                         SEQUENCE LISTING

<110>  Roche Diagnostics GmbH
       F. Hoffmann-La Roche AG
       Roche Molecular Systems, Inc.
 
<120>  IMPROVED MODIFIED/MUTANT BACTERIAL LUCIFERASES

<130>  P34374-WO-HS

<150>  62/576,814
<151>  2017-10-25

<160>  3     

<170>  PatentIn version 3.5

<210>  1
<211>  2092
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  E. coli codon-optimized wild type LuxAB (Aliivibrio fischeri)

<400>  1
atgaaatttg gtaacatctg cttctcgtac caaccgcctg gcgaaactca caaacaagtg       60

atggaccgtt tcgttcgcct gggtattgcg agcgaagagg tcggcttcga cacctactgg      120

accttggagc atcattttac cgagtttggt ttgaccggca acctgtttgt ggcggcagca      180

aatctgttgg gtcgtaccaa gacgctgaat gttggcacca tgggcgttgt gattccgacg      240

gcacacccgg tgcgccagct ggaagatgtg ctgctgctgg atcagatgtc caagggccgc      300

ttcaacttcg gtaccgtgcg cggtctgtac cacaaagatt tccgtgtttt tggcgtcgac      360

atggaagaga gccgtgccat tacgcaaaac ttttatcaaa tgattatgga aagcctgcag      420

accggtacca tcagcagcga ttctgattac attcagtttc cgaaagtcga cgtctatcca      480

aaggtttatt ccaaaaacgt tccgacgtgt atgaccgcag agtctgcgag cacgacggaa      540

tggctggcta ttcagggtct gccgatggtc ttatcctgga ttatcggtac gaatgagaaa      600

aaagcgcaaa tggaattgta caacgagatt gccaccgagt atggccatga catcagcaag      660

atcgaccact gtatgaccta tatctgcagc gttgacgatg acgcgcaaaa ggctcaagac      720

gtttgccgtg agtttctgaa gaactggtat gattcttacg tgaatgcgac gaacattttc      780

aatgatagca atcagacccg tggttacgac taccacaagg gtcagtggcg cgatttcgtt      840

ctgcagggtc atacgaatac taatcgtcgt gtcgattaca gcaacggtat taacccggta      900

ggcaccccgg agcaatgcat cgagatcatc cagcgtgaca tcgacgctac cggtatcacg      960

aatatcacgt gtggttttga ggcgaacggt actgaagatg aaatcatcgc cagcatgcgt     1020

cgcttcatga cccaagtggc gccgttcctg aaagaaccga agtgaggatc cctaggaata     1080

attttgttta actttaagaa ggagataaaa aatgaaattt ggtctgtttt tcctgaattt     1140

tcaaaaggat ggcatcacga gcgaagaaac cctggacaac atggtcaaga ctgttacgtt     1200

gattgatagc accaagtatc acttcaatac ggcctttgtt aacgaacacc actttagcaa     1260

aaacggtatt gtgggcgcac cgattaccgc ggctggtttt ctgctgggtc tgaccaacaa     1320

gctccacatc ggctcgctga accaagtcat caccacccac cacccggttc gtgtcgcaga     1380

agaggcgagc ctgttggacc agatgtctga gggtcgtttc attctgggtt tcagcgattg     1440

tgagagcgac ttcgagatgg aatttttccg tcgtcatatc agcagccgtc aacaacagtt     1500

tgaggcatgc tatgaaatca ttaatgacgc cctgaccacc ggctactgtc atccgcagaa     1560

tgatttctac gatttcccta aggttagcat taacccgcat tgctattccg agaatggccc     1620

aaagcaatat gtgagcgcaa ccagcaaaga agtcgtgatg tgggctgcga agaaagcgct     1680

gccgctgacg ttcaaatggg aagataacct ggaaacgaaa gagcgctacg cgatcctgta     1740

taacaagacc gcgcagcagt acggtatcga catctctgac gtggaccacc agttgaccgt     1800

tattgcgaat ctgaatgccg atcgcagcac ggcgcaggaa gaggttcgcg agtacctgaa     1860

agattacatt acggaaacct acccgcagat ggaccgtgac gagaaaatca actgcatcat     1920

tgaagagaac gcagtgggct cccatgacga ctactatgag agcactaaac tggccgtcga     1980

gaaaaccggt tccaagaaca ttctgttgag cttcgagagc atgagcgata tcaaagatgt     2040

gaaagacatt atcgacatgc tgaatcaaaa gattgagatg aatctgccgt ga             2092


<210>  2
<211>  354
<212>  PRT
<213>  Aliivibrio fischeri (LuxA)

<400>  2

Met Lys Phe Gly Asn Ile Cys Phe Ser Tyr Gln Pro Pro Gly Glu Thr 
1               5                   10                  15      


His Lys Gln Val Met Asp Arg Phe Val Arg Leu Gly Ile Ala Ser Glu 
            20                  25                  30          


Glu Val Gly Phe Asp Thr Tyr Trp Thr Leu Glu His His Phe Thr Glu 
        35                  40                  45              


Phe Gly Leu Thr Gly Asn Leu Phe Val Ala Ala Ala Asn Leu Leu Gly 
    50                  55                  60                  


Arg Thr Lys Thr Leu Asn Val Gly Thr Met Gly Val Val Ile Pro Thr 
65                  70                  75                  80  


Ala His Pro Val Arg Gln Leu Glu Asp Val Leu Leu Leu Asp Gln Met 
                85                  90                  95      


