               SEQUENCE LISTING

<110> Technische Universität Hamburg

<120> NOVEL DHAP SYNTHASE

<130> PAT 1716 WO

<150> LU101172
<151> 2019-04-10
<150> DE102019109423.2
<151> 2019-04-10

<160> 35

<170> BiSSAP 1.3.6

<210> 1
<211> 350
<212> PRT
<213> Artificial Sequence


<220> 
<223> AroG mutant general sequence

<220> 
<221> SITE
<222> 6..7
<223> Xaa = any amino acid, not both Asp

<400> 1
Met Asn Tyr Gln Asn Xaa Xaa Leu Arg Ile Lys Glu Ile Lys Glu Leu 
1               5                   10                  15      
Leu Pro Pro Val Ala Leu Leu Glu Lys Phe Pro Ala Thr Glu Asn Ala 
            20                  25                  30          
Ala Asn Thr Val Ala His Ala Arg Lys Ala Ile His Lys Ile Leu Lys 
        35                  40                  45              
Gly Asn Asp Asp Arg Leu Leu Val Val Ile Gly Pro Cys Ser Ile His 
    50                  55                  60                  
Asp Pro Val Ala Ala Lys Glu Tyr Ala Thr Arg Leu Leu Ala Leu Arg 
65                  70                  75                  80  
Glu Glu Leu Lys Asp Glu Leu Glu Ile Val Met Arg Val Tyr Phe Glu 
                85                  90                  95      
Lys Pro Arg Thr Thr Val Gly Trp Lys Gly Leu Ile Asn Asp Pro His 
            100                 105                 110         
Met Asp Asn Ser Phe Gln Ile Asn Asp Gly Leu Arg Ile Ala Arg Lys 
        115                 120                 125             
Leu Leu Leu Asp Ile Asn Asp Ser Gly Leu Pro Ala Ala Gly Glu Phe 
    130                 135                 140                 
Leu Asp Met Ile Thr Pro Gln Tyr Leu Ala Asp Leu Met Ser Trp Gly 
145                 150                 155                 160 
Ala Ile Gly Ala Arg Thr Thr Glu Ser Gln Val His Arg Glu Leu Ala 
                165                 170                 175     
Ser Gly Leu Ser Cys Pro Val Gly Phe Lys Asn Gly Thr Asp Gly Thr 
            180                 185                 190         
Ile Lys Val Ala Ile Asp Ala Ile Asn Ala Ala Gly Ala Pro His Cys 
        195                 200                 205             
Phe Leu Ser Val Thr Lys Trp Gly His Ser Ala Ile Val Asn Thr Ser 
    210                 215                 220                 
Gly Asn Gly Asp Cys His Ile Ile Leu Arg Gly Gly Lys Glu Pro Asn 
225                 230                 235                 240 
Tyr Ser Ala Lys His Val Ala Glu Val Lys Glu Gly Leu Asn Lys Ala 
                245                 250                 255     
Gly Leu Pro Ala Gln Val Met Ile Asp Phe Ser His Ala Asn Ser Ser 
            260                 265                 270         
Lys Gln Phe Lys Lys Gln Met Asp Val Cys Ala Asp Val Cys Gln Gln 
        275                 280                 285             
Ile Ala Gly Gly Glu Lys Ala Ile Ile Gly Val Met Val Glu Ser His 
    290                 295                 300                 
Leu Val Glu Gly Asn Gln Ser Leu Glu Ser Gly Glu Pro Leu Ala Tyr 
305                 310                 315                 320 
Gly Lys Ser Ile Thr Asp Ala Cys Ile Gly Trp Glu Asp Thr Asp Ala 
                325                 330                 335     
Leu Leu Arg Gln Leu Ala Asn Ala Val Lys Ala Arg Arg Gly 
            340                 345                 350 

