gttcaatatc      240

gccaagccca gacccccctg gatgggcctg ctgggaccta caattcaggc cgaggtgtac      300

gacaccgtcg tgatcaccct gaagaacatg gccagccacc ccgtgtctct gcatgccgtg      360

ggagtgtcct actggaaggc ctctgagggc gccgagtacg acgatcagac cagccagcgc      420

gagaaagagg acgacaaggt gttccctggc ggcagccaca cctacgtgtg gcaggtgctg      480

aaagaaaacg gccccatggc ctccgaccct ctgtgcctga catacagcta cctgagccac      540

gtggacctcg tgaaggacct gaacagcggc ctgatcggag ccctgctcgt gtgtagagag      600

ggcagcctgg ccaaagagaa aacccagacc ctgcacaagt tcatcctgct gttcgccgtg      660

ttcgacgagg gcaagagctg gcacagcgag acaaagaaca gcctgatgca ggaccgggac      720

gccgcctctg ctagagcctg gcccaaaatg cacaccgtga acggctacgt gaacagaagc      780

ctgcccggac tgatcggctg ccaccggaag tctgtgtact ggcacgtgat cggcatgggc      840

accacccctg aggtgcacag catctttctg gaaggacaca cctttctcgt gcggaaccac      900

cggcaggcca gcctggaaat cagccctatc accttcctga ccgcccagac actgctgatg      960

gacctgggcc agtttctgct gttctgccac atcagctccc accagcacga cggcatggaa     1020

gcctacgtga aggtggacag ctgccccgag gaaccccagc tgcggatgaa gaacaacgag     1080

gaagccgagg actacgacga cgacctgacc gacagcgaga tggacgtggt gcgcttcgac     1140

gacgataaca gccccagctt catccagatc agaagcgtgg ccaagaagca ccccaagacc     1200

tgggtgcact atatcgccgc cgaggaagag gactgggatt acgcccctct ggtgctggcc     1260

cccgacgaca gaagctacaa gagccagtac ctgaacaatg gcccccagcg gatcggccgg     1320

aagtataaga aagtgcggtt catggcctac accgacgaga cattcaagac cagagaggcc     1380

atccagcacg agagcggcat cctgggccct ctgctgtatg gcgaagtggg cgacaccctg     1440

ctgatcatct tcaagaacca ggccagcaga ccctacaaca tctaccctca cggcatcacc     1500

gacgtgcggc ccctgtactc tagaaggctg cccaagggcg tgaaacacct gaaggacttc     1560

cccatcctgc ccggcgagat tttcaagtac aagtggaccg tgaccgtgga agatggcccc     1620

accaagagcg accccagatg cctgacacgg tactacagca gcttcgtgaa catggaacgg     1680

gacctggcct ccggcctgat tggcccactg ctgatctgct acaaagaaag cgtggaccag     1740

cggggcaacc agatcatgag cgacaagcgg aacgtgatcc tgtttagcgt gttcgatgag     1800

aaccggtcct ggtatctgac cgagaatatc cagcggttcc tgcccaaccc tgccggcgtg     1860

cagctggaag atcctgagtt ccaggcctcc aacatcatgc actccatcaa tggctatgtg     1920

ttcgacagcc tgcagctgag cgtgtgcctg cacgaggtgg cctactggta catcctgagc     1980

atcggggccc agaccgactt cctgtccgtg ttcttctccg gctacacctt caagcacaag     2040

atggtgtacg aggataccct gaccctgttc ccctttagcg gcgaaaccgt gttcatgagc     2100

atggaaaacc ccggcctgtg gatcctgggc tgccacaaca gcgacttccg gaacagaggc     2160

atgaccgccc tgctgaaggt gtccagctgc gacaagaaca ccggcgacta ctacgaggac     2220

agctatgagg acatcagcgc ctacctgctg agcaagaaca atgccatcga gcccagaagc     2280

ttcagccagc cccctgtgct gaagcggcac cagagagaga tcacccggac caccctgcag     2340

tccgaccagg aagagatcga ttacgacgac accatcagcg tggaaatgaa gaaagaagat     2400

ttcgacatct acgacgagga cgagaaccag agcccccggt cctttcagaa aaagacccgg     2460

cactacttca ttgccgctgt ggaacggctg tgggactacg gcatgagcag cagccctcac     2520

gtgctgagaa acagggccca gagcggcagc gtgccccagt tcaagaaagt ggtgttccag     2580

gaattcacag acggcagctt tacccagcct ctgtaccgcg gcgagctgaa cgaacacctg     2640

ggactgctgg gcccctatat ccgggccgaa gtggaagata acatcatggt caccttccgg     2700

aatcaggcct cccggcccta cagcttctac agctccctga tcagctacga agaggaccag     2760

agacagggcg ctgagccccg gaagaacttc gtgaagccca acgagactaa gacctacttt     2820

tggaaggtgc agcaccacat ggcccctaca aaggacgagt tcgactgcaa ggcctgggcc     2880

tacttctccg atgtggacct ggaaaaggac gtgcactctg ggctgatcgg ccccctgctc     2940

gtgtgccaca ccaacaccct gaatcccgcc cacggcagac aagtgacagt gcaggaattc     3000

gccctgttct tcaccatctt cgacgaaaca aagagctggt acttcaccga aaacatggaa     3060

agaaactgcc gggctccctg caacatccag atggaagatc ccaccttcaa agagaactac     3120

cggttccacg ccatcaacgg ctacatcatg gacacactgc ccggcctcgt gatggctcag     3180

gatcagcgga tccggtggta tctgctgtcc atgggctcca acgagaacat ccacagcatc     3240

cacttcagcg gccacgtgtt caccgtgcgg aaaaaagaag agtacaaaat ggccctgtac     3300

aacctgtacc ctggggtgtt cgagacagtg gaaatgctgc ccagcaaggc cggcatctgg     3360

cgggtggaat gtctgatcgg cgagcatctg cacgctggga tgagcacact gtttctggtg     3420

tacagcaaca agtgccagac acctctgggc atggcctctg gccacatccg ggactttcag     3480

atcacagcca gcggccagta tggccagtgg gccccaaaac tggccagact gcactacagc     3540

ggcagcatca acgcctggtc caccaaagag cccttcagct ggatcaaggt ggacctgctg     3600

gctcccatga tcatccacgg aatcaagacc cagggcgcca gacagaagtt ctccagcctg     3660

tacatctccc agttcatcat catgtactcc ctggacggca agaagtggca gacctaccgg     3720

ggcaatagca ccggcaccct gatggtgttc ttcggcaacg tggactccag cggcattaag     3780

cacaacatct tcaacccccc catcattgcc cggtacatcc ggctgcaccc cacccactac     3840

agcatccggt ccaccctgag aatggaactg atgggctgcg acctgaactc ctgcagcatg     3900

