                         SEQUENCE LISTING

<110>  Scarab Genomics, LLC
 
<120>  METHODS AND COMPOSITIONS FOR IMPROVED EXPRESSION OF RECOMBINANT 
       PROTEINS

<130>  010447-5043 WO

<140>  PCT/US18/49422
<141>  2018-09-04

<150>  US 62/554,443
<151>  2017-09-05

<160>  9     

<170>  PatentIn version 3.5

<210>  1
<211>  358
<212>  PRT
<213>  Escherichia coli

<400>  1

Met Lys Leu Thr Leu Lys Asn Leu Ser Met Ala Ile Met Met Ser Thr 
1               5                   10                  15      


Ile Val Met Gly Ser Ser Ala Met Ala Ala Asp Ser Asn Glu Lys Ile 
            20                  25                  30          


Val Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro Glu His Thr Leu 
        35                  40                  45              


Pro Ala Lys Ala Met Ala Tyr Ala Gln Gly Ala Asp Tyr Leu Glu Gln 
    50                  55                  60                  


Asp Leu Val Met Thr Lys Asp Asp Asn Leu Val Val Leu His Asp His 
65                  70                  75                  80  


Tyr Leu Asp Arg Val Thr Asp Val Ala Asp Arg Phe Pro Asp Arg Ala 
                85                  90                  95      


Arg Lys Asp Gly Arg Tyr Tyr Ala Ile Asp Phe Thr Leu Asp Glu Ile 
            100                 105                 110         


Lys Ser Leu Lys Phe Thr Glu Gly Phe Asp Ile Glu Asn Gly Lys Lys 
        115                 120                 125             


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


Val His Thr Phe Glu Glu Glu Ile Glu Phe Val Gln Gly Leu Asn His 
145                 150                 155                 160 


Ser Thr Gly Lys Asn Ile Gly Ile Tyr Pro Glu Ile Lys Ala Pro Trp 
                165                 170                 175     


Phe His His Gln Glu Gly Lys Asp Ile Ala Ala Lys Thr Leu Glu Val 
            180                 185                 190         


Leu Lys Lys Tyr Gly Tyr Thr Gly Lys Asp Asp Lys Val Tyr Leu Gln 
        195                 200                 205             


Cys Phe Asp Ala Asp Glu Leu Lys Arg Ile Lys Asn Glu Leu Glu Pro 
    210                 215                 220                 


Lys Met Gly Met Glu Leu Asn Leu Val Gln Leu Ile Ala Tyr Thr Asp 
225                 230                 235                 240 


Trp Asn Glu Thr Gln Gln Lys Gln Pro Asp Gly Ser Trp Val Asn Tyr 
                245                 250                 255     


Asn Tyr Asp Trp Met Phe Lys Pro Gly Ala Met Lys Gln Val Ala Glu 
            260                 265                 270         


Tyr Ala Asp Gly Ile Gly Pro Asp Tyr His Met Leu Ile Glu Glu Thr 
        275                 280                 285             


Ser Gln Pro Gly Asn Ile Lys Leu Thr Gly Met Val Gln Asp Ala Gln 
    290                 295                 300                 


Gln Asn Lys Leu Val Val His Pro Tyr Thr Val Arg Ser Asp Lys Leu 
305                 310                 315                 320 


Pro Glu Tyr Thr Pro Asp Val Asn Gln Leu Tyr Asp Ala Leu Tyr Asn 
                325                 330                 335     


Lys Ala Gly Val Asn Gly Leu Phe Thr Asp Phe Pro Asp Lys Ala Val 
            340                 345                 350         


Lys Phe Leu Asn Lys Glu 
        355             


<210>  2
<211>  319
<212>  PRT
<213>  Escherichia coli

<400>  2

Met Lys Pro Glu Asn Lys Leu Pro Val Leu Asp Leu Ile Ser Ala Glu 
1               5                   10                  15      


Met Lys Thr Val Val Asn Thr Leu Gln Pro Asp Leu Pro Pro Trp Pro 
            20                  25                  30          


Ala Thr Gly Thr Ile Ala Glu Gln Arg Gln Tyr Tyr Thr Leu Glu Arg 
        35                  40                  45              


Arg Phe Trp Asn Ala Gly Ala Pro Glu Met Ala Thr Arg Ala Tyr Met 
    50                  55                  60                  


Val Pro Thr Lys Tyr Gly Gln Val Glu Thr Arg Leu Phe Cys Pro Gln 
65                  70                  75                  80  


