                         SEQUENCE LISTING

<110>  Instytut Biologii Doswiadczalnej im. Marcelego Nenckiego 
       Polska Akademia Nauk
 
<120>  Genetically encoded FRET-based MMP-9 activity biosensor and use 
       thereof

<130>  P30805PC00/MT

<150>  PL405046
<151>  2013-08-12

<160>  21    

<170>  PatentIn version 3.5

<210>  1
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus1 forward primer

<400>  1
ctggggccca gccggccaga tctcccggca tggtgagcaa gggcgagga                   49


<210>  2
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus1 reverse primer

<400>  2
tcctcgccct tgctcaccat gctagccttg tacagctcgt ccatgc                      46


<210>  3
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus2 forward primer

<400>  3
gcatggacga gctgtacaag gctagcatgg tgagcaaggg cgagga                      46


<210>  4
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus2 reverse primer

<400>  4
tcctcgccct tgctcaccat cttaagcttg tacagctcgt ccatgc                      46


<210>  5
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mTFP1 forward primer

<400>  5
gcatggacga gctgtacaag cttaagatgg tgagcaaggg cgagga                      46


<210>  6
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mTFP1 reverse primer

<400>  6
agatgagttt ttgttcgtcg acctgcagcc gcacttgtac agctcgtcca tgc              53


<210>  7
<211>  186
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus FP - mTFP1 linker (linker LN1)

<400>  7
cttaagggat ccccccgctc tctctctaag cttaaaggag gaaccggtgg aactggagga       60

accggtggaa ctggaggaac cggtggaact ggaggaaccg gtggaactgg aggaaccggt      120

ggaactggag gaaccggtgg aactggagga accggtggaa ctggaggaac cggtggaact      180

cttaag                                                                 186


<210>  8
<211>  183
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus FP - Venus FP linker (linker LN2)

<400>  8
gctagcggtg gtagcggtgg tagcggtgct agtggtggtt ctggttctag aggtggttct       60

ggttctagag gtggttctgg ttctagaggt ggttctggtt ctagaggtgg ttctggttct      120

agaggtggtt ctggttctag aggtggttct ggttctagag gtggttctgg ttctagagct      180

agc                                                                    183


<210>  9
<211>  84
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  alpha-helical linker with cleavage site

<400>  9
cttaaggagg aggagatcag agaggccttc agagtgttcc ccagaagcct gagcctgaga       60

cacgtgatga ccaacctgct taag                                              84


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

<220>
<223>  alpha-helical linker with cleavage site

<400>  10

Glu Glu Glu Ile Arg Glu Ala Phe Arg Val Phe Pro Arg Ser Leu Ser 
1               5                   10                  15      


Leu Arg His Val Met Thr Asn Leu 
            20                  


<210>  11
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mTFP1 forward primer

<400>  11
aagaacgggc ccatggtgag caagggcgag g                                      31


<210>  12
<211>  32
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mTFP1 reverse primer

<400>  12
aagaacagat ctcttgtaca gctcgtccat gc                                     32


<210>  13
<211>  62
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Venus FP - Venus FP linker (linker LN2)

<400>  13

Leu Lys Gly Ser Pro Arg Ser Leu Ser Lys Leu Lys Gly Gly Thr Gly 
1               5                   10                  15      


Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly 
            20                  25                  30          


Thr Gly Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Thr 
        35                  40                  45              


Gly Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Thr Leu Lys 
    50                  55                  60          


<210>  14
<211>  61
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Venus FP - Venus FP linker (linker LN2)

<400>  14

Ala Ser Gly Gly Ser Gly Gly Ser Gly Ala Ser Gly Gly Ser Gly Ser 
1               5                   10                  15      


Arg Gly Gly Ser Gly Ser Arg Gly Gly Ser Gly Ser Arg Gly Gly Ser 
            20                  25                  30          