cccctgggga tggaaagcaa ggccatctcc gacgcccaga tcaccgcctc cagctacttc     3960

accaacatgt tcgccacctg gtccccatcc aaggcccggc tgcatctgca gggcagaagc     4020

aatgcttgga ggccccaagt gaacaacccc aaagaatggc tgcaggtgga cttccagaaa     4080

accatgaaag tgaccggcgt gaccacccag ggcgtgaagt ctctgctgac ctctatgtac     4140

gtgaaagagt tcctgatctc cagcagccag gacggccacc agtggaccct gtttttccag     4200

aacggcaaag tgaaagtgtt tcaggggaac caggacagct tcacccccgt cgtgaatagc     4260

ctggaccctc cactgctgac cagatacctg cggatccacc ctcagagttg ggtgcaccag     4320

attgctctgc ggatggaagt gctgggatgc gaggcccagg acctgtactg ataa           4374


<210>  4
<211>  5045
<212>  DNA
<213>  Artificial

<220>
<223>  Recombinant AAV vector sequence

<400>  4
ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc       60

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg      120

gccaactcca tcactagggg ttcctagatc tacgcgtctg tttactctgg ttaattttta      180

aaggagggta aacagtgcct gaaagctgac ctttgcccac attcctccgg tagacattaa      240

cttattaaat tgattctgat tacaaatctg acctttgccc ccatctcacc cagtaacaat      300

gcaagagttg atgtcagtct ataaaaagcg aagcgcgcgg tgggcggggt tcgctgcctg      360

caggtgagta tctcagggat ccagacatgg ggatatggga ggtgcctctg atcccagggc      420

tcactgtggg tctctctgtt cacagcctgc tagcgccacc atgcagatcg agctgtctac      480

ctgcttcttc ctgtgcctgc tgcggttctg cttcagcgcc accagacggt actatctggg      540

cgccgtggaa ctgagctggg actacatgca gagcgacctg ggcgagctgc ccgtggatgc      600

cagattccct ccaagagtgc ccaagagctt ccccttcaac acctccgtgg tgtacaagaa      660

aaccctgttc gtggaattca ccgaccacct gttcaatatc gccaagccca gacccccctg      720

gatgggcctg ctgggaccta caattcaggc cgaggtgtac gacaccgtcg tgatcaccct      780

gaagaacatg gccagccacc ccgtgtctct gcatgccgtg ggagtgtcct actggaaggc      840

ctctgagggc gccgagtacg acgatcagac cagccagcgc gagaaagagg acgacaaggt      900

gttccctggc ggcagccaca cctacgtgtg gcaggtgctg aaagaaaacg gccccatggc      960

ctccgaccct ctgtgcctga catacagcta cctgagccac gtggacctcg tgaaggacct     1020

gaacagcggc ctgatcggag ccctgctcgt gtgtagagag ggcagcctgg ccaaagagaa     1080

aacccagacc ctgcacaagt tcatcctgct gttcgccgtg ttcgacgagg gcaagagctg     1140

gcacagcgag acaaagaaca gcctgatgca ggaccgggac gccgcctctg ctagagcctg     1200

gcccaaaatg cacaccgtga acggctacgt gaacagaagc ctgcccggac tgatcggctg     1260

ccaccggaag tctgtgtact ggcacgtgat cggcatgggc accacccctg aggtgcacag     1320

catctttctg gaaggacaca cctttctcgt gcggaaccac cggcaggcca gcctggaaat     1380

cagccctatc accttcctga ccgcccagac actgctgatg gacctgggcc agtttctgct     1440

gttctgccac atcagctccc accagcacga cggcatggaa gcctacgtga aggtggacag     1500

ctgccccgag gaaccccagc tgcggatgaa gaacaacgag gaagccgagg actacgacga     1560

cgacctgacc gacagcgaga tggacgtggt gcgcttcgac gacgataaca gccccagctt     1620

catccagatc agaagcgtgg ccaagaagca ccccaagacc tgggtgcact atatcgccgc     1680

cgaggaagag gactgggatt acgcccctct ggtgctggcc cccgacgaca gaagctacaa     1740

gagccagtac ctgaacaatg gcccccagcg gatcggccgg aagtataaga aagtgcggtt     1800

catggcctac accgacgaga cattcaagac cagagaggcc atccagcacg agagcggcat     1860

cctgggccct ctgctgtatg gcgaagtggg cgacaccctg ctgatcatct tcaagaacca     1920

ggccagcaga ccctacaaca tctaccctca cggcatcacc gacgtgcggc ccctgtactc     1980

tagaaggctg cccaagggcg tgaaacacct gaaggacttc cccatcctgc ccggcgagat     2040

tttcaagtac aagtggaccg tgaccgtgga agatggcccc accaagagcg accccagatg     2100

cctgacacgg tactacagca gcttcgtgaa catggaacgg gacctggcct ccggcctgat     2160

tggcccactg ctgatctgct acaaagaaag cgtggaccag cggggcaacc agatcatgag     2220

cgacaagcgg aacgtgatcc tgtttagcgt gttcgatgag aaccggtcct ggtatctgac     2280

cgagaatatc cagcggttcc tgcccaaccc tgccggcgtg cagctggaag atcctgagtt     2340

ccaggcctcc aacatcatgc actccatcaa tggctatgtg ttcgacagcc tgcagctgag     2400

cgtgtgcctg cacgaggtgg cctactggta catcctgagc atcggggccc agaccgactt     2460

cctgtccgtg ttcttctccg gctacacctt caagcacaag atggtgtacg aggataccct     2520

gaccctgttc ccctttagcg gcgaaaccgt gttcatgagc atggaaaacc ccggcctgtg     2580

gatcctgggc tgccacaaca gcgacttccg gaacagaggc atgaccgccc tgctgaaggt     2640

gtccagctgc gacaagaaca ccggcgacta ctacgaggac agctatgagg acatcagcgc     2700

ctacctgctg agcaagaaca atgccatcga gcccagaagc ttcagccagc cccctgtgct     2760

gaagcggcac cagagagaga tcacccggac caccctgcag tccgaccagg aagagatcga     2820

ttacgacgac accatcagcg tggaaatgaa gaaagaagat ttcgacatct acgacgagga     2880

cgagaaccag agcccccggt cctttcagaa aaagacccgg cactacttca ttgccgctgt     2940

ggaacggctg tgggactacg gcatgagcag cagccctcac gtgctgagaa acagggccca     3000

gagcggcagc gtgccccagt tcaagaaagt ggtgttccag gaattcacag acggcagctt     3060

tacccagcct ctgtaccgcg gcgagctgaa cgaacacctg ggactgctgg gcccctatat     3120