Pro Asp Ser Pro Ala Thr Leu Phe Tyr Leu His Gly Gly Gly Phe Ile 
                85                  90                  95      


Leu Gly Asn Leu Asp Thr His Asp Arg Ile Met Arg Leu Leu Ala Ser 
            100                 105                 110         


Tyr Ser Gln Cys Thr Val Ile Gly Ile Asp Tyr Thr Leu Ser Pro Glu 
        115                 120                 125             


Ala Arg Phe Pro Gln Ala Ile Glu Glu Ile Val Ala Ala Cys Cys Tyr 
    130                 135                 140                 


Phe His Gln Gln Ala Glu Asp Tyr Gln Ile Asn Met Ser Arg Ile Gly 
145                 150                 155                 160 


Phe Ala Gly Asp Ser Ala Gly Ala Met Leu Ala Leu Ala Ser Ala Leu 
                165                 170                 175     


Trp Leu Arg Asp Lys Gln Ile Asp Cys Gly Lys Val Ala Gly Val Leu 
            180                 185                 190         


Leu Trp Tyr Gly Leu Tyr Gly Leu Arg Asp Ser Val Thr Arg Arg Leu 
        195                 200                 205             


Leu Gly Gly Val Trp Asp Gly Leu Thr Gln Gln Asp Leu Gln Met Tyr 
    210                 215                 220                 


Glu Glu Ala Tyr Leu Ser Asn Asp Ala Asp Arg Glu Ser Pro Tyr Tyr 
225                 230                 235                 240 


Cys Leu Phe Asn Asn Asp Leu Thr Arg Glu Val Pro Pro Cys Phe Ile 
                245                 250                 255     


Ala Gly Ala Glu Phe Asp Pro Leu Leu Asp Asp Ser Arg Leu Leu Tyr 
            260                 265                 270         


Gln Thr Leu Ala Ala His Gln Gln Pro Cys Glu Phe Lys Leu Tyr Pro 
        275                 280                 285             


Gly Thr Leu His Ala Phe Leu His Tyr Ser Arg Met Met Lys Thr Ala 
    290                 295                 300                 


Asp Glu Ala Leu Arg Asp Gly Ala Gln Phe Phe Thr Ala Gln Leu 
305                 310                 315                 


<210>  3
<211>  340
<212>  PRT
<213>  Escherichia coli

<400>  3

Met Phe Gln Gln Gln Lys Asp Trp Glu Thr Arg Glu Asn Ala Phe Ala 
1               5                   10                  15      


Ala Phe Thr Met Gly Pro Leu Thr Asp Phe Trp Arg Gln Arg Asp Glu 
            20                  25                  30          


Ala Glu Phe Thr Gly Val Asp Asp Ile Pro Val Arg Phe Val Arg Phe 
        35                  40                  45              


Arg Ala Gln His His Asp Arg Val Val Val Ile Cys Pro Gly Arg Ile 
    50                  55                  60                  


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


Gly Phe Asp Val Leu Ile Ile Asp His Arg Gly Gln Gly Arg Ser Gly 
                85                  90                  95      


Arg Leu Leu Ala Asp Pro His Leu Gly His Val Asn Arg Phe Asn Asp 
            100                 105                 110         


Tyr Val Asp Asp Leu Ala Ala Phe Trp Gln Gln Glu Val Gln Pro Gly 
        115                 120                 125             


Pro Trp Arg Lys Arg Tyr Ile Leu Ala His Ser Met Gly Gly Ala Ile 
    130                 135                 140                 


Ser Thr Leu Phe Leu Gln Arg His Pro Gly Val Cys Asp Ala Ile Ala 
145                 150                 155                 160 


Leu Thr Ala Pro Met Phe Gly Ile Val Ile Arg Met Pro Ser Phe Met 
                165                 170                 175     


Ala Arg Gln Ile Leu Asn Trp Ala Glu Ala His Pro Arg Phe Arg Asp 
            180                 185                 190         


Gly Tyr Ala Ile Gly Thr Gly Arg Trp Arg Ala Leu Pro Phe Ala Ile 
        195                 200                 205             


Asn Val Leu Thr His Ser Arg Gln Arg Tyr Arg Arg Asn Leu Arg Phe 
    210                 215                 220                 


Tyr Ala Asp Asp Pro Thr Ile Arg Val Gly Gly Pro Thr Tyr His Trp 
225                 230                 235                 240 