<210> 2
<211> 350
<212> PRT
<213> Artificial Sequence


<220> 
<223> AroG mutant D6G-D7A

<400> 2
Met Asn Tyr Gln Asn Gly Ala Leu Arg Ile Lys Glu Ile Lys Glu Leu 
1               5                   10                  15      
Leu Pro Pro Val Ala Leu Leu Glu Lys Phe Pro Ala Thr Glu Asn Ala 
            20                  25                  30          
Ala Asn Thr Val Ala His Ala Arg Lys Ala Ile His Lys Ile Leu Lys 
        35                  40                  45              
Gly Asn Asp Asp Arg Leu Leu Val Val Ile Gly Pro Cys Ser Ile His 
    50                  55                  60                  
Asp Pro Val Ala Ala Lys Glu Tyr Ala Thr Arg Leu Leu Ala Leu Arg 
65                  70                  75                  80  
Glu Glu Leu Lys Asp Glu Leu Glu Ile Val Met Arg Val Tyr Phe Glu 
                85                  90                  95      
Lys Pro Arg Thr Thr Val Gly Trp Lys Gly Leu Ile Asn Asp Pro His 
            100                 105                 110         
Met Asp Asn Ser Phe Gln Ile Asn Asp Gly Leu Arg Ile Ala Arg Lys 
        115                 120                 125             
Leu Leu Leu Asp Ile Asn Asp Ser Gly Leu Pro Ala Ala Gly Glu Phe 
    130                 135                 140                 
Leu Asp Met Ile Thr Pro Gln Tyr Leu Ala Asp Leu Met Ser Trp Gly 
145                 150                 155                 160 
Ala Ile Gly Ala Arg Thr Thr Glu Ser Gln Val His Arg Glu Leu Ala 
                165                 170                 175     
Ser Gly Leu Ser Cys Pro Val Gly Phe Lys Asn Gly Thr Asp Gly Thr 
            180                 185                 190         
Ile Lys Val Ala Ile Asp Ala Ile Asn Ala Ala Gly Ala Pro His Cys 
        195                 200                 205             
Phe Leu Ser Val Thr Lys Trp Gly His Ser Ala Ile Val Asn Thr Ser 
    210                 215                 220                 
Gly Asn Gly Asp Cys His Ile Ile Leu Arg Gly Gly Lys Glu Pro Asn 
225                 230                 235                 240 
Tyr Ser Ala Lys His Val Ala Glu Val Lys Glu Gly Leu Asn Lys Ala 
                245                 250                 255     
Gly Leu Pro Ala Gln Val Met Ile Asp Phe Ser His Ala Asn Ser Ser 
            260                 265                 270         
Lys Gln Phe Lys Lys Gln Met Asp Val Cys Ala Asp Val Cys Gln Gln 
        275                 280                 285             
Ile Ala Gly Gly Glu Lys Ala Ile Ile Gly Val Met Val Glu Ser His 
    290                 295                 300                 
Leu Val Glu Gly Asn Gln Ser Leu Glu Ser Gly Glu Pro Leu Ala Tyr 
305                 310                 315                 320 
Gly Lys Ser Ile Thr Asp Ala Cys Ile Gly Trp Glu Asp Thr Asp Ala 
                325                 330                 335     
Leu Leu Arg Gln Leu Ala Asn Ala Val Lys Ala Arg Arg Gly 
            340                 345                 350 

