                         SEQUENCE LISTING

<110>  Richter Gedeon Nyrt.
 
<120>  Novel cell-based TNF-alpha binding assay

<130>  GE46A19/P-WO

<150>  P1600456
<151>  2016-07-19

<160>  12    

<170>  PatentIn version 3.5

<210>  1
<211>  792
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of a TACE cleavage resistant version of membrane-bound 
       human TNF-alpha


<220>
<221>  misc_feature
<222>  (1)..(792)
<223>  Sequence of a TACE cleavage resistant version of membrane-bound 
       human TNF-alpha

<220>
<221>  misc_feature
<222>  (16)..(18)
<223>  start translation

<220>
<221>  mutation
<222>  (318)..(355)
<223>  mutagenic oligo

<220>
<221>  modified_base
<222>  (324)..(324)

<220>
<221>  modified_base
<222>  (329)..(330)

<220>
<221>  modified_base
<222>  (333)..(334)

<220>
<221>  misc_feature
<222>  (349)..(351)
<223>  TNF-alpha start

<220>
<221>  misc_feature
<222>  (784)..(786)
<223>  stop translation

<400>  1
gaattcgaca agcttatgat cgaaacatac aaccaaactt ctccccgatc tgcggccact       60

ggactgccca tcagcatgaa aatttttatg tatttactta ctgtttttct tatcacccag      120

atgattgggt cagcactttt tgctgtgtat cttcatagaa ggttggacaa gatagaagat      180

gaaaggaatc ttcatgaaga ttttgtattc atgaaaacga tacagagatg caacacagga      240

gaaagatcct tatccttact gaactgtgag gagattaaaa gccagtttga aggctttgtg      300

aaggatataa tgttaaacaa agaagagaaa aaagaagatg aggatcctgt agcccatgtt      360

gtagcaaacc ctcaagctga ggggcagctc cagtggctga accgccgggc caatgccctc      420

ctggccaatg gcgtggagct gagagataac cagctggtgg tgccatcaga gggcctgtac      480

ctcatctact cccaggtcct cttcaagggc caaggctgcc cctccaccca tgtgctcctc      540

acccacacca tcagccgcat cgccgtctcc taccagacca aggtcaacct cctctctgcc      600

atcaagagcc cctgccagag ggagacccca gagggggctg aggccaagcc ctggtatgag      660

cccatctatc tgggaggggt cttccagctg gagaagggtg accgactcag cgctgagatc      720

aatcggcccg actatctcga ctttgccgag tctgggcagg tctactttgg gatcattgct      780

ctgtgaatct ag                                                          792


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

<220>
<223>  Protease cleavage site wild type TNF-alpha


<220>
<221>  misc_feature
<222>  (1)..(21)
<223>  Protease cleavage site wild-type TNF-alpha

<400>  2
aaagaggaga cgaagaaaga t                                                 21


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

<220>
<223>  Protease cleavage site mutant TNF-alpha


<220>
<221>  misc_feature
<222>  (1)..(21)
<223>  Protease cleavage site mutant TNF-alpha

<220>
<221>  modified_base
<222>  (10)..(12)
<223>  amino acid  change (T -> K)

<220>
<221>  modified_base
<222>  (16)..(18)
<223>  amino acid  change (K -> E)

<400>  3
aaagaagaga aaaaagaaga t                                                 21


<210>  4
<211>  811
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Optimized transgene sequence for genomic integration


<220>
<221>  misc_feature
<222>  (1)..(811)
<223>  Optimized transgene sequence for genomic integration

<220>
<221>  misc_feature
<222>  (1)..(17)
<223>  5' and 3' flanking sequences

<220>
<221>  misc_feature
<222>  (351)..(353)
<223>  TNF-alpha sequence start

<220>
<221>  misc_feature
<222>  (789)..(811)
<223>  5' and 3' flanking sequences

