                               SEQUENCE LISTING

<110> THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
 
<120> SITE-SPECIFIC GENERATION OF PHOSPHORYLATED TYROSINES IN PROTEIN 

<130> UCSF039PCT

<140>
<141>

<150> 62/458,557
<151> 2017-02-13

<160> 3     

<170> PatentIn version 3.5

<210> 1
<211> 1365
<212> DNA
<213> Methanosarcina mazei

<220>
<223> mutant pyrrolysyl-tRNA synthetase 

<400> 1
atggataaaa agcctctgaa cactctgatt tctgcgaccg gtctgtggat gtcccgcacc       60

ggcaccatcc acaaaatcaa acaccatgaa gttagccgtt ccaaaatcta cattgaaatg      120

gcttgcggcg atcacctggt tgtcaacaac tcccgttctt ctcgtaccgc tcgcgcactg      180

cgccaccaca aatatcgcaa aacctgcaaa cgttgccgtg ttagcgatga agatctgaac      240

aaattcctga ccaaagctaa cgaggatcag acctccgtaa aagtgaaggt agtaagcgct      300

ccgacccgta ctaaaaaggc tatgccaaaa agcgtggccc gtgccccgaa acctctggaa      360

aacaccgagg cggctcaggc tcaaccatcc ggttctaaat tttctccggc gatcccagtg      420

tccacccaag aatctgtttc cgtaccagca agcgtgtcta ccagcattag cagcatttct      480

accggtgcta ccgcttctgc gctggtaaaa ggtaacacta acccgattac tagcatgtct      540

gcaccggtac aggcaagcgc cccagctctg actaaatccc agacggaccg tctggaggtg      600

ctgctgaacc caaaggatga aatctctctg aacagcggca agcctttccg tgagctggaa      660

agcgagctgc tgtctcgtcg taaaaaggat ctgcaacaga tctacgctga ggaacgcgag      720

aactatctgg gtaagctgga gcgcgaaatt actcgcttct tcgtggatcg cggtttcctg      780

gagatcaaat ctccgattct gattccgctg gaatacattg aacgtatggg catcgataat      840

gataccgaac tgtctaaaca gatcttccgt gtggataaaa acttctgtct gcgtccgatg      900

ctgtctccga acctgtacaa ctatatgcgt aaactggacc gtgccctgcc ggacccgatc      960

aaattgttcg agatcggtcc ttgctaccgt aaagagtccg acggtaaaga gcacctggaa     1020

gaattcacca tgctggcttt cggtcagatg ggtagcggtt gcacgcgtga aaacctggaa     1080

tccattatca ccgacttcct gaatcacctg ggtatcgatt tcaaaattgt tggtgacagc     1140

tgtatggtgt atggcgatac gctggatgtt atgcacggcg atctggagct gtcttccgca     1200

gttgtgggcc caatcccgct ggatcgtgag tggggtatcg acaaacctac gatcggtgcg     1260

ggttttggtc tggagcgtct gctgaaagta aaacacgact tcaagaacat caaacgtgct     1320

gcacgttccg agtcctatta caatggtatt tctactaacc tgtaa                     1365


<210> 2
<211> 454
<212> PRT
<213> Methanosarcina mazei

<220>
<223> mutant pyrrolysyl-tRNA synthetase 

<400> 2
Met Asp Lys Lys Pro Leu Asn Thr Leu Ile Ser Ala Thr Gly Leu Trp 
1               5                   10                  15      


Met Ser Arg Thr Gly Thr Ile His Lys Ile Lys His His Glu Val Ser 
            20                  25                  30          


Arg Ser Lys Ile Tyr Ile Glu Met Ala Cys Gly Asp His Leu Val Val 
        35                  40                  45              


Asn Asn Ser Arg Ser Ser Arg Thr Ala Arg Ala Leu Arg His His Lys 
    50                  55                  60                  


Tyr Arg Lys Thr Cys Lys Arg Cys Arg Val Ser Asp Glu Asp Leu Asn 
65                  70                  75                  80  


Lys Phe Leu Thr Lys Ala Asn Glu Asp Gln Thr Ser Val Lys Val Lys 
                85                  90                  95      


Val Val Ser Ala Pro Thr Arg Thr Lys Lys Ala Met Pro Lys Ser Val 
            100                 105                 110         


Ala Arg Ala Pro Lys Pro Leu Glu Asn Thr Glu Ala Ala Gln Ala Gln 
        115                 120                 125             


Pro Ser Gly Ser Lys Phe Ser Pro Ala Ile Pro Val Ser Thr Gln Glu 
    130                 135                 140                 


Ser Val Ser Val Pro Ala Ser Val Ser Thr Ser Ile Ser Ser Ile Ser 
145                 150                 155                 160 


Thr Gly Ala Thr Ala Ser Ala Leu Val Lys Gly Asn Thr Asn Pro Ile 
                165                 170                 175     


Thr Ser Met Ser Ala Pro Val Gln Ala Ser Ala Pro Ala Leu Thr Lys 
            180                 185                 190         


Ser Gln Thr Asp Arg Leu Glu Val Leu Leu Asn Pro Lys Asp Glu Ile 
        195                 200                 205             


Ser Leu Asn Ser Gly Lys Pro Phe Arg Glu Leu Glu Ser Glu Leu Leu 
    210                 215                 220                 


Ser Arg Arg Lys Lys Asp Leu Gln Gln Ile Tyr Ala Glu Glu Arg Glu 
225                 230                 235                 240 


Asn Tyr Leu Gly Lys Leu Glu Arg Glu Ile Thr Arg Phe Phe Val Asp 
                245                 250                 255     