<210> 3
<211> 350
<212> PRT
<213> Artificial Sequence


<220> 
<223> AroG mutant D6L-D7P

<400> 3
Met Asn Tyr Gln Asn Leu Pro Leu Arg Ile Lys Glu Ile Lys Glu Leu 
1               5                   10                  15      
Leu Pro Pro Val Ala Leu Leu Glu Lys Phe Pro Ala Thr Glu Asn Ala 
            20                  25                  30          
Ala Asn Thr Val Ala His Ala Arg Lys Ala Ile His Lys Ile Leu Lys 
        35                  40                  45              
Gly Asn Asp Asp Arg Leu Leu Val Val Ile Gly Pro Cys Ser Ile His 
    50                  55                  60                  
Asp Pro Val Ala Ala Lys Glu Tyr Ala Thr Arg Leu Leu Ala Leu Arg 
65                  70                  75                  80  
Glu Glu Leu Lys Asp Glu Leu Glu Ile Val Met Arg Val Tyr Phe Glu 
                85                  90                  95      
Lys Pro Arg Thr Thr Val Gly Trp Lys Gly Leu Ile Asn Asp Pro His 
            100                 105                 110         
Met Asp Asn Ser Phe Gln Ile Asn Asp Gly Leu Arg Ile Ala Arg Lys 
        115                 120                 125             
Leu Leu Leu Asp Ile Asn Asp Ser Gly Leu Pro Ala Ala Gly Glu Phe 
    130                 135                 140                 
Leu Asp Met Ile Thr Pro Gln Tyr Leu Ala Asp Leu Met Ser Trp Gly 
145                 150                 155                 160 
Ala Ile Gly Ala Arg Thr Thr Glu Ser Gln Val His Arg Glu Leu Ala 
                165                 170                 175     
Ser Gly Leu Ser Cys Pro Val Gly Phe Lys Asn Gly Thr Asp Gly Thr 
            180                 185                 190         
Ile Lys Val Ala Ile Asp Ala Ile Asn Ala Ala Gly Ala Pro His Cys 
        195                 200                 205             
Phe Leu Ser Val Thr Lys Trp Gly His Ser Ala Ile Val Asn Thr Ser 
    210                 215                 220                 
Gly Asn Gly Asp Cys His Ile Ile Leu Arg Gly Gly Lys Glu Pro Asn 
225                 230                 235                 240 
Tyr Ser Ala Lys His Val Ala Glu Val Lys Glu Gly Leu Asn Lys Ala 
                245                 250                 255     
Gly Leu Pro Ala Gln Val Met Ile Asp Phe Ser His Ala Asn Ser Ser 
            260                 265                 270         
Lys Gln Phe Lys Lys Gln Met Asp Val Cys Ala Asp Val Cys Gln Gln 
        275                 280                 285             
Ile Ala Gly Gly Glu Lys Ala Ile Ile Gly Val Met Val Glu Ser His 
    290                 295                 300                 
Leu Val Glu Gly Asn Gln Ser Leu Glu Ser Gly Glu Pro Leu Ala Tyr 
305                 310                 315                 320 
Gly Lys Ser Ile Thr Asp Ala Cys Ile Gly Trp Glu Asp Thr Asp Ala 
                325                 330                 335     
Leu Leu Arg Gln Leu Ala Asn Ala Val Lys Ala Arg Arg Gly 
            340                 345                 350 

