                         SEQUENCE LISTING

<110>  Frommer, Wolf
       De Michele, Roberto
       Ast, Cynthia
 
<120>  TRANSMEMBRANE PROTEINS AS BIOSENSORS

<130>  056100-5085-WO

<150>  US 61/536,005
<151>  2011-09-18

<160>  32    

<170>  PatentIn version 3.5

<210>  1
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  1
aacgtctata tcatggcc                                                     18


<210>  2
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  2
tttttaccgg taccaccctt gtacagctcg tcca                                   34


<210>  3
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  3
tttttaccgg tggatctatg gtgagcaagg gcg                                    33


<210>  4
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  4
agttgtactc cagcttgtgc                                                   20


<210>  5
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  5
ttttttctag aaacgtctat atcatggcc                                         29


<210>  6
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  6
ttttttctag aagttgtact ccagcttgtg c                                      31


<210>  7
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  7
ttttttctag aatggtgagc aagggcgagg                                        30


<210>  8
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  8
ttttttctag acttgtacag ctcgtccatg                                        30


<210>  9
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  9
ttttttctag aaagggcgag gagctgttca                                        30


<210>  10
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  10
ttttttctag acttgtacag ctcgtccatg                                        30


<210>  11
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  11
ggtcctcgtc gtggtcggtt cgagaaatct cataacgtct atatcaag                    48


<210>  12
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  12
ggtcctcgtc gtggtcggtt cgagaaaggt ggttctcata acgtctatat caag             54


<210>  13
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  13
ggtcctcgtc gtggtcggtt cgagaaaggt ggtggtggtt ctcataacgt ctatatcaag       60


<210>  14
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  14
ggtcctcgtc gtggtcggtt cgagaaactc gagaacgtct atatcaag                    48


<210>  15
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  15
ggtcctcgtc gtggtcggtt cgagaaaggt ggtctcgaga acgtctatat caag             54


<210>  16
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  16
ggtcctcgtc gtggtcggtt cgagaaaggt ggtggtggtc tcgagaacgt ctatatcaag       60


<210>  17
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  17
gtggccgcgc agagcaatag cgcgaccacc gttgtactcc agcttg                      46


<210>  18
<211>  52
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  18
gtggccgcgc agagcaatag cgcgaccacc tcctccgttg tactccagct tg               52


<210>  19
<211>  52
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  19
gtggccgcgc agagcaatag cgcgaccacc attaaagttg tactccagct tg               52


<210>  20
<211>  52
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  20
gtggccgcgc agagcaatag cgcgaccacc tcttgtgttg tactccagct tg               52


<210>  21
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer


<220>
<221>  misc_feature
<222>  (28)..(33)
<223>  n is a, c, g, or t

<400>  21
ggtcctcgtc gtggtcggtt cgagaaannn nnnaacgtct atatcaag                    48


<210>  22
<211>  52
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  22
gtggccgcgc agagcaatag cgcgaccacc attaaagttg tactccagct tg               52


<210>  23
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ER targeting peptide

<400>  23

His Lys Thr Met Leu Pro Leu Pro Leu Ile Pro Ser Leu Leu Leu Ser 
1               5                   10                  15      


Leu Ser Ser Ala Glu Phe 
            20          


<210>  24
<211>  15
<212>  PRT
<213>  Nicotiana tabacum

<400>  24

Gln Pro Ser Leu Lys Arg Met Lys Ile Gln Pro Ser Ser Gln Pro 
1               5                   10                  15  


<210>  25
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  N-terminal myristate attachment sequence

<400>  25

Met Gly Ser Ser Lys Ser Lys 
1               5           


<210>  26
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SV-40 large T-antigen nuclear localisation sequence

<400>  26

Pro Lys Lys Lys Arg Lys Val 
1               5           


<210>  27
<211>  28
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  N-terminal targeting sequence

<400>  27

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


Arg Arg Leu Pro Val Pro Arg Ala Lys Ile Ser Leu 
            20                  25              


<210>  28
<211>  743
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Fusion protein

<400>  28

Met Ser Gly Ala Ile Thr Cys Ser Ala Ala Asp Leu Ala Thr Leu Leu 
1               5                   10                  15      


Gly Pro Asn Ala Thr Ala Ala Ala Asp Tyr Ile Cys Gly Gln Leu Gly 
            20                  25                  30          