ccgggccgaa gtggaagata acatcatggt caccttccgg aatcaggcct cccggcccta     3180

cagcttctac agctccctga tcagctacga agaggaccag agacagggcg ctgagccccg     3240

gaagaacttc gtgaagccca acgagactaa gacctacttt tggaaggtgc agcaccacat     3300

ggcccctaca aaggacgagt tcgactgcaa ggcctgggcc tacttctccg atgtggacct     3360

ggaaaaggac gtgcactctg ggctgatcgg ccccctgctc gtgtgccaca ccaacaccct     3420

gaatcccgcc cacggcagac aagtgacagt gcaggaattc gccctgttct tcaccatctt     3480

cgacgaaaca aagagctggt acttcaccga aaacatggaa agaaactgcc gggctccctg     3540

caacatccag atggaagatc ccaccttcaa agagaactac cggttccacg ccatcaacgg     3600

ctacatcatg gacacactgc ccggcctcgt gatggctcag gatcagcgga tccggtggta     3660

tctgctgtcc atgggctcca acgagaacat ccacagcatc cacttcagcg gccacgtgtt     3720

caccgtgcgg aaaaaagaag agtacaaaat ggccctgtac aacctgtacc ctggggtgtt     3780

cgagacagtg gaaatgctgc ccagcaaggc cggcatctgg cgggtggaat gtctgatcgg     3840

cgagcatctg cacgctggga tgagcacact gtttctggtg tacagcaaca agtgccagac     3900

acctctgggc atggcctctg gccacatccg ggactttcag atcacagcca gcggccagta     3960

tggccagtgg gccccaaaac tggccagact gcactacagc ggcagcatca acgcctggtc     4020

caccaaagag cccttcagct ggatcaaggt ggacctgctg gctcccatga tcatccacgg     4080

aatcaagacc cagggcgcca gacagaagtt ctccagcctg tacatctccc agttcatcat     4140

catgtactcc ctggacggca agaagtggca gacctaccgg ggcaatagca ccggcaccct     4200

gatggtgttc ttcggcaacg tggactccag cggcattaag cacaacatct tcaacccccc     4260

catcattgcc cggtacatcc ggctgcaccc cacccactac agcatccggt ccaccctgag     4320

aatggaactg atgggctgcg acctgaactc ctgcagcatg cccctgggga tggaaagcaa     4380

ggccatctcc gacgcccaga tcaccgcctc cagctacttc accaacatgt tcgccacctg     4440

gtccccatcc aaggcccggc tgcatctgca gggcagaagc aatgcttgga ggccccaagt     4500

gaacaacccc aaagaatggc tgcaggtgga cttccagaaa accatgaaag tgaccggcgt     4560

gaccacccag ggcgtgaagt ctctgctgac ctctatgtac gtgaaagagt tcctgatctc     4620

cagcagccag gacggccacc agtggaccct gtttttccag aacggcaaag tgaaagtgtt     4680

tcaggggaac caggacagct tcacccccgt cgtgaatagc ctggaccctc cactgctgac     4740

cagatacctg cggatccacc ctcagagttg ggtgcaccag attgctctgc ggatggaagt     4800

gctgggatgc gaggcccagg acctgtactg ataagtcgac aggcctaata aagagctcag     4860

atgcatcgat cagagtgtgt tggttttttg tgtgagatct aggaacccct agtgatggag     4920

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg     4980

cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg     5040

gccaa                                                                 5045


<210>  5
<211>  2333
<212>  PRT
<213>  Homo sapiens

<400>  5

Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr 
1               5                   10                  15      


Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro 
            20                  25                  30          


Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys 
        35                  40                  45              


Thr Leu Phe Val Glu Phe Thr Val His Leu Phe Asn Ile Ala Lys Pro 
    50                  55                  60                  


Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val 
65                  70                  75                  80  


Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val 
                85                  90                  95      


Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala 
            100                 105                 110         


Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val 
        115                 120                 125             


Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn 
    130                 135                 140                 


Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser 
145                 150                 155                 160 


His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu 
                165                 170                 175     


Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu 
            180                 185                 190         


His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp 
        195                 200                 205             


His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser 
    210                 215                 220                 


Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg 
225                 230                 235                 240 


Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His 
                245                 250                 255     


Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu 