Ser Lys Gly Arg Phe Asn Phe Gly Thr Val Arg Gly Leu Tyr His Lys 
            100                 105                 110         


Asp Phe Arg Val Phe Gly Val Asp Met Glu Glu Ser Arg Ala Ile Thr 
        115                 120                 125             


Gln Asn Phe Tyr Gln Met Ile Met Glu Ser Leu Gln Thr Gly Thr Ile 
    130                 135                 140                 


Ser Ser Asp Ser Asp Tyr Ile Gln Phe Pro Lys Val Asp Val Tyr Pro 
145                 150                 155                 160 


Lys Val Tyr Ser Lys Asn Val Pro Thr Cys Met Thr Ala Glu Ser Ala 
                165                 170                 175     


Ser Thr Thr Glu Trp Leu Ala Ile Gln Gly Leu Pro Met Val Leu Ser 
            180                 185                 190         


Trp Ile Ile Gly Thr Asn Glu Lys Lys Ala Gln Met Glu Leu Tyr Asn 
        195                 200                 205             


Glu Ile Ala Thr Glu Tyr Gly His Asp Ile Ser Lys Ile Asp His Cys 
    210                 215                 220                 


Met Thr Tyr Ile Cys Ser Val Asp Asp Asp Ala Gln Lys Ala Gln Asp 
225                 230                 235                 240 


Val Cys Arg Glu Phe Leu Lys Asn Trp Tyr Asp Ser Tyr Val Asn Ala 
                245                 250                 255     


Thr Asn Ile Phe Asn Asp Ser Asn Gln Thr Arg Gly Tyr Asp Tyr His 
            260                 265                 270         


Lys Gly Gln Trp Arg Asp Phe Val Leu Gln Gly His Thr Asn Thr Asn 
        275                 280                 285             


Arg Arg Val Asp Tyr Ser Asn Gly Ile Asn Pro Val Gly Thr Pro Glu 
    290                 295                 300                 


Gln Cys Ile Glu Ile Ile Gln Arg Asp Ile Asp Ala Thr Gly Ile Thr 
305                 310                 315                 320 


Asn Ile Thr Cys Gly Phe Glu Ala Asn Gly Thr Glu Asp Glu Ile Ile 
                325                 330                 335     


Ala Ser Met Arg Arg Phe Met Thr Gln Val Ala Pro Phe Leu Lys Glu 
            340                 345                 350         


Pro Lys 
        


<210>  3
<211>  326
<212>  PRT
<213>  Aliivibrio fischeri (LuxB)

<400>  3

Met Lys Phe Gly Leu Phe Phe Leu Asn Phe Gln Lys Asp Gly Ile Thr 
1               5                   10                  15      


Ser Glu Glu Thr Leu Asp Asn Met Val Lys Thr Val Thr Leu Ile Asp 
            20                  25                  30          


Ser Thr Lys Tyr His Phe Asn Thr Ala Phe Val Asn Glu His His Phe 
        35                  40                  45              


Ser Lys Asn Gly Ile Val Gly Ala Pro Ile Thr Ala Ala Gly Phe Leu 
    50                  55                  60                  


Leu Gly Leu Thr Asn Lys Leu His Ile Gly Ser Leu Asn Gln Val Ile 
65                  70                  75                  80  


Thr Thr His His Pro Val Arg Val Ala Glu Glu Ala Ser Leu Leu Asp 
                85                  90                  95      


Gln Met Ser Glu Gly Arg Phe Ile Leu Gly Phe Ser Asp Cys Glu Ser 
            100                 105                 110         


Asp Phe Glu Met Glu Phe Phe Arg Arg His Ile Ser Ser Arg Gln Gln 
        115                 120                 125             


Gln Phe Glu Ala Cys Tyr Glu Ile Ile Asn Asp Ala Leu Thr Thr Gly 
    130                 135                 140                 


Tyr Cys His Pro Gln Asn Asp Phe Tyr Asp Phe Pro Lys Val Ser Ile 
145                 150                 155                 160 


Asn Pro His Cys Tyr Ser Glu Asn Gly Pro Lys Gln Tyr Val Ser Ala 
                165                 170                 175     


Thr Ser Lys Glu Val Val Met Trp Ala Ala Lys Lys Ala Leu Pro Leu 
            180                 185                 190         


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


Leu Tyr Asn Lys Thr Ala Gln Gln Tyr Gly Ile Asp Ile Ser Asp Val 
    210                 215                 220                 


Asp His Gln Leu Thr Val Ile Ala Asn Leu Asn Ala Asp Arg Ser Thr 
225                 230                 235                 240 


Ala Gln Glu Glu Val Arg Glu Tyr Leu Lys Asp Tyr Ile Thr Glu Thr 
                245                 250                 255     


Tyr Pro Gln Met Asp Arg Asp Glu Lys Ile Asn Cys Ile Ile Glu Glu 
            260                 265                 270         


Asn Ala Val Gly Ser His Asp Asp Tyr Tyr Glu Ser Thr Lys Leu Ala 
        275                 280                 285             


Val Glu Lys Thr Gly Ser Lys Asn Ile Leu Leu Ser Phe Glu Ser Met 
    290                 295                 300                 


Ser Asp Ile Lys Asp Val Lys Asp Ile Ile Asp Met Leu Asn Gln Lys 
305                 310                 315                 320 


Ile Glu Met Asn Leu Pro 
                325     