Thr Val Asn Asn Lys Phe Thr Asp Ala Ala Phe Ala Ile Asp Asn Thr 
        35                  40                  45              


Tyr Leu Leu Phe Ser Ala Tyr Leu Val Phe Ala Met Gln Leu Gly Phe 
    50                  55                  60                  


Ala Met Leu Cys Ala Gly Ser Val Arg Ala Lys Asn Thr Met Asn Ile 
65                  70                  75                  80  


Met Leu Thr Asn Val Leu Asp Ala Ala Ala Gly Gly Leu Phe Tyr Tyr 
                85                  90                  95      


Leu Phe Gly Tyr Ala Phe Ala Phe Gly Gly Ser Ser Glu Gly Phe Ile 
            100                 105                 110         


Gly Arg His Asn Phe Ala Leu Arg Asp Phe Pro Thr Pro Thr Ala Asp 
        115                 120                 125             


Tyr Ser Phe Phe Leu Tyr Gln Trp Ala Phe Ala Ile Ala Ala Ala Gly 
    130                 135                 140                 


Ile Thr Ser Gly Ser Ile Ala Glu Arg Thr Gln Phe Val Ala Tyr Leu 
145                 150                 155                 160 


Ile Tyr Ser Ser Phe Leu Thr Gly Phe Val Tyr Pro Val Val Ser His 
                165                 170                 175     


Trp Phe Trp Ser Pro Asp Gly Trp Ala Ser Pro Phe Arg Ser Ala Asp 
            180                 185                 190         


Asp Arg Leu Phe Ser Thr Gly Ala Ile Asp Phe Ala Gly Ser Gly Val 
        195                 200                 205             


Val His Met Val Gly Gly Ile Ala Gly Leu Trp Gly Ala Leu Ile Glu 
    210                 215                 220                 


Gly Pro Arg Arg Gly Arg Phe Glu Lys Leu Glu Asn Val Tyr Ile Lys 
225                 230                 235                 240 


Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His 
                245                 250                 255     


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


Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu 
        275                 280                 285             


Ser Val Gln Ser Lys Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His 
    290                 295                 300                 


Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met 
305                 310                 315                 320 


Asp Glu Leu Tyr Lys Gly Gly Thr Gly Gly Ser Met Val Ser Lys Gly 
                325                 330                 335     


Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly 
            340                 345                 350         


Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp 
        355                 360                 365             


Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys 
    370                 375                 380                 


Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val 
385                 390                 395                 400 


Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe 
                405                 410                 415     


Lys Ser Ala Met Pro Glu Gly Tyr Ile Gln Glu Arg Thr Ile Phe Phe 
            420                 425                 430         


Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly 
        435                 440                 445             


Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu 
    450                 455                 460                 


Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Phe Asn Gly Gly 
465                 470                 475                 480 


Arg Ala Ile Ala Leu Arg Gly His Ser Ala Ser Leu Val Val Leu Gly 
                485                 490                 495     


Thr Phe Leu Leu Trp Phe Gly Trp Tyr Gly Phe Asn Pro Gly Ser Phe 
            500                 505                 510         


Thr Lys Ile Leu Val Pro Tyr Asn Ser Gly Ser Asn Tyr Gly Gln Trp 
        515                 520                 525             


Ser Gly Ile Gly Arg Thr Ala Val Asn Thr Thr Leu Ser Gly Cys Thr 
    530                 535                 540                 


Ala Ala Leu Thr Thr Leu Phe Gly Lys Arg Leu Leu Ser Gly His Trp 
545                 550                 555                 560 


Asn Val Thr Asp Val Cys Asn Gly Leu Leu Gly Gly Phe Ala Ala Ile 
                565                 570                 575     


Thr Ala Gly Cys Ser Val Val Glu Pro Trp Ala Ala Ile Val Cys Gly 
            580                 585                 590         


Phe Met Ala Ser Val Val Leu Ile Gly Cys Asn Lys Leu Ala Glu Leu 
        595                 600                 605             


Val Gln Tyr Asp Asp Pro Leu Glu Ala Ala Gln Leu His Gly Gly Cys 
    610                 615                 620                 


Gly Ala Trp Gly Leu Ile Phe Val Gly Leu Phe Ala Lys Glu Lys Tyr 
625                 630                 635                 640 