            260                 265                 270         


Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile 
        275                 280                 285             


Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly 
    290                 295                 300                 


Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met 
305                 310                 315                 320 


Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg 
                325                 330                 335     


Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp 
            340                 345                 350         


Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe 
        355                 360                 365             


Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His 
    370                 375                 380                 


Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu 
385                 390                 395                 400 


Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro 
                405                 410                 415     


Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr 
            420                 425                 430         


Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile 
        435                 440                 445             


Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile 
    450                 455                 460                 


Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile 
465                 470                 475                 480 


Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys 
                485                 490                 495     


His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys 
            500                 505                 510         


Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys 
        515                 520                 525             


Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala 
    530                 535                 540                 


Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp 
545                 550                 555                 560 


Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe 
                565                 570                 575     


Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln 
            580                 585                 590         


Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe 
        595                 600                 605             


Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser 
    610                 615                 620                 


Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu 
625                 630                 635                 640 


Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr 
                645                 650                 655     


Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro 
            660                 665                 670         


Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp 
        675                 680                 685             


Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala 
    690                 695                 700                 


Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu 
705                 710                 715                 720 


Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala 
                725                 730                 735     


Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg 
            740                 745                 750         


Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys 
        755                 760                 765             


Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Ile Gln Asn 
    770                 775                 780                 


Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro 
785                 790                 795                 800 


His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe 
                805                 810                 815     


Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser 
            820                 825                 830         


Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val 
        835                 840                 845             


Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly 
    850                 855                 860                 


Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser 
865                 870                 875                 880 


Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala 
                885                 890                 895     


Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His 
            900                 905                 910         


Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro 
        915                 920                 925             


Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Glu Glu Asn Asn Asp 
    930                 935                 940                 


Ser Lys Leu Leu Glu Ser Gly Leu Met Asn Ser Gln Glu Ser Ser Trp 
945                 950                 955                 960 


Gly Lys Asn Val Ser Ser Thr Glu Ser Gly Arg Leu Phe Lys Gly Lys 
                965                 970                 975     


Arg Ala His Gly Pro Ala Leu Leu Thr Lys Asp Asn Ala Leu Phe Lys 
            980                 985                 990         


Val Ser Ile Ser Leu Leu Lys Thr  Asn Lys Thr Ser Asn  Asn Ser Ala 
        995                 1000                 1005             


Thr Asn  Arg Lys Thr His Ile  Asp Gly Pro Ser Leu  Leu Ile Glu 
    1010                 1015                 1020             


Asn Ser  Pro Ser Val Trp Gln  Asn Ile Leu Glu Ser  Asp Thr Glu 
    1025                 1030                 1035             


Phe Lys  Lys Val Thr Pro Leu  Ile His Asp Arg Met  Leu Met Asp 
    1040                 1045                 1050             


Lys Asn  Ala Thr Ala Leu Arg  Leu Asn His Met Ser  Asn Lys Thr 
    1055                 1060                 1065             


Thr Ser  Ser Lys Asn Met Glu  Met Val Gln Gln Lys  Lys Glu Gly 
    1070                 1075                 1080             


Pro Ile  Pro Pro Asp Ala Gln  Asn Pro Asp Met Ser  Phe Phe Lys 
    1085                 1090                 1095             


Met Leu  Phe Leu Pro Glu Ser  Ala Arg Trp Ile Gln  Arg Thr His 
    1100                 1105                 1110             


Gly Lys  Asn Ser Leu Asn Ser  Gly Gln Gly Pro Ser  Pro Lys Gln 
    1115                 1120                 1125             


Leu Val  Ser Leu Gly Pro Glu  Lys Ser Val Glu Gly  Gln Asn Phe 
    1130                 1135                 1140             


Leu Ser  Glu Lys Asn Lys Val  Val Val Gly Lys Gly  Glu Phe Thr 
    1145                 1150                 1155             


Lys Asp  Val Gly Leu Lys Glu  Met Val Phe Pro Ser  Ser Arg Asn 
    1160                 1165                 1170             


Leu Phe  Leu Thr Asn Leu Asp  Asn Leu His Glu Asn  Asn Thr His 
    1175                 1180                 1185             


Asn Gln  Glu Lys Lys Ile Gln  Glu Glu Ile Glu Lys  Lys Glu Thr 
    1190                 1195                 1200             


Leu Ile  Gln Glu Asn Val Val  Leu Pro Gln Ile His  Thr Val Thr 
    1205                 1210                 1215             


Gly Thr  Lys Asn Phe Met Lys  Asn Leu Phe Leu Leu  Ser Thr Arg 
    1220                 1225                 1230             


Gln Asn  Val Glu Gly Ser Tyr  Glu Gly Ala Tyr Ala  Pro Val Leu 
    1235                 1240                 1245             


Gln Asp  Phe Arg Ser Leu Asn  Asp Ser Thr Asn Arg  Thr Lys Lys 
    1250                 1255                 1260             


His Thr  Ala His Phe Ser Lys  Lys Gly Glu Glu Glu  Asn Leu Glu 
    1265                 1270                 1275             


Gly Leu  Gly Asn Gln Thr Lys  Gln Ile Val Glu Lys  Tyr Ala Cys 
    1280                 1285                 1290             


Thr Thr  Arg Ile Ser Pro Asn  Thr Ser Gln Gln Asn  Phe Val Thr 
    1295                 1300                 1305             


Gln Arg  Ser Lys Arg Ala Leu  Lys Gln Phe Arg Leu  Pro Leu Glu 
    1310                 1315                 1320             


Glu Thr  Glu Leu Glu Lys Arg  Ile Ile Val Asp Asp  Thr Ser Thr 
    1325                 1330                 1335             


Gln Trp  Ser Lys Asn Met Lys  His Leu Thr Pro Ser  Thr Leu Thr 
    1340                 1345                 1350             


Gln Ile  Asp Tyr Asn Glu Lys  Glu Lys Gly Ala Ile  Thr Gln Ser 
    1355                 1360                 1365             


Pro Leu  Ser Asp Cys Leu Thr  Arg Ser His Ser Ile  Pro Gln Ala 
    1370                 1375                 1380             


Asn Arg  Ser Pro Leu Pro Ile  Ala Lys Val Ser Ser  Phe Pro Ser 
    1385                 1390                 1395             