Val Arg Glu Ser Ile Leu Ala Gly Glu Gln Val Leu Ala Gly Ala Gly 
                245                 250                 255     


Asp Asp Ala Thr Pro Thr Leu Leu Leu Gln Ala Glu Glu Glu Arg Val 
            260                 265                 270         


Val Asp Asn Arg Met His Asp Arg Phe Cys Glu Leu Arg Thr Ala Ala 
        275                 280                 285             


Gly His Pro Val Glu Gly Gly Arg Pro Leu Val Ile Lys Gly Ala Tyr 
    290                 295                 300                 


His Glu Ile Leu Phe Glu Lys Asp Ala Met Arg Ser Val Ala Leu His 
305                 310                 315                 320 


Ala Ile Val Asp Phe Phe Asn Arg His Asn Ser Pro Ser Gly Asn Arg 
                325                 330                 335     


Ser Thr Glu Val 
            340 


<210>  4
<211>  289
<212>  PRT
<213>  Escherichia coli

<400>  4

Met Arg Thr Leu Gln Gly Trp Leu Leu Pro Val Phe Met Leu Pro Met 
1               5                   10                  15      


Ala Val Tyr Ala Gln Glu Ala Thr Val Lys Glu Val His Asp Ala Pro 
            20                  25                  30          


Ala Val Arg Gly Ser Ile Ile Ala Asn Met Leu Gln Glu His Asp Asn 
        35                  40                  45              


Pro Phe Thr Leu Tyr Pro Tyr Asp Thr Asn Tyr Leu Ile Tyr Thr Gln 
    50                  55                  60                  


Thr Ser Asp Leu Asn Lys Glu Ala Ile Ala Ser Tyr Asp Trp Ala Glu 
65                  70                  75                  80  


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


Leu Trp Arg Gly Ile Leu Gly Pro Asn Ser Val Leu Gly Ala Ser Tyr 
            100                 105                 110         


Thr Gln Lys Ser Trp Trp Gln Leu Ser Asn Ser Glu Glu Ser Ser Pro 
        115                 120                 125             


Phe Arg Glu Thr Asn Tyr Glu Pro Gln Leu Phe Leu Gly Phe Ala Thr 
    130                 135                 140                 


Asp Tyr Arg Phe Ala Gly Trp Thr Leu Arg Asp Val Glu Met Gly Tyr 
145                 150                 155                 160 


Asn His Asp Ser Asn Gly Arg Ser Asp Pro Thr Ser Arg Ser Trp Asn 
                165                 170                 175     


Arg Leu Tyr Thr Arg Leu Met Ala Glu Asn Gly Asn Trp Leu Val Glu 
            180                 185                 190         


Val Lys Pro Trp Tyr Val Val Gly Asn Thr Asp Asp Asn Pro Asp Ile 
        195                 200                 205             


Thr Lys Tyr Met Gly Tyr Tyr Gln Leu Lys Ile Gly Tyr His Leu Gly 
    210                 215                 220                 


Asp Ala Val Leu Ser Ala Lys Gly Gln Tyr Asn Trp Asn Thr Gly Tyr 
225                 230                 235                 240 


Gly Gly Ala Glu Leu Gly Leu Ser Tyr Pro Ile Thr Lys His Val Arg 
                245                 250                 255     


Leu Tyr Thr Gln Val Tyr Ser Gly Tyr Gly Glu Ser Leu Ile Asp Tyr 
            260                 265                 270         


Asn Phe Asn Gln Thr Arg Val Gly Val Gly Val Met Leu Asn Asp Leu 
        275                 280                 285             


Phe 
    


<210>  5
<211>  208
<212>  PRT
<213>  Escherichia coli

<400>  5

Met Met Asn Phe Asn Asn Val Phe Arg Trp His Leu Pro Phe Leu Phe 
1               5                   10                  15      


Leu Val Leu Leu Thr Phe Arg Ala Ala Ala Ala Asp Thr Leu Leu Ile 
            20                  25                  30          


Leu Gly Asp Ser Leu Ser Ala Gly Tyr Arg Met Ser Ala Ser Ala Ala 
        35                  40                  45              


Trp Pro Ala Leu Leu Asn Asp Lys Trp Gln Ser Lys Thr Ser Val Val 
    50                  55                  60                  


Asn Ala Ser Ile Ser Gly Asp Thr Ser Gln Gln Gly Leu Ala Arg Leu 
65                  70                  75                  80  


Pro Ala Leu Leu Lys Gln His Gln Pro Arg Trp Val Leu Val Glu Leu 
                85                  90                  95      