<210> 4
<211> 350
<212> PRT
<213> Artificial Sequence


<220> 
<223> AroG mutant D6P-D7I

<400> 4
Met Asn Tyr Gln Asn Pro Ile Leu Arg Ile Lys Glu Ile Lys Glu Leu 
1               5                   10                  15      
Leu Pro Pro Val Ala Leu Leu Glu Lys Phe Pro Ala Thr Glu Asn Ala 
            20                  25                  30          
Ala Asn Thr Val Ala His Ala Arg Lys Ala Ile His Lys Ile Leu Lys 
        35                  40                  45              
Gly Asn Asp Asp Arg Leu Leu Val Val Ile Gly Pro Cys Ser Ile His 
    50                  55                  60                  
Asp Pro Val Ala Ala Lys Glu Tyr Ala Thr Arg Leu Leu Ala Leu Arg 
65                  70                  75                  80  
Glu Glu Leu Lys Asp Glu Leu Glu Ile Val Met Arg Val Tyr Phe Glu 
                85                  90                  95      
Lys Pro Arg Thr Thr Val Gly Trp Lys Gly Leu Ile Asn Asp Pro His 
            100                 105                 110         
Met Asp Asn Ser Phe Gln Ile Asn Asp Gly Leu Arg Ile Ala Arg Lys 
        115                 120                 125             
Leu Leu Leu Asp Ile Asn Asp Ser Gly Leu Pro Ala Ala Gly Glu Phe 
    130                 135                 140                 
Leu Asp Met Ile Thr Pro Gln Tyr Leu Ala Asp Leu Met Ser Trp Gly 
145                 150                 155                 160 
Ala Ile Gly Ala Arg Thr Thr Glu Ser Gln Val His Arg Glu Leu Ala 
                165                 170                 175     
Ser Gly Leu Ser Cys Pro Val Gly Phe Lys Asn Gly Thr Asp Gly Thr 
            180                 185                 190         
Ile Lys Val Ala Ile Asp Ala Ile Asn Ala Ala Gly Ala Pro His Cys 
        195                 200                 205             
Phe Leu Ser Val Thr Lys Trp Gly His Ser Ala Ile Val Asn Thr Ser 
    210                 215                 220                 
Gly Asn Gly Asp Cys His Ile Ile Leu Arg Gly Gly Lys Glu Pro Asn 
225                 230                 235                 240 
Tyr Ser Ala Lys His Val Ala Glu Val Lys Glu Gly Leu Asn Lys Ala 
                245                 250                 255     
Gly Leu Pro Ala Gln Val Met Ile Asp Phe Ser His Ala Asn Ser Ser 
            260                 265                 270         
Lys Gln Phe Lys Lys Gln Met Asp Val Cys Ala Asp Val Cys Gln Gln 
        275                 280                 285             
Ile Ala Gly Gly Glu Lys Ala Ile Ile Gly Val Met Val Glu Ser His 
    290                 295                 300                 
Leu Val Glu Gly Asn Gln Ser Leu Glu Ser Gly Glu Pro Leu Ala Tyr 
305                 310                 315                 320 
Gly Lys Ser Ile Thr Asp Ala Cys Ile Gly Trp Glu Asp Thr Asp Ala 
                325                 330                 335     
Leu Leu Arg Gln Leu Ala Asn Ala Val Lys Ala Arg Arg Gly 
            340                 345                 350 

<210> 5
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer pTargR

<400> 5
actagtatta tacctaggac tgagctagct gtcaag                               36


<210> 6
<211> 87
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer AroG-N20

<400> 6
gctcagtcct aggtataata ctagtcagga agcagtgcgg cgcacgtttt agagctagaa     60

atagcaagtt aaaataaggc tagtccg                                         87


<210> 7
<211> 87
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer TrpR-N20

<400> 7
gctcagtcct aggtataata ctagttcagg tttaacaacg gtaaagtttt agagctagaa     60

atagcaagtt aaaataaggc tagtccg                                         87


<210> 8
<211> 87
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Cm-N20

<400> 8
gctcagtcct aggtataata ctagttgatg aacctgaatc gccaggtttt agagctagaa     60

atagcaagtt aaaataaggc tagtccg                                         87


<210> 9
<211> 87
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Cm-N202

<400> 9
gctcagtcct aggtataata ctagtgctga tgccgctggc gattcgtttt agagctagaa     60

atagcaagtt aaaataaggc tagtccg                                         87


<210> 10
<211> 26
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer serA-Fus-F

<400> 10
cctacggtaa gagcatcacc gatgcc                                         26


<210> 11
<211> 25
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer serA-Fus-R

<400> 11
gcccaattgc gtaccaatat gaccg                                          25


<210> 12
<211> 30
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Cm-C1

<400> 12
gatataccac cgttgatata tcccaatggc                                     30


<210> 13
<211> 25
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Cm-C2

<400> 13
ttacatcagc accttgtcgc cttgc                                          25


<210> 14
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer pN20VRCm

<400> 14
caacggtggt atataaaaaa gcaccgactc ggtgcc                               36