Gly Ser Arg Gly Gly Ser Gly Ser Arg Gly Gly Ser Gly Ser Arg Gly 
        35                  40                  45              


Gly Ser Gly Ser Arg Gly Gly Ser Gly Ser Arg Ala Ser 
    50                  55                  60      


<210>  15
<211>  708
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mTFP1

<400>  15
atggtgagca agggcgagga gaccacaatg ggcgtaatca agcccgacat gaagatcaag       60

ctgaagatgg agggcaacgt gaatggccac gccttcgtga tcgagggcga gggcgagggc      120

aagccctacg acggcaccaa caccatcaac ctggaggtga aggagggagc ccccctgccc      180

ttctcctacg acattctgac caccgcgttc gcctacggca acagggcctt caccaagtac      240

cccgacgaca tccccaacta cttcaagcag tccttccccg agggctactc ttgggagcgc      300

accatgacct tcgaggacaa gggcatcgtg aaggtgaagt ccgacatctc catggaggag      360

gactccttca tctacgagat acacctcaag ggcgagaact tcccccccaa cggccccgtg      420

atgcagaaga agaccaccgg ctgggacgcc tccaccgaga ggatgtacgt gcgcgacggc      480

gtgctgaagg gcgacgtcaa gcacaagctg ctgctggagg gcggcggcca ccaccgcgtt      540

gacttcaaga ccatctacag ggccaagaag gcggtgaagc tgcccgacta tcactttgtg      600

gaccaccgca tcgagatcct gaaccacgac aaggactaca acaaggtgac cgtttacgag      660

agcgccgtgg cccgcaactc caccgacggc atggacgagc tgtacaag                   708


<210>  16
<211>  236
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  mTFP1

<400>  16

Met Val Ser Lys Gly Glu Glu Thr Thr Met Gly Val Ile Lys Pro Asp 
1               5                   10                  15      


Met Lys Ile Lys Leu Lys Met Glu Gly Asn Val Asn Gly His Ala Phe 
            20                  25                  30          


Val Ile Glu Gly Glu Gly Glu Gly Lys Pro Tyr Asp Gly Thr Asn Thr 
        35                  40                  45              


Ile Asn Leu Glu Val Lys Glu Gly Ala Pro Leu Pro Phe Ser Tyr Asp 
    50                  55                  60                  


Ile Leu Thr Thr Ala Phe Ala Tyr Gly Asn Arg Ala Phe Thr Lys Tyr 
65                  70                  75                  80  


Pro Asp Asp Ile Pro Asn Tyr Phe Lys Gln Ser Phe Pro Glu Gly Tyr 
                85                  90                  95      


Ser Trp Glu Arg Thr Met Thr Phe Glu Asp Lys Gly Ile Val Lys Val 
            100                 105                 110         


Lys Ser Asp Ile Ser Met Glu Glu Asp Ser Phe Ile Tyr Glu Ile His 
        115                 120                 125             


Leu Lys Gly Glu Asn Phe Pro Pro Asn Gly Pro Val Met Gln Lys Lys 
    130                 135                 140                 


Thr Thr Gly Trp Asp Ala Ser Thr Glu Arg Met Tyr Val Arg Asp Gly 
145                 150                 155                 160 


Val Leu Lys Gly Asp Val Lys His Lys Leu Leu Leu Glu Gly Gly Gly 
                165                 170                 175     


His His Arg Val Asp Phe Lys Thr Ile Tyr Arg Ala Lys Lys Ala Val 
            180                 185                 190         


Lys Leu Pro Asp Tyr His Phe Val Asp His Arg Ile Glu Ile Leu Asn 
        195                 200                 205             


His Asp Lys Asp Tyr Asn Lys Val Thr Val Tyr Glu Ser Ala Val Ala 
    210                 215                 220                 