<400>  4
ggatccgttt aaacgtcatg attgagacct acaaccagac ctcccctaga agtgctgcca       60

ccgggctgcc tattagtatg aagattttta tgtacctgct gacagtgttc ctgatcaccc      120

agatgattgg gagtgctctg tttgcagtct acctgcaccg gagactggac aagatcgagg      180

atgaaaggaa cctgcatgaa gacttcgtgt ttatgaaaac tattcagcga tgcaacaccg      240

gagagcggag cctgtccctg ctgaattgtg aggaaatcaa gtcccagttc gaagggtttg      300

tgaaggatat tatgctgaac aaagaggaaa agaaagagga cgaagatcct gtcgcccacg      360

tggtcgctaa tccacaggca gagggacagc tgcagtggct gaacaggcga gcaaatgctc      420

tgctggcaaa cggagtggaa ctgcgggaca atcagctggt ggtcccctcc gagggactgt      480

acctgatcta ttctcaggtg ctgttcaaag gacagggctg ccctagcaca cacgtcctgc      540

tgacccatac aatctcacgc attgccgtga gctatcagac aaaggtcaac ctgctgtctg      600

ccatcaagtc cccatgtcag cgagagactc ccgaaggagc agaggccaag ccctggtacg      660

agcctatcta tctgggcggg gtgtttcagc tggaaaaagg cgatagactg tctgccgaga      720

ttaataggcc cgactatctg gacttcgctg aatctgggca ggtgtatttt ggcattatcg      780

cactgtgatg actcgaggcg gccgcaagct t                                     811


<210>  5
<211>  7015
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pTriEx-4Neo-hrkmtnfa1 expression vector construct sequence with 
       insert


<220>
<221>  misc_feature
<222>  (1)..(7015)
<223>  pTriEx-4Neo-hrkmtnfa1 expression Vector construct sequence with 
       insert

<220>
<221>  misc_feature
<222>  (2051)..(2828)
<223>  synthetic membrane-bound TNF-alpha gene

<400>  5
tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga       60

ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat      120

gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca      180

tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa      240

ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt      300

attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa      360

cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag      420

gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt      480

ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt      540

ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt      600

aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg      660

catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc      720

tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt      780

gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt      840

aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc      900

tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg      960

cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggcct     1020

attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta     1080

cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt     1140

caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg     1200

cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga     1260

ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc     1320

tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc     1380

caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact     1440

ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt     1500

gggaggtcta tataagcaga cgtcgtttag tgaaccgtca gatcactaga tgctttattg     1560

cggtagttta tcacagttaa attgctaacg ccagtctcga acttaacgtg cagaagttgg     1620

tcgtgaggca ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg gctcggggct     1680

gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc     1740

gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac     1800

agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc     1860

ggaccgaaat taatacgact cactataggg gaattgtgag cggataacaa ttccccggag     1920

ttaatccggg acctttaatt caacccaaca caatatatta tagttaaata agaattatta     1980

tcaaatcatt tgtatattaa ttaaaatact atactgtaaa ttacatttta tttacaatca     2040

aaggagatat accatgattg agacctacaa ccagacctcc cctagaagtg ctgccaccgg     2100

gctgcctatt agtatgaaga tttttatgta cctgctgaca gtgttcctga tcacccagat     2160

gattgggagt gctctgtttg cagtctacct gcaccggaga ctggacaaga tcgaggatga     2220

aaggaacctg catgaagact tcgtgtttat gaaaactatt cagcgatgca acaccggaga     2280

gcggagcctg tccctgctga attgtgagga aatcaagtcc cagttcgaag ggtttgtgaa     2340

ggatattatg ctgaacaaag aggaaaagaa agaggacgaa gatcctgtcg cccacgtggt     2400

cgctaatcca caggcagagg gacagctgca gtggctgaac aggcgagcaa atgctctgct     2460

ggcaaacgga gtggaactgc gggacaatca gctggtggtc ccctccgagg gactgtacct     2520

gatctattct caggtgctgt tcaaaggaca gggctgccct agcacacacg tcctgctgac     2580

ccatacaatc tcacgcattg ccgtgagcta tcagacaaag gtcaacctgc tgtctgccat     2640

caagtcccca tgtcagcgag agactcccga aggagcagag gccaagccct ggtacgagcc     2700

tatctatctg ggcggggtgt ttcagctgga aaaaggcgat agactgtctg ccgagattaa     2760

taggcccgac tatctggact tcgctgaatc tgggcaggtg tattttggca ttatcgcact     2820

gtgatgactc gagcaccacc atcaccatca ccatcactaa gtgattaacc tcaggtgcag     2880

gctgcctatc agaaggtggt ggctggtgtg gtaattccgg ttattttcca ccatattgcc     2940

gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag     3000

gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt     3060

tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa     3120

ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc     3180

aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg     3240

gctcacctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat     3300

gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa     3360

acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc     3420

ttgccacaac ccgcgataat tcctgcagcc aatatgggat cggccattga acaagatgga     3480

ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa     3540

cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt     3600

ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg     3660

ctatcgtggc tggccacgac gggcgttcct tgcgcagcag tgctcgacgt tgtcactgaa     3720

gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac     3780

cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt     3840

gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact     3900

cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg     3960

ccagccgaac tgttcgccag gctcaaggcg cgtatgcccg acggcgatga tctcgtcgtg     4020

actcatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc     4080

atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt     4140

gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc     4200

gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaggg     4260

gatcaattct ctagccaatg ccctggctca caaataccac tgagatcgat ctttttccct     4320

ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa     4380

ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga     4440

catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa     4500

catatgccca tatgtaacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt     4560

ttttgctgaa agcatgcgga ggaaattctc cttgaagttt ccctggtgtt caaagtaaag     4620

gagtttgcac cagacgcacc tctgttcact ggtccggcgt attaaaacac gatacattgt     4680

tattagtaca tttattaagc gctagattct gtgcgttgtt gatttacaga caattgttgt     4740

acgtatttta ataattcatt aaatttataa tctttagggt ggtatgttag agcgaaaatc     4800

aaatgatttt cagcgtcttt atatctgaat ttaaatatta aatcctcaat agatttgtaa     4860

aataggtttc gattagtttc aaacaagggt tgtttttccg aaccgatggc tggactatct     4920

aatggatttt cgctcaacgc cacaaaactt gccaaatctt gtagcagcaa tctagctttg     4980

tcgatattcg tttgtgtttt gttttgtaat aaaggttcga cgtcgttcaa aatattatgc     5040

gcttttgtat ttctttcatc actgtcgtta gtgtacaatt gactcgacgt aaacacgtta     5100

aatagagctt ggacatattt aacatcgggc gtgttagctt tattaggccg attatcgtcg     5160

tcgtcccaac cctcgtcgtt agaagttgct tccgaagacg attttgccat agccacacga     5220

cgcctattaa ttgtgtcggc taacacgtcc gcgatcaaat ttgtagttga gctttttgga     5280

attgcgatcg cataacttcg tatagcatac attatacgaa gttataagct cggaacgctg     5340

cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta     5400

tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc     5460

aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc cctgacgagc     5520

atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc     5580

aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg     5640

gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta     5700

ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg     5760

ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac     5820

acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag     5880

gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat     5940

ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat     6000

ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc     6060

gcagaaaaaa aggatctcaa gaagatcctt tgttaccaat gcttaatcag tgaggcacct     6120

atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata     6180

actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca     6240

cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga     6300

agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga     6360

gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg     6420

gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga     6480

gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt     6540

gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct     6600

cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca     6660

ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat     6720

accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga     6780

aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc     6840

aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg     6900

caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc     6960

ctttttcaat attattgaag catttatcag ggttattgtc tcatgtccgc gcgtt          7015


<210>  6
<211>  13802
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pIM.RAG1.EF1al.Neo.2_hrtnfakm1 expression vector construct 
       sequence with insert


<220>
<221>  misc_feature
<222>  (1)..(13802)
<223>  pIM.RAG1.EF1al.Neo.2_hrtnfakm1 expression vector construct 
       sequence with insert

<400>  6
agagtttaaa cgtcatgatt gagacctaca accagacctc ccctagaagt gctgccaccg       60