Ile Arg  Pro Ile Tyr Leu Thr  Arg Val Leu Phe Gln  Asp Asn Ser 
    1400                 1405                 1410             


Ser His  Leu Pro Ala Ala Ser  Tyr Arg Lys Lys Asp  Ser Gly Val 
    1415                 1420                 1425             


Gln Glu  Ser Ser His Phe Leu  Gln Gly Ala Lys Lys  Asn Asn Leu 
    1430                 1435                 1440             


Ser Leu  Ala Ile Leu Thr Leu  Glu Met Thr Gly Asp  Gln Arg Glu 
    1445                 1450                 1455             


Val Gly  Ser Leu Gly Thr Ser  Ala Thr Asn Ser Val  Thr Tyr Lys 
    1460                 1465                 1470             


Lys Val  Glu Asn Thr Val Leu  Pro Lys Pro Asp Leu  Pro Lys Thr 
    1475                 1480                 1485             


Ser Gly  Lys Val Glu Leu Leu  Pro Lys Val His Ile  Tyr Gln Lys 
    1490                 1495                 1500             


Asp Leu  Phe Pro Thr Glu Thr  Ser Asn Gly Ser Pro  Gly His Leu 
    1505                 1510                 1515             


Asp Leu  Val Glu Gly Ser Leu  Leu Gln Gly Thr Glu  Gly Ala Ile 
    1520                 1525                 1530             


Lys Trp  Asn Glu Ala Asn Arg  Pro Gly Lys Val Pro  Phe Leu Arg 
    1535                 1540                 1545             


Val Ala  Thr Glu Ser Ser Ala  Lys Thr Pro Ser Lys  Leu Leu Asp 
    1550                 1555                 1560             


Pro Leu  Ala Trp Asp Asn His  Tyr Gly Thr Gln Ile  Pro Lys Glu 
    1565                 1570                 1575             


Glu Trp  Lys Ser Gln Glu Lys  Ser Pro Glu Lys Thr  Ala Phe Lys 
    1580                 1585                 1590             


Lys Lys  Asp Thr Ile Leu Ser  Leu Asn Ala Cys Glu  Ser Asn His 
    1595                 1600                 1605             


Ala Ile  Ala Ala Ile Asn Glu  Gly Gln Asn Lys Pro  Glu Ile Glu 
    1610                 1615                 1620             


Val Thr  Trp Ala Lys Gln Gly  Arg Thr Glu Arg Leu  Cys Ser Gln 
    1625                 1630                 1635             


Asn Pro  Pro Val Leu Lys Arg  His Gln Arg Glu Ile  Thr Arg Thr 
    1640                 1645                 1650             


Thr Leu  Gln Ser Asp Gln Glu  Glu Ile Asp Tyr Asp  Asp Thr Ile 
    1655                 1660                 1665             


Ser Val  Glu Met Lys Lys Glu  Asp Phe Asp Ile Tyr  Asp Glu Asp 
    1670                 1675                 1680             


Glu Asn  Gln Ser Pro Arg Ser  Phe Gln Lys Lys Thr  Arg His Tyr 
    1685                 1690                 1695             


Phe Ile  Ala Ala Val Glu Arg  Leu Trp Asp Tyr Gly  Met Ser Ser 
    1700                 1705                 1710             


Ser Pro  His Val Leu Arg Asn  Arg Ala Gln Ser Gly  Ser Val Pro 
    1715                 1720                 1725             


Gln Phe  Lys Lys Val Val Phe  Gln Glu Phe Thr Asp  Gly Ser Phe 
    1730                 1735                 1740             


Thr Gln  Pro Leu Tyr Arg Gly  Glu Leu Asn Glu His  Leu Gly Leu 
    1745                 1750                 1755             


Leu Gly  Pro Tyr Ile Arg Ala  Glu Val Glu Asp Asn  Ile Met Val 
    1760                 1765                 1770             


Thr Phe  Arg Asn Gln Ala Ser  Arg Pro Tyr Ser Phe  Tyr Ser Ser 
    1775                 1780                 1785             


Leu Ile  Ser Tyr Glu Glu Asp  Gln Arg Gln Gly Ala  Glu Pro Arg 
    1790                 1795                 1800             


Lys Asn  Phe Val Lys Pro Asn  Glu Thr Lys Thr Tyr  Phe Trp Lys 
    1805                 1810                 1815             


Val Gln  His His Met Ala Pro  Thr Lys Asp Glu Phe  Asx Asp Cys 
    1820                 1825                 1830             


Lys Ala  Trp Ala Tyr Phe Ser  Asp Val Asp Leu Glu  Lys Asp Val 
    1835                 1840                 1845             


His Ser  Gly Leu Ile Gly Pro  Leu Leu Val Cys His  Thr Asn Thr 
    1850                 1855                 1860             


