                         SEQUENCE LISTING

<110>  Horton, Robert
       Michaud, Gregory
       Riddle, Steven
       Robers, Matthew
       Vogel, Kurt
 
<120>  KINASE AND UBIQUINATION ASSAYS

<130>  IVGN 924.1 PCT

<140>  UNKNOWN
<141>  2007-07-20

<150>  60/832,114
<151>  2006-07-21

<160>  18    

<170>  PatentIn version 3.3

<210>  1
<211>  1713
<212>  DNA
<213>  Artificial

<220>
<223>  GFP-IkBalpha Coding Sequence

<400>  1
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac     60

ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac    120

ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc    180

ctcgtgacca ccttcaccta cggcgtgcag 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 cagccacaag gtctatatca ccgccgacaa gcagaagaac    480

ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc    540

gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac    600

tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc    660

ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc    720

tcgagcccat caacaagttt gtacaaaaaa gcaggcacca tgttccaggc ggccgagcgc    780

ccccaggagt gggccatgga gggcccccgc gacgggctga agaaggagcg gctactggac    840

gaccgccacg acagcggcct ggactccatg aaagacgagg agtacgagca gatggtcaag    900

gagctgcagg agatccgcct cgagccgcag gaggtgccgc gcggctcgga gccctggaag    960

cagcagctca ccgaggacgg ggactcgttc ctgcacttgg ccatcatcca tgaagaaaag   1020

gcactgacca tggaagtgat ccgccaggtg aagggagacc tggccttcct caacttccag   1080

aacaacctgc agcagactcc actccacttg gctgtgatca ccaaccagcc agaaattgct   1140

gaggcacttc tgggagctgg ctgtgatcct gagctccgag actttcgagg aaataccccc   1200

ctacaccttg cctgtgagca gggctgcctg gccagcgtgg gagtcctgac tcagtcctgc   1260

accaccccgc acctccactc catcctgaag gctaccaact acaatggcca cacgtgtcta   1320

cacttagcct ctatccatgg ctacctgggc atcgtggagc ttttggtgtc cttgggtgct   1380

gatgtcaatg ctcaggagcc ctgtaatggc cggactgccc ttcacctcgc agtggacctg   1440

caaaatcctg acctggtgtc actcctgttg aagtgtgggg ctgatgtcaa cagagttacc   1500

taccagggct attctcccta ccagctcacc tggggccgcc caagcacccg gatacagcag   1560

cagctgggcc agctgacact agaaaacctt cagatgctgc cagagagtga ggatgaggag   1620

agctatgaca cagagtcaga gttcacggag ttcacagagg acgagctgcc ctatgatgac   1680

tgtgtgtttg gaggccagcg tctgacgtta tag                                1713


<210>  2
<211>  369
<212>  PRT
<213>  Artificial

<220>
<223>  Synthetic amino acid

<400>  2

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr 
1               5                   10                  15      


Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp 
            20                  25                  30          


Arg Trp Gly Ser Glu Phe Ala Thr Met Val Ser Lys Gly Glu Glu Leu 
        35                  40                  45              


Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn 
    50                  55                  60                  


Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr 
65                  70                  75                  80  


Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val 
                85                  90                  95      


Pro Trp Pro Thr Leu Val Thr Thr Phe Gly Tyr Gly Val Gln Cys Phe 
            100                 105                 110         


Ala Arg Tyr Pro Asp His Met Arg Gln His Asp Phe Phe Lys Ser Ala 
        115                 120                 125             


Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp 
    130                 135                 140                 


Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu 
145                 150                 155                 160 


Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn 
                165                 170                 175     


Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr 
            180                 185                 190         


Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Ile 
        195                 200                 205             


Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln 
    210                 215                 220                 


Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His 
225                 230                 235                 240 


Tyr Leu Ser Tyr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg 
                245                 250                 255     


Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu 
            260                 265                 270         