ggctgcctat tagtatgaag atttttatgt acctgctgac agtgttcctg atcacccaga      120

tgattgggag tgctctgttt gcagtctacc tgcaccggag actggacaag atcgaggatg      180

aaaggaacct gcatgaagac ttcgtgttta tgaaaactat tcagcgatgc aacaccggag      240

agcggagcct gtccctgctg aattgtgagg aaatcaagtc ccagttcgaa gggtttgtga      300

aggatattat gctgaacaaa gaggaaaaga aagaggacga agatcctgtc gcccacgtgg      360

tcgctaatcc acaggcagag ggacagctgc agtggctgaa caggcgagca aatgctctgc      420

tggcaaacgg agtggaactg cgggacaatc agctggtggt cccctccgag ggactgtacc      480

tgatctattc tcaggtgctg ttcaaaggac agggctgccc tagcacacac gtcctgctga      540

cccatacaat ctcacgcatt gccgtgagct atcagacaaa ggtcaacctg ctgtctgcca      600

tcaagtcccc atgtcagcga gagactcccg aaggagcaga ggccaagccc tggtacgagc      660

ctatctatct gggcggggtg tttcagctgg aaaaaggcga tagactgtct gccgagatta      720

ataggcccga ctatctggac ttcgctgaat ctgggcaggt gtattttggc attatcgcac      780

tgtgatgact cgaggcggcc gcggagctcc aggaattctg cagatcactg tgccttctag      840

ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac      900

tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca      960

ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag     1020

caggcatgct ggggatgcgg tgggctctat ggatccgcag cctctttccc acccaccttg     1080

ggactcagtt ctgccccaga tgaaattcag cacccacata ttaaattttc agaatggaaa     1140

tttaagctgt tccgggtgag atcctttgaa aagacacctg aagaagctca aaaggaaaag     1200

aaggattcct ttgaggggaa accctctctg gagcaatctc cagcagtcct ggacaaggct     1260

gatggtcaga agccagtccc aactcagcca ttgttaaaag cccaccctaa gttttcgaag     1320

aaatttcacg acaacgagaa agcaagaggc aaagcgatcc atcaagccaa ccttcgacat     1380

ctctgccgca tctgtgggaa ttcttttaga gctgatgagc acaacaggag atatccagtc     1440

catggtcctg tggatggtaa aaccctaggc cttttacgaa agaaggaaaa gagagctact     1500

tcctggccgg acctcattgc caaggttttc cggatcgatg tgaaggcaga tgttgactcg     1560

atccacccca ctgagttctg ccataactgc tggagcatca tgcacaggaa gtttagcagt     1620

gccccatgtg aggtttactt cccgaggaac gtgaccatgg agtggcaccc ccacacacca     1680

tcctgtgaca tctgcaacac tgcccgtcgg ggactcaaga ggaagagtct tcagccaaac     1740

ttgcagctca gcaaaaaact caaaactgtg cttgaccaag caagacaagc ccgtcagcac     1800

aagagaagag ctcaggcaag gatcagcagc aaggatgtca tgaagaagat cgccaactgc     1860

agtaagatac atcttagtac caagctcctt gcagtggact tcccagagca ctttgtgaaa     1920

tccatctcct gccagatctg tgaacacatt ctggctgacc ctgtggagac caactgtaag     1980

catgtctttt gccgggtctg cattctcaga tgcctcaaag tcatgggcag ctattgtccc     2040

tcttgccgat atccatgctt ccctactgac ctggagagtc cagtgaagtc ctttctgagc     2100

gtcttgaatt ccctgatggt gaaatgtcca gcaaaagagt gcaatgagga ggtcagtttg     2160

gaaaaatata atcaccacat ctcaagtcac aaggaatcaa aagagatttt tgtgcacatt     2220

aataaagggg gtcgagtaac gcgtgcaggc atgcaagctg gccgcaataa aatatcttta     2280

ttttcattac atctgtgtgt tggttttttg tgtgaatcgt aactaacata cgctctccat     2340

caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt     2400

gccagaacat ttctctatcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag     2460

cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc     2520

tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt     2580

cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc     2640

aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc     2700

gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc     2760

gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga     2820

ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt     2880

tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac     2940

agatccaagc tgtgaccggc gcctacgtaa gtgatatcta ctagatttat caaaaagagt     3000

gttgacttgt gagcgctcac aattgatact tagattcatc gagagggaca cgtcgactac     3060

taaccttctt ctctttccta cagctgagat caccggcgaa ggagggccac catggcttct     3120

taccctggac accagcatgc ttctgccttt gaccaggctg ccagatccag gggccactcc     3180

aacaggagaa ctgccctaag acccagaaga cagcaggaag ccactgaggt gaggcctgag     3240

cagaagatgc caaccctgct gagggtgtac attgatggac ctcatggcat gggcaagacc     3300

accaccactc aactgctggt ggcactgggc tccagggatg acattgtgta tgtgcctgag     3360

ccaatgacct actggagagt gctaggagcc tctgagacca ttgccaacat ctacaccacc     3420

cagcacaggc tggaccaggg agaaatctct gctggagatg ctgctgtggt gatgacctct     3480

gcccagatca caatgggaat gccctatgct gtgactgatg ctgttctggc tcctcacatt     3540

ggaggagagg ctggctcttc tcatgcccct ccacctgccc tgaccctgat ctttgacaga     3600

caccccattg cagccctgct gtgctaccca gcagcaaggt acctcatggg ctccatgacc     3660

ccacaggctg tgctggcttt tgtggccctg atccctccaa ccctccctgg caccaacatt     3720

gttctgggag cactgcctga agacagacac attgacaggc tggcaaagag gcagagacct     3780

ggagagagac tggacctggc catgctggct gcaatcagaa gggtgtatgg actgctggca     3840

aacactgtga gatacctcca gtgtggaggc tcttggagag aggactgggg acagctctct     3900

ggaacagcag tgccccctca aggagctgag ccccagtcca atgctggtcc aagaccccac     3960

attggggaca ccctgttcac cctgttcaga gcccctgagc tgctggctcc caatggagac     4020

ctgtacaatg tgtttgcctg ggctctggat gttctagcca agaggctgag gtccatgcat     4080

gtgttcatcc tggactatga ccagtcccct gctggatgca gagatgctct gctgcaacta     4140

acctctggca tggtgcagac ccatgtgacc acccctggca gcatccccac catctgtgac     4200

ctagccagaa cctttgccag ggagatggga gaggccaact aaacctgagc tagctcgaca     4260

tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct     4320

ttatttgtga aatttgtgat gctattgctt tatttgtgaa atttgtgatg ctattgcttt     4380

atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat     4440

gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg     4500

tggtagatcc atttttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc     4560

cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct     4620

aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa     4680

acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta     4740

ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc     4800

gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg     4860

caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt     4920

tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa     4980

gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct     5040

ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc     5100

cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg     5160

tcgttcgctc caagctgggc tgtgtgcacg acccccccgt tcagcccgac cgctgcgcct     5220

tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag     5280

cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga     5340

agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga     5400

agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg     5460

gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag     5520

aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag     5580

ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat     5640

gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct     5700

taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac     5760

tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa     5820

tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg     5880

gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt     5940

gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca     6000

ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt     6060

cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct     6120

tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg     6180

cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg     6240

agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg     6300

cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa     6360

aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt     6420

aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt     6480

gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt     6540

gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca     6600

tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat     6660

ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata     6720

aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc     6780

tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca     6840

gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg     6900

cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat     6960

gcgtaaggag aaaataccgc atcaggcgcc aatattaaac ttgatgagct ctagagatgg     7020

tcatgcattt taaaaagaat tactcaaaat attgtcttgg aataccagag agcaagtgct     7080

ttaagtatag gctgggaagt aaaatgctaa aggaatgaga aggcatttgg ggttgagttc     7140

aacctaagag gcaggggagc cacagggaaa gacctagcac ctgccacaga agagaattag     7200

gaagcagaat tgaactataa gcaattttga ggtgttcgtt gggctgcagt tgaaatattt     7260

tttgaggtta atgagacatt tgaaatggcc gtgtattgtt taactcttgc atagtcctgc     7320

atagggaaca atctaatagg atttctctgt gaatcaagtc ttagaaattt gcttttaatt     7380

tttatgaaaa acgcccattt ctttgttttt gagacagagt cctgctctgt catccaggct     7440

gggttgcagt ggcgtgatct tggcccactg caatctctgc ctcctgggtt caggcaattt     7500

tcctgtctca gcctcccgag tagctgggat ttcaagtgcc tgccaccatg cccggctaaa     7560

tttttttgta tttttggtac agatggagta tcaccatgtt ggccaggctg gtctcgaact     7620

cctgacctca agtgattcac cagccttgac ctcccaaagt gttgggatca caggcatgag     7680

ccactgtgcc tgtgccccaa aacaccaatt tctgatgtgt gatgcatgta agatagaaca     7740

aacttcagta aagcggggac ttgaaaagag gctttggtaa cagctgtcag cattaaccct     7800

tgcccctccg tacctcctaa tcccacccct gctcaaagta tgttcatctg agaatttgtc     7860

tccataacta tgtgactata aaaattctca tcgattttgt tagttgatca attgagggaa     7920

aaacatatgt tacttgatat aactggtggg tcaaaagaat taacccaggc aaatttgaga     7980

taggtggatg ggatgatgga ttgaaaatac agctgctctc tttccaatca tgtactaagt     8040

aatttgggaa agattgatct aattgggtct agagagtaca cttcacatgg cattgtttga     8100

ctttttttct gcatcgctag cgatctgtgc attacaactc aaatcagtcg ggtttcctgg     8160

catatgtaat tgccaatgtt ttttaccaga agagaaacat tactcccacc tcttcttatt     8220

atgttacaaa ctatagtgct aatgaccatc gaccaacagt gactttcagg atgacctgtg     8280

tgagttttat ctgaaaccat gtgaattttt catcttaaaa gtcccttaga atctcagtct     8340

atgtacactc aggtttgttg caggtttaga gttccgtgtt ttttgtttct aatgtagaca     8400

cagccttata atttacaaca gcattcacta attaaaattg taagcataat tactatccac     8460

gatacttatt attagtttgc attcataaag ctcaaaattc acttcatcct ttcaagtagt     8520

gaataattag tttctttggg tttgcagctt tatcatcctt ttatgaccca tttggaagaa     8580

ataaacaacc aaccccctgg aagactgctt taaaaagctg gaaatacatt gtccagctag     8640

tacaatgagg ctaatacaat gtggaaaata ttacttttct ttgattttag tagcctgttt     8700

atctttacat ttactgaaca aataactatt gagcacctaa tgtatactgg gacgacggta     8760

tcgataaggg cgcgccaaag ctaactgtag gactgagtct attctaaact gaaagcctgg     8820

acatctggag taccaggggg agatgacgtg ttacgggctt ccataaaagc agctggcttt     8880

gaatggaagg agccaagagg ccagcacagg agcggattcg tcgctttcac ggccatcgag     8940

ccgaacctct cgcaagtccg tgagccgtta aggaggcccc cagtcccgac ccttcgcccc     9000

aagcccctcg gggtccccgg gcctggtact ccttgccaca cgggaggggc gcggaagccg     9060

gggcggagga ggagccaacc ccgggctggg ctgagacccg cagaggaaga cgctctaggg     9120

atttgtcccg gactagcgag atggcaaggc tgaggacggg aggctgattg agaggcgaag     9180

gtacacccta atctcaatac aacctttgga gctaagccag caatggtaga gggaagattc     9240

tgcacgtccc ttccaggcgg cctccccgtc accacccccc ccaacccgcc ccgaccggag     9300

ctgagagtaa ttcatacaaa aggactcgcc cctgccttgg ggaatcccag ggaccgtcgt     9360

taaactccca ctaacgtaga acccagagat cgctgcgttc ccgccccctc acccgcccgc     9420

tctcgtcatc actgaggtgg agaagagcat gcgtgaggct ccggtgcccg tcagtgggca     9480

gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt     9540

gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt     9600

tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt     9660

cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc     9720

ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacgcccct ggctgcagta     9780

cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc     9840

ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cttgggcgct ggggccgccg     9900

cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat     9960

ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc    10020

gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg    10080

tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg    10140

acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat    10200

cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg    10260

gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg    10320

ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg    10380

tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag    10440

tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg    10500

ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct    10560

ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct    10620

tccatttcag gtgtcgtggg tttgggagct tgtatatcca ttttcggatc tgatcaagag    10680

acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc    10740

gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat    10800

gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg    10860

tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg    10920

ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta    10980

ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta    11040

tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc    11100

gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc    11160

gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg    11220

ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg    11280

ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt    11340

gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc    11400

ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc    11460

atcgccttct atcgccttct tgacgagttc ttctgattaa ttaacaggac tgaccgtgct    11520

acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg    11580

ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc    11640

caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac    11700

aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc    11760

ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct    11820

gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacagg atccactagt    11880

aacggccgcc agtgtgctgg aattccgagg tcgacggtat cgataagggc gcgccaaagc    11940

taactgtagg actgagtcta ttctaaactg aaagcctgga catctggagt accaggggga    12000

gatgacgtgt tacgggcttc cataaaagca gctggctttg aatggaagga gccaagaggc    12060

cagcacagga gcggattcgt cgctttcacg gccatcgagc cgaacctctc gcaagtccgt    12120

gagccgttaa ggaggccccc agtcccgacc cttcgcccca agcccctcgg ggtccccggg    12180

cctggtactc cttgccacac gggaggggcg cggaagccgg ggcggaggag gagccaaccc    12240

cgggctgggc tgagacccgc agaggaagac gctctaggga tttgtcccgg actagcgaga    12300

tggcaaggct gaggacggga ggctgattga gaggcgaagg tacaccctaa tctcaataca    12360

acctttggag ctaagccagc aatggtagag ggaagattct gcacgtccct tccaggcggc    12420

ctccccgtca ccaccccccc caacccgccc cgaccggagc tgagagtaat tcatacaaaa    12480

ggactcgccc ctgccttggg gaatcccagg gaccgtcgtt aaactcccac taacgtagaa    12540

cccagagatc gctgcgttcc cgccccctca cccgcccgct ctcgtcatca ctgaggtgga    12600

gaagagcatg cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc    12660

cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg    12720

gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa    12780

ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga    12840

acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct    12900

tgcgtgcctt gaattacttc cacgcccctg gctgcagtac gtgattcttg atcccgagct    12960

tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg    13020

tgcttgagtt gaggcctggc ttgggcgctg gggccgccgc gtgcgaatct ggtggcacct    13080

tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc    13140

tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc gatctgcaca    13200

ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat    13260

gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag    13320

ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg    13380

caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg    13440

ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca    13500

cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc    13560

gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt    13620

ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta    13680

ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt    13740

ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt    13800

gg                                                                   13802


<210>  7
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pMatrix


<220>
<221>  misc_feature
<222>  (1)..(42)
<223>  pMatrix

<400>  7
gtttaaacaa tattcctgca gggcagcccg gggagcggcc gc                          42


<210>  8
<211>  796
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pTriEx4neo vector construct sequence


<220>
<221>  misc_feature
<222>  (1)..(796)
<223>  pTriEx4neo vector construct sequence

<400>  8
gtttaaacgt catgatcgaa acatacaacc aaacttctcc ccgatctgcg gccactggac       60

tgcccatcag catgaaaatt tttatgtatt tacttactgt ttttcttatc acccagatga      120

ttgggtcagc actttttgct gtgtatcttc atagaaggtt ggacaagata gaagatgaaa      180

ggaatcttca tgaagatttt gtattcatga aaacgataca gagatgcaac acaggagaaa      240

gatccttatc cttactgaac tgtgaggaga ttaaaagcca gtttgaaggc tttgtgaagg      300

atataatgtt aaacaaagaa gagaaaaaag aagatgagga tcctgtagcc catgttgtag      360

caaaccctca agctgagggg cagctccagt ggctgaaccg ccgggccaat gccctcctgg      420

ccaatggcgt ggagctgaga gataaccagc tggtggtgcc atcagagggc ctgtacctca      480

tctactccca ggtcctcttc aagggccaag gctgcccctc cacccatgtg ctcctcaccc      540

acaccatcag ccgcatcgcc gtctcctacc agaccaaggt caacctcctc tctgccatca      600

agagcccctg ccagagggag accccagagg gggctgaggc caagccctgg tatgagccca      660

tctatctggg aggggtcttc cagctggaga agggtgaccg actcagcgct gagatcaatc      720

ggcccgacta tctcgacttt gccgagtctg ggcaggtcta ctttgggatc attgctctgt      780

gactcgaggc ggccgc                                                      796


<210>  9
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Forward (Fw) Primer (5' - 3') - Screening for site-specific 
       integration

<400>  9
ggaggattgg gaagacaata gc                                                22


<210>  10
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Reverse (Rev) Primer (5' - 3') - Screening for site-specific 
       integration

<400>  10
ctttcacagt cctgtacatc ttgt                                              24


<210>  11
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Filler amino acid sequence replacing a metalloproteinase 
       sensitive sequence of the human CD154 (Position 105 to 111)

<400>  11

Lys Lys Glu Asp Glu Asp Pro 
1               5           


<210>  12
<211>  7
<212>  PRT
<213>  Homo sapiens


<220>
<221>  PEPTIDE
<222>  (1)..(7)
<223>  Metalloproteinase sensitive sequence of the human CD154 (Position
       105 to 111)

<400>  12

Thr Lys Lys Glu Asn Ser Phe 
1               5           