Leu Asn Glu Val Tyr Gly Ala Thr Pro Gly Arg Pro Tyr Gly Leu Phe 
                645                 650                 655     


Met Gly Gly Gly Gly Lys Leu Leu Gly Ala Gln Leu Val Gln Ile Leu 
            660                 665                 670         


Val Ile Val Gly Trp Val Ser Ala Thr Met Gly Thr Leu Phe Phe Ile 
        675                 680                 685             


Leu Lys Arg Leu Asn Leu Leu Arg Ile Ser Glu Gln His Glu Met Gln 
    690                 695                 700                 


Gly Met Asp Met Thr Arg His Gly Gly Phe Ala Tyr Ile Tyr His Asp 
705                 710                 715                 720 


Asn Asp Asp Glu Ser His Arg Val Asp Pro Gly Ser Pro Phe Pro Arg 
                725                 730                 735     


Ser Ala Thr Pro Pro Arg Val 
            740             


<210>  29
<211>  2229
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of fusion protein

<400>  29
atgtcaggag caataacatg ctctgcggcc gatctcgcca ccctacttgg ccccaacgcc       60

acggcggcgg ccgactacat ttgcggccaa ttaggcaccg ttaacaacaa gttcaccgat      120

gcagccttcg ccatagacaa cacctacctc ctcttctctg cctaccttgt cttcgccatg      180

cagctcggct tcgctatgct ttgtgctggt tctgttagag ccaagaatac gatgaacatc      240

atgcttacca atgtccttga cgctgcagcc ggaggactct tctactatct ctttggttac      300

gcctttgcct ttggaggatc ctccgaaggg ttcattggaa gacacaactt tgctcttaga      360

gactttccga ctcccacagc tgattactct ttcttcctct accaatgggc gttcgcaatc      420

gcggccgctg gaatcacaag tggttcgatc gcagagagga ctcagttcgt ggcttacttg      480

atatactctt ctttcttaac cggatttgtt tacccggttg tctctcactg gttttggtcc      540

ccggatggat gggccagtcc ctttcgttca gcggatgatc gtttgtttag caccggagcc      600

attgactttg ctggctccgg tgttgttcac atggttggtg gcatagcagg tttatggggt      660

gctcttattg aaggtcctcg tcgtggtcgg ttcgagaaac tcgagaacgt ctatatcaag      720

gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac      780

ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg      840

ctgctgcccg acaaccacta cctgagcgtc cagtccaagc tgagcaaaga ccccaacgag      900

aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg      960

gacgagctgt acaagggtgg taccggtgga tctatggtga gcaagggcga ggagctgttc     1020

accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc     1080

gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc     1140

accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg     1200

cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg     1260

cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc     1320

cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc     1380

gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt taatggtggt     1440

cgcgctattg ctctgcgcgg ccactctgcc tcgctagtag tcttaggaac cttcctccta     1500

tggtttggat ggtatggttt caaccccggt tccttcacta agatactcgt tccgtataat     1560

tctggttcca actacggcca atggagcgga atcggccgta cagcggttaa caccacactc     1620

tcaggatgca cagcagctct aaccacactc tttggtaaac gtctcctatc aggccactgg     1680

aacgtaacgg acgtttgcaa cgggttactc ggtgggtttg cggccataac cgcaggttgc     1740

tccgtcgtag agccatgggc agcgattgtg tgcggcttca tggcttctgt cgtccttatc     1800

ggatgcaaca agctcgcgga gcttgtacaa tatgatgatc cactcgaggc agcccaacta     1860

catggagggt gtggcgcgtg ggggttgata ttcgtaggat tgtttgccaa agagaagtat     1920

ctaaacgagg tttatggcgc caccccggga aggccatatg gactatttat gggcggagga     1980

gggaagctgt tgggagcaca attggttcaa atacttgtga ttgtaggatg ggttagtgcc     2040

acaatgggaa cactcttctt catcctcaaa aggctcaatc tgcttaggat ctcggagcag     2100

catgaaatgc aagggatgga tatgacacgt cacggtggct ttgcttatat ctaccatgat     2160

aatgatgatg agtctcatag agtggatcct ggatctcctt tccctcgatc agctactcct     2220

cctcgcgtt                                                             2229


<210>  30
<211>  2240
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence of fusion protein