Arg Gly Phe Leu Glu Ile Lys Ser Pro Ile Leu Ile Pro Leu Glu Tyr 
            260                 265                 270         


Ile Glu Arg Met Gly Ile Asp Asn Asp Thr Glu Leu Ser Lys Gln Ile 
        275                 280                 285             


Phe Arg Val Asp Lys Asn Phe Cys Leu Arg Pro Met Leu Ser Pro Asn 
    290                 295                 300                 


Leu Tyr Asn Tyr Met Arg Lys Leu Asp Arg Ala Leu Pro Asp Pro Ile 
305                 310                 315                 320 


Lys Leu Phe Glu Ile Gly Pro Cys Tyr Arg Lys Glu Ser Asp Gly Lys 
                325                 330                 335     


Glu His Leu Glu Glu Phe Thr Met Leu Ala Phe Gly Gln Met Gly Ser 
            340                 345                 350         


Gly Cys Thr Arg Glu Asn Leu Glu Ser Ile Ile Thr Asp Phe Leu Asn 
        355                 360                 365             


His Leu Gly Ile Asp Phe Lys Ile Val Gly Asp Ser Cys Met Val Tyr 
    370                 375                 380                 


Gly Asp Thr Leu Asp Val Met His Gly Asp Leu Glu Leu Ser Ser Ala 
385                 390                 395                 400 


Val Val Gly Pro Ile Pro Leu Asp Arg Glu Trp Gly Ile Asp Lys Pro 
                405                 410                 415     


Thr Ile Gly Ala Gly Phe Gly Leu Glu Arg Leu Leu Lys Val Lys His 
            420                 425                 430         


Asp Phe Lys Asn Ile Lys Arg Ala Ala Arg Ser Glu Ser Tyr Tyr Asn 
        435                 440                 445             


Gly Ile Ser Thr Asn Leu 
    450                 


<210> 3
<211> 419
<212> PRT
<213> Methanosarcina barkeri

<220>
<223> mutant pyrrolysyl-tRNA synthetase 

<400> 3
Met Asp Lys Lys Pro Leu Asp Val Leu Ile Ser Ala Thr Gly Leu Trp 
1               5                   10                  15      


Met Ser Arg Thr Gly Thr Leu His Lys Ile Lys His His Glu Val Ser 
            20                  25                  30          


Arg Ser Lys Ile Tyr Ile Glu Met Ala Cys Gly Asp His Leu Val Val 
        35                  40                  45              


Asn Asn Ser Arg Ser Cys Arg Thr Ala Arg Ala Phe Arg His His Lys 
    50                  55                  60                  


Tyr Arg Lys Thr Cys Lys Arg Cys Arg Val Ser Asp Glu Asp Ile Asn 
65                  70                  75                  80  


Asn Phe Leu Thr Arg Ser Thr Glu Ser Lys Asn Ser Val Lys Val Arg 
                85                  90                  95      


Val Val Ser Ala Pro Lys Val Lys Lys Ala Met Pro Lys Ser Val Ser 
            100                 105                 110         


Arg Ala Pro Lys Pro Leu Glu Asn Ser Val Ser Ala Lys Ala Ser Thr 
        115                 120                 125             


Asn Thr Ser Arg Ser Val Pro Ser Pro Ala Lys Ser Thr Pro Asn Ser 
    130                 135                 140                 


Ser Val Pro Ala Ser Ala Pro Ala Pro Ser Leu Thr Arg Ser Gln Leu 
145                 150                 155                 160 


Asp Arg Val Glu Ala Leu Leu Ser Pro Glu Asp Lys Ile Ser Leu Asn 
                165                 170                 175     


Met Ala Lys Pro Phe Arg Glu Leu Glu Pro Glu Leu Val Thr Arg Arg 
            180                 185                 190         


Lys Asn Asp Phe Gln Arg Leu Tyr Thr Asn Asp Arg Glu Asp Tyr Leu 
        195                 200                 205             


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


Leu Glu Ile Lys Ser Pro Ile Leu Ile Pro Ala Glu Tyr Val Glu Arg 
225                 230                 235                 240 


Met Gly Ile Asn Asn Asp Thr Glu Leu Ser Lys Gln Ile Phe Arg Val 
                245                 250                 255     


Asp Lys Asn Leu Cys Leu Arg Pro Met Leu Ser Pro Thr Leu Tyr Asn 
            260                 265                 270         


Tyr Met Arg Lys Leu Asp Arg Ile Leu Pro Gly Pro Ile Lys Leu Phe 
        275                 280                 285             


Glu Val Gly Pro Cys Tyr Arg Lys Glu Ser Asp Gly Lys Glu His Leu 
    290                 295                 300                 


Glu Glu Phe Thr Met Val Ala Phe Gly Gln Met Gly Ser Gly Cys Thr 
305                 310                 315                 320 


Arg Glu Asn Leu Glu Ala Leu Ile Lys Glu Phe Leu Asp Tyr Leu Glu 
                325                 330                 335     


Ile Asp Phe Glu Ile Val Gly Asp Ser Cys Met Val Tyr Gly Asp Thr 
            340                 345                 350         


Leu Asp Ile Met His Gly Asp Leu Glu Leu Ser Ser Ala Val Val Gly 
        355                 360                 365             


Pro Val Ser Leu Asp Arg Glu Trp Gly Ile Asp Lys Pro Thr Ile Gly 
    370                 375                 380                 


Ala Gly Phe Gly Leu Glu Arg Leu Leu Lys Val Met His Gly Phe Lys 
385                 390                 395                 400 


Asn Ile Lys Arg Ala Ser Arg Ser Glu Ser Tyr Tyr Asn Gly Ile Ser 
                405                 410                 415     


Thr Asn Leu 
            