Gly Met Asp Glu Leu Tyr Lys Leu Glu Thr Asp Gln Thr Ser Leu Tyr 
        275                 280                 285             


Lys Lys Ala Gly Thr Met Gln Ile Phe Val Lys Thr Leu Thr Gly Lys 
    290                 295                 300                 


Thr Ile Thr Leu Glu Val Glu Pro Ser Asp Thr Ile Glu Asn Val Lys 
305                 310                 315                 320 


Ala Lys Ile Gln Asp Lys Glu Gly Ile Pro Pro Asp Gln Gln Arg Leu 
                325                 330                 335     


Ile Phe Ala Gly Lys Gln Leu Glu Asp Gly Arg Thr Leu Ser Asp Tyr 
            340                 345                 350         


Asn Ile Gln Lys Glu Ser Thr Leu His Leu Val Leu Arg Leu Arg Gly 
        355                 360                 365             


Gly 
    


<210>  3
<211>  4
<212>  PRT
<213>  Artificial

<220>
<223>  Consensus sequence


<220>
<221>  misc_feature
<222>  (1)..(1)
<223>  Xaa = large hydrophobic amino acid

<220>
<221>  misc_feature
<222>  (3)..(3)
<223>  Xaa = any amino acid

<400>  3

Xaa Lys Xaa Glu 
1               


<210>  4
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  PTK labeled tracer


<220>
<221>  misc_feature
<222>  (4)..(4)
<223>  Xaa = phosphorylated tyrosine

<400>  4

Ala Asp Glu Xaa Leu Ile Pro Gln Gln Ser 
1               5                   10  


<210>  5
<211>  11
<212>  PRT
<213>  Artificial

<220>
<223>  STK labeled tracer


<220>
<221>  misc_feature
<222>  (7)..(7)
<223>  Xaa = phosphorylated serine

<400>  5

Gly Arg Pro Arg Thr Ser Xaa Phe Ala Glu Gly 
1               5                   10      


<210>  6
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  Peptide competitor


<220>
<221>  misc_feature
<222>  (4)..(4)
<223>  Xaa = phosphorylated tyrosine

<400>  6

Ala Asp Glu Xaa Leu Ile Pro Gln Gln Ser 
1               5                   10  


<210>  7
<211>  14
<212>  PRT
<213>  Artificial

<220>
<223>  Tracer peptide


<220>
<221>  misc_feature
<222>  (11)..(11)
<223>  Xaa = phosphorylated serine

<400>  7

Leu Arg Arg Glu Ile Leu Ser Arg Arg Pro Xaa Tyr Arg Lys 
1               5                   10                  


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

<220>
<223>  Tracer peptide


<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  Xaa = phosphorylated serine

<400>  8

Arg Glu Ile Leu Ser Arg Arg Pro Xaa Tyr Arg Lys 
1               5                   10          


<210>  9
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  Tracer peptide


<220>
<221>  misc_feature
<222>  (7)..(7)
<223>  Xaa = phosphorylated serine

<400>  9

Ile Leu Ser Arg Arg Pro Xaa Tyr Arg Lys 
1               5                   10  


<210>  10
<211>  14
<212>  PRT
<213>  Artificial 

<220>
<223>  Tracer protein


<220>
<221>  misc_feature
<222>  (11)..(11)
<223>  Xaa = phosphorylated serine

<400>  10

Leu Arg Arg Glu Ile Leu Ser Arg Arg Pro Xaa Tyr Arg Lys 
1               5                   10                  


<210>  11
<211>  14
<212>  PRT
<213>  Artificial

<220>
<223>  PKA substrate

<400>  11

Leu Arg Arg Glu Ile Leu Ser Arg Arg Pro Ser Tyr Arg Lys 
1               5                   10                  


<210>  12
<211>  11
<212>  PRT
<213>  Artificial

<220>
<223>  Labeled tracer


<220>
<221>  misc_feature
<222>  (5)..(5)
<223>  Xaa = phosphorylated tyrosine

<400>  12

Cys Ala Asp Glu Xaa Leu Ile Pro Gln Gln Ser 
1               5                   10      