Leu Asn  Pro Ala His Gly Arg  Gln Val Thr Val Gln  Glu Phe Ala 
    1865                 1870                 1875             


Leu Phe  Phe Thr Ile Phe Asp  Glu Thr Lys Ser Trp  Tyr Phe Thr 
    1880                 1885                 1890             


Glu Asn  Met Glu Arg Asn Cys  Arg Ala Pro Cys Asn  Ile Gln Met 
    1895                 1900                 1905             


Glu Asp  Pro Thr Phe Lys Glu  Asn Tyr Arg Phe His  Ala Ile Asn 
    1910                 1915                 1920             


Gly Tyr  Ile Met Asp Thr Leu  Pro Gly Leu Val Met  Ala Gln Asp 
    1925                 1930                 1935             


Gln Arg  Ile Arg Trp Tyr Leu  Leu Ser Met Gly Ser  Asn Glu Asn 
    1940                 1945                 1950             


Ile His  Ser Ile His Phe Ser  Gly His Val Phe Thr  Val Arg Lys 
    1955                 1960                 1965             


Lys Glu  Glu Tyr Lys Met Ala  Leu Tyr Asn Leu Tyr  Pro Gly Val 
    1970                 1975                 1980             


Phe Glu  Thr Val Glu Met Leu  Pro Ser Lys Ala Gly  Ile Trp Arg 
    1985                 1990                 1995             


Val Glu  Cys Leu Ile Gly Glu  His Leu His Ala Gly  Met Ser Thr 
    2000                 2005                 2010             


Leu Phe  Leu Val Tyr Ser Asn  Lys Cys Gln Thr Pro  Leu Gly Met 
    2015                 2020                 2025             


Ala Ser  Gly His Ile Arg Asp  Phe Gln Ile Thr Ala  Ser Gly Gln 
    2030                 2035                 2040             


Tyr Gly  Gln Trp Ala Pro Lys  Leu Ala Arg Leu His  Tyr Ser Gly 
    2045                 2050                 2055             


Ser Ile  Asn Ala Trp Ser Thr  Lys Glu Pro Phe Ser  Trp Ile Lys 
    2060                 2065                 2070             


Val Asp  Leu Leu Ala Pro Met  Ile Ile His Gly Ile  Lys Thr Gln 
    2075                 2080                 2085             


Gly Ala  Arg Gln Lys Phe Ser  Ser Leu Tyr Ile Ser  Gln Phe Ile 
    2090                 2095                 2100             


Ile Met  Tyr Ser Leu Asp Gly  Lys Lys Trp Gln Thr  Tyr Arg Gly 
    2105                 2110                 2115             


Asn Ser  Thr Gly Thr Leu Met  Val Phe Phe Gly Asn  Val Asp Ser 
    2120                 2125                 2130             


Ser Gly  Ile Lys His Asn Ile  Phe Asn Pro Pro Ile  Ile Ala Arg 
    2135                 2140                 2145             


Tyr Ile  Arg Leu His Pro Thr  His Tyr Ser Ile Arg  Ser Thr Leu 
    2150                 2155                 2160             


Arg Met  Glu Leu Met Gly Cys  Asp Leu Asn Ser Cys  Ser Met Pro 
    2165                 2170                 2175             


Leu Gly  Met Glu Ser Lys Ala  Ile Ser Asp Ala Gln  Ile Thr Ala 
    2180                 2185                 2190             


Ser Ser  Tyr Phe Thr Asn Met  Phe Ala Thr Trp Ser  Pro Ser Lys 
    2195                 2200                 2205             


Ala Arg  Leu His Leu Gln Gly  Arg Ser Asn Ala Trp  Arg Pro Gln 
    2210                 2215                 2220             


Val Asn  Asn Pro Lys Glu Trp  Leu Gln Val Asp Phe  Gln Lys Thr 
    2225                 2230                 2235             


Met Lys  Val Thr Gly Val Thr  Thr Gln Gly Val Lys  Ser Leu Leu 
    2240                 2245                 2250             


Thr Ser  Met Tyr Val Lys Glu  Phe Leu Ile Ser Ser  Ser Gln Asp 
    2255                 2260                 2265             


Gly His  Gln Trp Thr Leu Phe  Phe Gln Asn Gly Lys  Val Lys Val 
    2270                 2275                 2280             


Phe Gln  Gly Asn Gln Asp Ser  Phe Thr Pro Val Val  Asn Ser Leu 
    2285                 2290                 2295             


Asp Pro  Pro Leu Leu Thr Arg  Tyr Leu Arg Ile His  Pro Gln Ser 
    2300                 2305                 2310             


Trp Val  His Gln Ile Ala Leu  Arg Met Glu Val Leu  Gly Cys Glu 
    2315                 2320                 2325             


Ala Gln  Asp Leu Tyr 
    2330             


<210>  6
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII consensus insert sequence