<400>  30
atgtcaggag caataacatg ctctgcggcc gatctcgcca ccctacttgg ccccaacgcc       60

acggcggcgg ccgactacat ttgcggccaa ttaggcaccg ttaacaacaa gttcaccgat      120

gcagccttcg ccatagacaa cacctacctc ctcttctctg cctaccttgt cttcgccatg      180

cagctcggct tcgctatgct ttgtgctggt tctgttagag ccaagaatac gatgaacatc      240

atgcttacca atgtccttga cgctgcagcc ggaggactct tctactatct ctttggttac      300

gcctttgcct ttggaggatc ctccgaaggg ttcattggaa gacacaactt tgctcttaga      360

gactttccga ctcccacagc tgattactct ttcttcctct accaatgggc gttcgcaatc      420

gcggccgctg gaatcacaag tggttcgatc gcagagagga ctcagttcgt ggcttacttg      480

atatactctt ctttcttaac cggatttgtt tacccggttg tctctcactg gttttggtcc      540

ccggatggat gggccagtcc ctttcgttca gcggatgatc gtttgtttag caccggagcc      600

attgactttg ctggctccgg tgttgttcac atggttggtg gcatagcagg tttatggggt      660

gctcttattg aaggtcctcg tcgtggtcgg ttcgagaaag gtggtctcga gaacgtaacg      720

tctatatcaa ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca      780

acatcgagga cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg      840

acggccccgt gctgctgccc gacaaccact acctgagcgt ccagtccaag ctgagcaaag      900

accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca      960

ctctcggcat ggacgagctg tacaagggtg gtaccggtgg atctatggtg agcaagggcg     1020

aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc     1080

acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga     1140

agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga     1200

cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca     1260

agtccgccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca     1320

actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc     1380

tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact     1440

ttaatggtgg tcgcgctatt gctctgcgcg gccactctgc ctcgctagta gtcttaggaa     1500

ccttcctcct atggtttgga tggtatggtt tcaaccccgg ttccttcact aagatactcg     1560

ttccgtataa ttctggttcc aactacggcc aatggagcgg aatcggccgt acagcggtta     1620

acaccacact ctcaggatgc acagcagctc taaccacact ctttggtaaa cgtctcctat     1680

caggccactg gaacgtaacg gacgtttgca acgggttact cggtgggttt gcggccataa     1740

ccgcaggttg ctccgtcgta gagccatggg cagcgattgt gtgcggcttc atggcttctg     1800

tcgtccttat cggatgcaac aagctcgcgg agcttgtaca atatgatgat ccactcgagg     1860

cagcccaact acatggaggg tgtggcgcgt gggggttgat attcgtagga ttgtttgcca     1920

aagagaagta tctaaacgag gtttatggcg ccaccccggg aaggccatat ggactattta     1980

tgggcggagg agggaagctg ttgggagcac aattggttca aatacttgtg attgtaggat     2040

gggttagtgc cacaatggga acactcttct tcatcctcaa aaggctcaat ctgcttagga     2100

tctcggagca gcatgaaatg caagggatgg atatgacacg tcacggtggc tttgcttata     2160

tctaccatga taatgatgat gagtctcata gagtggatcc tggatctcct ttccctcgat     2220

cagctactcc tcctcgcgtt                                                 2240


<210>  31
<211>  241
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  cpGFP portion

<400>  31

Asn Val Tyr Ile Lys Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn 
1               5                   10                  15      


Phe Lys Ile Arg His Asn Ile Glu Asp Gly Gly Val Gln Leu Ala Tyr 
            20                  25                  30          


His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro 
        35                  40                  45              


Asp Asn His Tyr Leu Ser Val Gln Ser Lys Leu Ser Lys Asp Pro Asn 
    50                  55                  60                  


Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly 
65                  70                  75                  80  


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


Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 
            100                 105                 110         


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


Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 
    130                 135                 140                 


Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 
145                 150                 155                 160 


Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 
                165                 170                 175     


Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Ile Gln Glu 
            180                 185                 190         


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


Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 
    210                 215                 220                 


Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 
225                 230                 235                 240 


Asn 
    


<210>  32
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker

<400>  32

Gly Gly Thr Gly Gly Ser 
1               5       


