               SEQUENCE LISTING

<110> Glycom A/S

<120> NEW MAJOR FACILITATOR SUPERFAMILY (MFS) PROTEIN (BAD) IN HMO PRODUCTION

<130> P2914WO00

<150> DK202000087
<151> 2020-01-23

<160> 31

<170> BiSSAP 1.3.6

<210> 1
<211> 387
<212> PRT
<213> Rouxiella badensis


<220> 
<223> bad_WP_017489914.1\optimized translation:

<400> 1
Met Ser Ser Arg Arg Leu Ser Ile Ile Phe Ala Thr Phe Leu Leu Val 
1               5                   10                  15      
Ser Phe Leu Thr Gly Ile Ala Gly Ala Leu Gln Ala Pro Thr Leu Ser 
            20                  25                  30          
Leu Phe Leu Thr Asn Glu Val Lys Val Arg Pro Leu Trp Val Gly Leu 
        35                  40                  45              
Phe Tyr Thr Val Asn Ala Leu Gly Gly Ile Val Ile Ser Phe Leu Leu 
    50                  55                  60                  
Ala Asn Tyr Ser Asp Lys Lys Gly Asp Arg Arg Lys Leu Leu Phe Phe 
65                  70                  75                  80  
Cys Thr Leu Met Ala Ile Gly Asn Ser Leu Ile Phe Ala Tyr Ser Arg 
                85                  90                  95      
Asp Tyr Leu Val Leu Ile Ser Val Gly Val Leu Leu Ala Ala Ile Gly 
            100                 105                 110         
Asn Ala Ser Met Pro Gln Leu Phe Ala Leu Ala Arg Glu Tyr Ala Asp 
        115                 120                 125             
Arg Ser Ala His Glu Val Val Met Phe Ser Ser Met Met Arg Ala Thr 
    130                 135                 140                 
Leu Ser Leu Ala Trp Val Leu Gly Pro Pro Ile Ser Phe Thr Leu Ala 
145                 150                 155                 160 
Leu Asn Tyr Gly Phe Thr Leu Met Tyr Leu Cys Ala Ala Gly Val Phe 
                165                 170                 175     
Ile Phe Ser Ala Leu Met Val Trp Phe Phe Leu Pro Ser Val Gly Arg 
            180                 185                 190         
Ile Glu Gln Pro Val Asp Lys Val Val Val His Val Ser Ala Trp Lys 
        195                 200                 205             
Asn Arg Asp Val Arg Leu Leu Phe Phe Ala Ser Leu Leu Met Trp Thr 
    210                 215                 220                 
Cys Asn Ile Met Tyr Ile Ile Asp Met Pro Leu Tyr Ile Thr Ser Asp 
225                 230                 235                 240 
Leu Gly Leu Pro Glu Gly Leu Ala Gly Leu Leu Met Gly Ala Ala Ala 
                245                 250                 255     
Gly Leu Glu Ile Pro Val Met Leu Ile Ala Gly Tyr Leu Val Lys Arg 
            260                 265                 270         
Thr Gly Lys Arg Arg Leu Met Leu Cys Ala Ala Val Phe Gly Ile Leu 
        275                 280                 285             
Phe Tyr Leu Gly Leu Val Leu Phe Gln Phe Lys Ala Ala Leu Met Ile 
    290                 295                 300                 
Leu Gln Leu Phe Asn Ala Ile Phe Ile Gly Ile Ile Ala Gly Ile Gly 
305                 310                 315                 320 
Met Leu Tyr Phe Gln Asp Leu Met Pro Gly Arg Ala Gly Ser Ala Thr 
                325                 330                 335     
Thr Leu Phe Thr Asn Ser Ile Ser Thr Gly Ala Ile Leu Ala Gly Val 
            340                 345                 350         
Ile Gln Gly Thr Ile Val Gln Asn Phe Gly His Tyr Gln Val Tyr Trp 
        355                 360                 365             
Met Ala Leu Ala Leu Ala Val Gly Ala Leu Val Leu Met Thr Arg Val 
    370                 375                 380                 
Lys Asn Val 
385         