<220>
<221>  MISC_FEATURE
<222>  (1)..(2)
<223>  Xaa can be S or T

<220>
<221>  MISC_FEATURE
<222>  (7)..(7)
<223>  Xaa can be S or T

<400>  6

Xaa Xaa Tyr Val Asn Arg Xaa Leu 
1               5               


<210>  7
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  7

Thr Thr Tyr Val Asn Arg Ser Leu 
1               5               


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

<220>
<223>  Factor VIII insert sequence

<400>  8

Thr Thr Tyr Val Asn Arg Thr Leu 
1               5               


<210>  9
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  9

Thr Ser Tyr Val Asn Arg Ser Leu 
1               5               


<210>  10
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  10

Thr Ser Tyr Val Asn Arg Thr Leu 
1               5               


<210>  11
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  11

Ser Thr Tyr Val Asn Arg Ser Leu 
1               5               


<210>  12
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  12

Ser Thr Tyr Val Asn Arg Thr Leu 
1               5               


<210>  13
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  13

Ser Ser Tyr Val Asn Arg Ser Leu 
1               5               


<210>  14
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  14

Ser Ser Tyr Val Asn Arg Thr Leu 
1               5               


<210>  15
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII consensus insert sequence


<220>
<221>  MISC_FEATURE
<222>  (1)..(2)
<223>  Xaa can be S or T

<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  Xaa can be S or T

<400>  15

Xaa Xaa Asn Asn Xaa 
1               5   


<210>  16
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  16

Thr Thr Asn Asn Ser 
1               5   


<210>  17
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  17

Thr Thr Asn Asn Thr 
1               5   


<210>  18
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  18

Thr Ser Asn Asn Ser 
1               5   


<210>  19
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  19

Thr Ser Asn Asn Thr 
1               5   


<210>  20
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  20

Ser Thr Asn Asn Ser 
1               5   


<210>  21
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  21

Ser Thr Asn Asn Thr 
1               5   


<210>  22
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  22

Ser Ser Asn Asn Ser 
1               5   


<210>  23
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Factor VIII insert sequence

<400>  23

Ser Ser Asn Asn Thr 
1               5   


<210>  24
<211>  7
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site consensus sequence


<220>
<221>  misc_feature
<222>  (2)..(3)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  MISC_FEATURE
<222>  (6)..(6)
<223>  Xaa can be S or T

<400>  24

Cys Xaa Xaa Gly Gly Xaa Cys 
1               5           


<210>  25
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site sequence


<220>
<221>  MISC_FEATURE
<222>  (4)..(4)
<223>  Xaa can be D or E

<400>  25

Asn Ser Thr Xaa Ala 
1               5   


<210>  26
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site sequence

<400>  26

Asn Ile Thr Gln Ser 
1               5   


<210>  27
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site sequence

<400>  27

Gln Ser Thr Gln Ser 
1               5   


<210>  28
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site sequence


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  Xaa can be D or E

<220>
<221>  MISC_FEATURE
<222>  (4)..(4)
<223>  Xaa can be R or K

<400>  28

Xaa Phe Thr Xaa Val 
1               5   


<210>  29
<211>  4
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site sequence


<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  Xaa can be D or E

<400>  29

Cys Xaa Ser Asn 
1               


<210>  30
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  O-linked glycosylation site sequence


<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  Xaa can be R or K

<400>  30

Gly Gly Ser Cys Xaa 
1               5   


<210>  31
<211>  30
<212>  PRT
<213>  Artificial

<220>
<223>  B domain deleted Factor VIII sequence

<400>  31

Leu Ser Lys Asn Asn Ala Ile Glu Pro Arg Ser Phe Ser Gln Asn Pro 
1               5                   10                  15      


Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr 
            20                  25                  30  


<210>  32
<211>  30
<212>  PRT
<213>  Artificial

<220>
<223>  Mutated B domain deleted Factor VIII sequence

<400>  32

Leu Ser Lys Asn Asn Thr Thr Glu Pro Arg Ser Phe Ser Gln Asn Pro 
1               5                   10                  15      


Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr 
            20                  25                  30  


<210>  33
<211>  31
<212>  PRT
<213>  Artificial

<220>
<223>  Mutated B domain deleted Factor VIII sequence

<400>  33

Leu Ser Lys Asn Asn Thr Thr Tyr Val Asn Arg Ser Leu Ser Gln Asn 
1               5                   10                  15      


Pro Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr 
            20                  25                  30      


<210>  34
<211>  28
<212>  PRT
<213>  Artificial

<220>
<223>  Mutated B domain deleted Factor VIII sequence

<400>  34

Leu Ser Lys Asn Asn Thr Thr Asn Asn Thr Ser Gln Asn Pro Pro Val 
1               5                   10                  15      


Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr 
            20                  25              