Arg Asn Ser Thr Asp Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235     


<210>  17
<211>  717
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Venus

<400>  17
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac       60

ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac      120

ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc      180

ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag      240

cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc      300

ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg      360

gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac      420

aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac      480

ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc      540

gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac      600

tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc      660

ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaag         717


<210>  18
<211>  239
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Venus FP

<400>  18

Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 
1               5                   10                  15      


Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 
            20                  25                  30          


Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Leu Ile 
        35                  40                  45              


Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 
    50                  55                  60                  


Leu Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys 
65                  70                  75                  80  


Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 
                85                  90                  95      


Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 
            100                 105                 110         


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


Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 
    130                 135                 140                 


Asn Tyr Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn 
145                 150                 155                 160 


Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Gly 
                165                 170                 175     


Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 
            180                 185                 190         


Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Tyr Gln Ser Lys Leu 
        195                 200                 205             


Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 
    210                 215                 220                 


Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235                 


<210>  19
<211>  5325
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Plasmid pDisplay

<400>  19
gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag       60

ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct      120

gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc      180

caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg      240

cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat      300

ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca      360

tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc      420

gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga      480

gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat      540

tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc      600

taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga      660

cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg      720

cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt      780

tccaggttcc actggtgact atccatatga tgttccagat tatgctgggg cccagccggc      840

cagatctccc gggatccgcg gctgcaggtc gacgaacaaa aactcatctc agaagaggat      900

ctgaatgctg tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt      960

aaggtggtgg tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc     1020

atcctcatca tgctttggca gaagaagcca cgttaggcgg ccgctcgaga tcagcctcga     1080

ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc     1140

tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc     1200

tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt     1260

gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa     1320

gaaccagtgg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg     1380

tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc     1440

cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga     1500

aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct     1560

cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg     1620

gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag     1680

ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat     1740

cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac     1800

aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac     1860

tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc     1920

ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt     1980

tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc     2040

ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg     2100

agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca     2160

atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct     2220

gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg     2280

gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga     2340

accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc     2400

gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta     2460

gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc     2520

cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt     2580

gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga     2640

actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa     2700

gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca     2760

acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa     2820

agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat     2880

cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct     2940

gatgctcttg atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga     3000

tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc     3060

gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat     3120

cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga     3180

gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt     3240

gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg     3300

ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc     3360

cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca     3420

tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa     3480

aagcctcctc actacttctg gaatagctca gaggccgagg aggcggcctc ggcctctgca     3540

taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg     3600

ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt     3660

tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg     3720

agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac     3780

tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt caataaatca     3840

atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca     3900

cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag     3960

ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac     4020

ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc     4080

agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct     4140

agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc     4200

gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg     4260

cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc     4320

gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat     4380

tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag     4440

tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat     4500

aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg     4560

cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca     4620

cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga     4680

aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc     4740

ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata     4800

tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg     4860

ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc     4920

gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt     4980

ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc     5040

cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt     5100

agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt     5160

aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt     5220

gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa     5280

aaatttaacg cgaattttaa caaaatatta acgcttacaa tttac                     5325


<210>  20
<211>  150
<212>  DNA
<213>  Homo sapiens

<400>  20
gctgtgggcc aggacacgca ggaggtcatc gtggtgccac actccttgcc ctttaaggtg       60

gtggtgatct cagccatcct ggccctggtg gtgctcacca tcatctccct tatcatcctc      120

atcatgcttt ggcagaagaa gccacgttag                                       150


<210>  21
<211>  49
<212>  PRT
<213>  Homo sapiens

<400>  21

Ala Val Gly Gln Asp Thr Gln Glu Val Ile Val Val Pro His Ser Leu 
1               5                   10                  15      


Pro Phe Lys Val Val Val Ile Ser Ala Ile Leu Ala Leu Val Val Leu 
            20                  25                  30          


Thr Ile Ile Ser Leu Ile Ile Leu Ile Met Leu Trp Gln Lys Lys Pro 
        35                  40                  45              


Arg 
    