<210> 2
<211> 1164
<212> DNA
<213> Rouxiella badensis


<220> 
<223> bad nucleotide sequence

<400> 2
atgagcagcc gtcgtctgag catcattttc gcgacctttc tgctggttag cttcctgacc      60

ggtattgcgg gtgcgctgca ggcgccgacc ctgagcctgt ttctgaccaa cgaggttaaa     120

gtgcgtccgc tgtgggttgg tctgttctac accgtgaacg cgctgggtgg catcgttatt     180

agctttctgc tggcgaacta tagcgacaag aaaggtgatc gtcgtaaact gctgttcttt     240

tgcaccctga tggcgatcgg caacagcctg attttcgcgt acagccgtga ctatctggtg     300

ctgatcagcg ttggtgtgct gctggcggcg attggcaacg cgagcatgcc gcagctgttt     360

gcgctggcgc gtgagtacgc ggatcgtagc gcgcacgaag tggttatgtt tagcagcatg     420

atgcgtgcga ccctgagcct ggcgtgggtt ctgggtccgc cgatcagctt caccctggcg     480

ctgaactacg gttttaccct gatgtatctg tgcgcggcgg gcgtgttcat ctttagcgcg     540

ctgatggttt ggttctttct gccgagcgtg ggccgtattg aacaaccggt tgacaaggtg     600

gttgtgcacg tgagcgcgtg gaaaaaccgt gatgttcgtc tgctgttctt tgcgagcctg     660

ctgatgtgga cctgcaacat catgtacatc attgacatgc cgctgtatat taccagcgat     720

ctgggtctgc cggagggtct ggcgggcctg ctgatgggtg ctgcggcggg cctggaaatc     780

ccggttatgc tgattgcggg ttacctggtg aagcgtaccg gcaaacgtcg tctgatgctg     840

tgcgcggcgg ttttcggtat cctgttttat ctgggcctgg tgctgttcca gtttaaggcg     900

gcgctgatga tcctgcaact gttcaacgcg atctttattg gtatcattgc gggtattggc     960

atgctgtatt tccaggacct gatgccgggt cgtgcgggca gcgcgaccac cctgtttacc    1020

aacagcatca gcaccggcgc gattctggcg ggtgttatcc agggcaccat tgtgcaaaac    1080

ttcggtcact accaagtgta ttggatggcg ctggcgctgg cggttggtgc gctggtgctg    1140

atgacccgtg ttaaaaacgt gtaa                                           1164


<210> 3
<211> 291
<212> DNA
<213> Artificial Sequence


<220> 
<223> Promoter sequence PgatY_70UTR

<400> 3
cggcaaccta tgcctgatgc gacgctgaag cgtcttatca tgcctacata gcactgccac     60

gtatgtttac accgcatccg gcataaaaac acgcgcactt tgctacggct tccctatcgg    120

gaggccgttt ttttgccttt cactcctcga ataattttca tattgtcgtt tttgtgatcg    180

ttatctcgat atttaaaaac aaataatttc attatatttt gtgcctacaa gcatcgtgga    240

ggtccgtgac tttcacgcat acaacaaaca ttaaccaagg aggaaacagc t             291


<210> 4
<211> 300
<212> DNA
<213> Artificial Sequence


<220> 
<223> PglpF regulatory sequence

<300> 
<310> WO2019123324
<311> 19.12.2018
<312> 27.06.2019
<313> 1-300

<400> 4
gcggcacgcc ttgcagatta cggtttgcca cacttttcat ccttctcctg gtgacataat     60

ccacatcaat cgaaaatgtt aataaatttg ttgcgcgaat gatctaacaa acatgcatca    120

tgtacaatca gatggaataa atggcgcgat aacgctcatt ttatgacgag gcacacacat    180

tttaagttcg atatttctcg tttttgctcg ttaacgataa gtttacagca tgcctacaag    240

catcgtggag gtccgtgact ttcacgcata caacaaacat taaccaagga ggaaacagct    300


<210> 5
<211> 300
<212> DNA
<213> Artificial Sequence


<220> 
<223> PglpF_SD4 regulatory sequence

<300> 
<310> WO2019123324
<311> 19.12.2018
<312> 27.06.2019
<313> 1-300

<400> 5
gcggcacgcc ttgcagatta cggtttgcca cacttttcat ccttctcctg gtgacataat     60

ccacatcaat cgaaaatgtt aataaatttg ttgcgcgaat gatctaacaa acatgcatca    120

tgtacaatca gatggaataa atggcgcgat aacgctcatt ttatgacgag gcacacacat    180

tttaagttcg atatttctcg tttttgctcg ttaacgataa gtttacagca tgcctacaag    240

catcgtggag gtccgtgact ttcacgcata caacaaacat taaccaacta ggaaacagct    300


<210> 6
<211> 203
<212> DNA
<213> Artificial Sequence


<220> 
<223> Promoter sequence PmglB_70UTR

<400> 6
tgcgtcgcca ttctgtcgca acacgccaga atgcggcggc gatcactaac tcaacaaatc     60

aggcgatgta accgctttca atctgtgagt gatttcacag tatcttaaca atgtgatagc    120

tatgattgca ccgtgcctac aagcatcgtg gaggtccgtg actttcacgc atacaacaaa    180

cattaaccaa ggaggaaaca gct                                            203


<210> 7
<211> 203
<212> DNA
<213> Artificial Sequence


<220> 
<223> Promoter sequence PmglB_70UTR_SD4

<400> 7
tgcgtcgcca ttctgtcgca acacgccaga atgcggcggc gatcactaac tcaacaaatc     60

aggcgatgta accgctttca atctgtgagt gatttcacag tatcttaaca atgtgatagc    120