<210> 15
<211> 39
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer pN20VFSerA

<400> 15
ggtacgcaat tgggctcgag gtgaagacga aagggcctc                            39


<210> 16
<211> 53
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer WaroG-FF

<400> 16
ggcgacaagg tgctgatgta atattgcatt cactaagata agtatggcaa cac            53


<210> 17
<211> 26
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer aroG-fus-R

<400> 17
ggcatcggtg atgctcttac cgtagg                                         26


<210> 18
<211> 71
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer D67X-F

<220> 
<221> misc_feature
<222> 25..26
<223> /note="n = A, C, G or T"

<220> 
<221> misc_feature
<222> 28..29
<223> /note="n = A, C, G or T"

<400> 18
ggaacagaca tgaattatca gaacnnknnk ttacgcatca aagaaatcaa agagttactt     60

cctcctgtcg c                                                          71


<210> 19
<211> 33
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer D67X-R

<400> 19
gttctgataa ttcatgtctg ttccagtgtt gcc                                  33


<210> 20
<211> 30
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer aroG_S180F_F

<400> 20
gcatcagggc ttttttgtcc ggtcggcttc                                     30


<210> 21
<211> 30
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer aroG_S180F_R

<400> 21
gaagccgacc ggacaaaaaa gccctgatgc                                     30


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


<220> 
<223> Primer pTagC1

<400> 22
ttgagtgagc tgataccgct cgc                                            23


<210> 23
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer XbaI-serA

<400> 23
gagcggataa caattcccct c                                              21


<210> 24
<211> 62
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer aroG-His-HindIII

<400> 24
cgccggaagc tttcattagt ggtggtggtg gtggtggccc gcgacgcgct tttactgcat     60

tc                                                                    62


<210> 25
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer D67M-F

<400> 25
ttacgcatca aagaaatcaa agacttactt cc                                   32


<210> 26
<211> 57
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer D67M-LPR

<400> 26
ctctttgatt tctttgatgc gtaaaggcaa gttctgataa ttcatatgta tatctcc        57


<210> 27
<211> 57
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer D67M-PIR

<400> 27
ctctttgatt tctttgatgc gtaaaatagg gttctgataa ttcatatgta tatctcc        57


<210> 28
<211> 57
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer D67M-GAR

<400> 28
ctctttgatt tctttgatgc gtaaagcccc gttctgataa ttcatatgta tatctcc        57


<210> 29
<211> 84
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Tsen-trpR-IF

<400> 29
ggataaaccg acgttgatga gcgccacgga atggggacgt cgttactgat ccgcacggct     60

gttgacaatt aatcatccgg ctcg                                            84


<210> 30
<211> 87
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Tsen-trpR-IR

<400> 30
tcaatcgctt ttcagcaaca cctcttccag ccactggcgc agctcgacgg gcgcggcttt     60

cttttacttg tacagctcgt ccatgcc                                         87


<210> 31
<211> 80
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer up-aroH-out

<400> 31
ggggcgttgg tgtaaagatt attgccctca ccctgtacgg gtgagggcgt agagagatta     60

cgcggccgct tctagagttg                                                 80


<210> 32
<211> 72
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer Cm-delG-R

<400> 32
gtatcttccc agcctatgca ggcatcggtg atgctcttac cgtaggccag cacctgaagt     60

cagccccata cg                                                         72


<210> 33
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer NotI-pTac-aroG

<400> 33
agtgcagcgg ccgctgttga caattaatca tcggctcgta taatgtgtag gggaattgtg     60

agcggataac                                                            70


<210> 34
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer aroG-speI

<400> 34
tgcggcacta gtttattacc cgcgacgcgc ttttac                               36


<210> 35
<211> 85
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer u-rpsLp-tac

<400> 35
ttgtgtgagg acgttttatt acgtgtttac gaagcaaaag ctaaaaccag gagctattta     60

ctgttgacaa ttaatcatcg gctcg                                           85