<210>  13
<211>  6
<212>  PRT
<213>  Artificial

<220>
<223>  luminescent tracer

<400>  13

His His His His His His 
1               5       


<210>  14
<211>  9
<212>  PRT
<213>  Artificial

<220>
<223>  Tagged peptide

<400>  14

Lys Gly Gly His His His His His His 
1               5                   


<210>  15
<211>  371
<212>  PRT
<213>  Artificial

<220>
<223>  DUB substrate

<400>  15

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr 
1               5                   10                  15      


Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp 
            20                  25                  30          


Arg Trp Gly Ser Glu Phe Ala Thr Met Val Ser Lys Gly Glu Glu Leu 
        35                  40                  45              


Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn 
    50                  55                  60                  


Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr 
65                  70                  75                  80  


Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val 
                85                  90                  95      


Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe 
            100                 105                 110         


Ala Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala 
        115                 120                 125             


Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp 
    130                 135                 140                 


Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu 
145                 150                 155                 160 


Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn 
                165                 170                 175     


Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Lys Val Tyr 
            180                 185                 190         


Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Thr 
        195                 200                 205             


Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln 
    210                 215                 220                 


Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His 
225                 230                 235                 240 


Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg 
                245                 250                 255     


Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu 
            260                 265                 270         


Gly Met Asp Glu Leu Tyr Lys Leu Glu Thr Asp Gln Thr Ser Leu Tyr 
        275                 280                 285             


Lys Lys Ala Gly Thr Met Gln Ile Phe Val Lys Thr Leu Thr Gly Lys 
    290                 295                 300                 


Thr Ile Thr Leu Glu Val Glu Pro Ser Asp Thr Ile Glu Asn Val Lys 
305                 310                 315                 320 


Ala Lys Ile Gln Asp Lys Glu Gly Ile Pro Pro Asp Gln Gln Arg Leu 
                325                 330                 335     


Ile Phe Ala Gly Lys Gln Leu Glu Asp Gly Arg Thr Leu Ser Asp Tyr 
            340                 345                 350         


Asn Ile Gln Lys Glu Ser Thr Leu His Leu Val Leu Arg Leu Arg Gly 
        355                 360                 365             


Gly Ala Cys 
    370     


<210>  16
<211>  379
<212>  PRT
<213>  Artificial

<220>
<223>  DUB substrate

<400>  16

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr 
1               5                   10                  15      


Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp 
            20                  25                  30          


Arg Trp Gly Ser Glu Phe Ala Thr Met Val Ser Lys Gly Glu Glu Leu 
        35                  40                  45              


Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn 
    50                  55                  60                  


Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr 
65                  70                  75                  80  


Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val 
                85                  90                  95      


Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe 
            100                 105                 110         


Ala Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala 
        115                 120                 125             


Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp 
    130                 135                 140                 


Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu 
145                 150                 155                 160 


Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn 
                165                 170                 175     


Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Lys Val Tyr 
            180                 185                 190         


Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Thr 
        195                 200                 205             


Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln 
    210                 215                 220                 


Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His 
225                 230                 235                 240 


Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg 
                245                 250                 255     


Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu 
            260                 265                 270         


Gly Met Asp Glu Leu Tyr Lys Leu Glu Thr Asp Gln Thr Ser Leu Tyr 
        275                 280                 285             


Lys Lys Ala Gly Thr Met Gln Ile Phe Val Lys Thr Leu Thr Gly Lys 
    290                 295                 300                 


Thr Ile Thr Leu Glu Val Glu Pro Ser Asp Thr Ile Glu Asn Val Lys 
305                 310                 315                 320 


Ala Lys Ile Gln Asp Lys Glu Gly Ile Pro Pro Asp Gln Gln Arg Leu 
                325                 330                 335     