Gly Gly Asn Asp Gly Leu Arg Gly Phe Gln Pro Gln Gln Thr Glu Gln 
            100                 105                 110         


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


Leu Leu Met Gln Ile Arg Leu Pro Ala Asn Tyr Gly Arg Arg Tyr Asn 
    130                 135                 140                 


Glu Ala Phe Ser Ala Ile Tyr Pro Lys Leu Ala Lys Glu Phe Asp Val 
145                 150                 155                 160 


Pro Leu Leu Pro Phe Phe Met Glu Glu Val Tyr Leu Lys Pro Gln Trp 
                165                 170                 175     


Met Gln Asp Asp Gly Ile His Pro Asn Arg Asp Ala Gln Pro Phe Ile 
            180                 185                 190         


Ala Asp Trp Met Ala Lys Gln Leu Gln Pro Leu Val Asn His Asp Ser 
        195                 200                 205             


<210>  6
<211>  345
<212>  PRT
<213>  Haemophilus influenzae

<400>  6

Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys Ser Asp Lys 
1               5                   10                  15      


Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro Glu His Thr 
            20                  25                  30          


Leu Glu Ser Lys Ala Leu Ala Phe Ala Gln Gln Ala Asp Tyr Leu Glu 
        35                  40                  45              


Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val Ile His Asp 
    50                  55                  60                  


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


His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr Leu Lys Glu 
                85                  90                  95      


Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Lys Asp Gly Lys 
            100                 105                 110         


Gln Ala Gln Val Tyr Pro Asn Arg Phe Pro Leu Trp Lys Ser His Phe 
        115                 120                 125             


Arg Ile His Thr Phe Glu Asp Glu Ile Glu Phe Ile Gln Gly Leu Glu 
    130                 135                 140                 


Lys Ser Thr Gly Lys Lys Val Gly Ile Tyr Pro Glu Ile Lys Ala Pro 
145                 150                 155                 160 


Trp Phe His His Gln Asn Gly Lys Asp Ile Ala Ala Glu Thr Leu Lys 
                165                 170                 175     


Val Leu Lys Lys Tyr Gly Tyr Asp Lys Lys Thr Asp Met Val Tyr Leu 
            180                 185                 190         


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


Pro Gln Met Gly Met Asp Leu Lys Leu Val Gln Leu Ile Ala Tyr Thr 
    210                 215                 220                 


Asp Trp Lys Glu Thr Gln Glu Lys Asp Pro Lys Gly Tyr Trp Val Asn 
225                 230                 235                 240 


Tyr Asn Tyr Asp Trp Met Phe Lys Pro Gly Ala Met Ala Glu Val Val 
                245                 250                 255     


Lys Tyr Ala Asp Gly Val Gly Pro Gly Trp Tyr Met Leu Val Asn Lys 
            260                 265                 270         


Glu Glu Ser Lys Pro Asp Asn Ile Val Tyr Thr Pro Leu Val Lys Glu 
        275                 280                 285             


Leu Ala Gln Tyr Asn Val Glu Val His Pro Tyr Thr Val Arg Lys Asp 
    290                 295                 300                 


Ala Leu Pro Glu Phe Phe Thr Asp Val Asn Gln Met Tyr Asp Ala Leu 
305                 310                 315                 320 


Leu Asn Lys Ser Gly Ala Thr Gly Val Phe Thr Asp Phe Pro Asp Thr 
                325                 330                 335     


Gly Val Glu Phe Leu Lys Gly Ile Lys 
            340                 345 


<210>  7
<211>  95
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CRM197 Expression Plasmid Sequence

<400>  7
aaactgagcc tgttctttga gatcaaaagc taatcggctg ctaacaaagc ccgaaaggaa       60

gctgagttgg ctgctgccac cgctgctggt tcgct                                  95


<210>  8
<211>  109
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CRM197 Expression Plasmid Derivative 1

<400>  8
aaactgagcc tgttctttga gatcaaaagc taatcgatat cgcaagcttt aagaggacaa       60

cattatgcgg actctgcagg gctggttgtt gccggtgttt atgttgcct                  109


<210>  9
<211>  108
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CRM197 Expression Plasmid Derivative 2

<400>  9
aaactgagcc tgttctttga gatcaaaagc taatcgatat cgcaatcttt aaaggacaac       60

attatgcgga ctctgcaggg ctggttgttg ccggtgttta tgttgcct                   108