Ile Phe Ala Gly Lys Gln Leu Glu Asp Gly Arg Thr Leu Ser Asp Tyr 
            340                 345                 350         


Asn Ile Gln Lys Glu Ser Thr Leu His Leu Val Leu Arg Leu Arg Gly 
        355                 360                 365             


Gly Phe Phe Gly Val Gly Gly Glu Gly Ala Cys 
    370                 375                 


<210>  17
<211>  372
<212>  PRT
<213>  Artificial

<220>
<223>  Fusion protein

<400>  17

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr 
1               5                   10                  15      


Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp 
            20                  25                  30          


Arg Trp Gly Ser Glu Phe Ala Thr Met Val Ser Lys Gly Glu Glu Leu 
        35                  40                  45              


Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn 
    50                  55                  60                  


Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr 
65                  70                  75                  80  


Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val 
                85                  90                  95      


Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe 
            100                 105                 110         


Ala Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala 
        115                 120                 125             


Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp 
    130                 135                 140                 


Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu 
145                 150                 155                 160 


Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn 
                165                 170                 175     


Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Lys Val Tyr 
            180                 185                 190         


Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Thr 
        195                 200                 205             


Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln 
    210                 215                 220                 


Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His 
225                 230                 235                 240 


Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg 
                245                 250                 255     


Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu 
            260                 265                 270         


Gly Met Asp Glu Leu Tyr Lys Leu Glu Thr Asp Gln Thr Ser Leu Tyr 
        275                 280                 285             


Lys Lys Ala Gly Ser Met Thr Ala Lys Met Glu Thr Thr Phe Tyr Asp 
    290                 295                 300                 


Asp Ala Leu Asn Ala Ser Phe Leu Pro Ser Glu Ser Gly Pro Tyr Gly 
305                 310                 315                 320 


Tyr Ser Asn Pro Lys Ile Leu Lys Gln Ser Met Thr Leu Asn Leu Ala 
                325                 330                 335     


Asp Pro Val Gly Ser Leu Lys Pro His Leu Arg Ala Lys Asn Ser Asp 
            340                 345                 350         


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


Leu Glu Arg Leu 
    370         


<210>  18
<211>  371
<212>  PRT
<213>  Artificial

<220>
<223>  Fusion protein

<400>  18

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr 
1               5                   10                  15      


Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp 
            20                  25                  30          


Arg Trp Gly Ser Glu Phe Ala Thr Met Val Ser Lys Gly Glu Glu Leu 
        35                  40                  45              


Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn 
    50                  55                  60                  


Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr 
65                  70                  75                  80  


Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val 
                85                  90                  95      


Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe 
            100                 105                 110         


Ala Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala 
        115                 120                 125             


Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp 
    130                 135                 140                 


Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu 
145                 150                 155                 160 


Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn 
                165                 170                 175     


Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Lys Val Tyr 
            180                 185                 190         


Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Thr 
        195                 200                 205             


Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln 
    210                 215                 220                 


Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His 
225                 230                 235                 240 


Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg 
                245                 250                 255     


Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu 
            260                 265                 270         


Gly Met Asp Glu Leu Tyr Lys Leu Glu Thr Asp Gln Thr Ser Leu Tyr 
        275                 280                 285             


Lys Lys Ala Gly Ser Met Ser Asp Asp Lys Pro Phe Leu Cys Thr Ala 
    290                 295                 300                 


Pro Gly Cys Gly Gln Arg Phe Thr Asn Glu Asp His Leu Ala Val His 
305                 310                 315                 320 


Lys His Lys His Glu Met Thr Leu Lys Phe Gly Pro Ala Arg Asn Asp 
                325                 330                 335     


Ser Val Ile Val Ala Asp Gln Thr Pro Thr Pro Thr Arg Phe Leu Lys 
            340                 345                 350         


Asn Cys Glu Glu Val Gly Leu Phe Asn Glu Leu Ala Ser Pro Phe Glu 
        355                 360                 365             


Asn Glu Phe 
    370     