tatgattgca ccgtgcctac aagcatcgtg gaggtccgtg actttcacgc atacaacaaa    180

cattaaccaa ctaggaaaca gct                                            203


<210> 8
<211> 34
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O40 backbone.for

<220> 
<221> misc_RNA
<222> 9

<400> 8
attaacccuc caggcatcaa ataaaacgaa aggc                                 34


<210> 9
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O79 Backbone.rev

<220> 
<221> misc_RNA
<222> 10

<400> 9
atttgcgcau caccaatcaa attcacgcgg cc                                   32


<210> 10
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O261, PglpF.for

<220> 
<221> misc_RNA
<222> 10

<400> 10
atgcgcaaau gcggcacgcc ttgcagatta cg                                   32


<210> 11
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O262, PglpF.rev

<220> 
<221> misc_RNA
<222> 8

<400> 11
agctgttucc tccttggtta atgtttgttg tatgcg                               36


<210> 12
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O459, PglpF_SD4

<220> 
<221> misc_RNA
<222> 8

<400> 12
agctgttucc tagttggtta atgtttgttg tatgcg                               36


<210> 13
<211> 29
<212> DNA
<213> Artificial Sequence


<220> 
<223> primer KABY729, bad.for

<220> 
<221> misc_RNA
<222> 8

<400> 13
aaacagcuat gagcagccgt cgtctgagc                                      29


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


<220> 
<223> Primer KABY730, bad.rev

<220> 
<221> misc_RNA
<222> 9

<400> 14
agggttaaut tacacgtttt taacacgggt catcag                               36


<210> 15
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O48, galK.for

<400> 15
cccagcgaga cctgaccgca gaac                                           24


<210> 16
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O49, glaK.rev

<400> 16
ccccagtcca tcagcgtgac tacc                                           24


<210> 17
<211> 195
<212> DNA
<213> Artificial Sequence


<220> 
<223> Plac expression element

<300> 
<310> WO2019123324
<311> 19.12.2018
<312> 27.06.2019

<400> 17
atgcgcaaat tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc     60

cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg    120

accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctgggtacct    180

aaggaggaaa cagct                                                     195


<210> 18
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O68, Plac.for

<220> 
<221> misc_RNA
<222> 10

<400> 18
atgcgcaaau tgtgagttag ctcactcatt ag                                   32


<210> 19
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer O133, Plac.rev

<220> 
<221> misc_RNA
<222> 8

<400> 19
agctgttucc tccttaggta cccagctttt gttccc                               36


<210> 20
<211> 33
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY745, vag.for

<400> 20
aaacagcuat gaagagcctg ctgacccgta aac                                  33


<210> 21
<211> 31
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY746, vag.rev

<220> 
<221> misc_RNA
<222> 9

<400> 21
agggttaaut taaacgtttt tcacacgcgc g                                   31


<210> 22
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY721, yberC0001_9420.for

<220> 
<221> misc_RNA
<222> 8

<400> 22
aaacagcuat gaagagcgcg ctgaccttta gc                                   32


<210> 23
<211> 29
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY722, yberC0001_9420.rev

<220> 
<221> misc_RNA
<222> 9

<400> 23
agggttaaut tacgcctcac gcacacgcg                                      29


<210> 24
<211> 31
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY733, fred.for

<220> 
<221> misc_RNA
<222> 8

<400> 24
aaacagcuat gaagagcgcg ctgaccttca g                                   31


<210> 25
<211> 29
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY734, fred.rev

<220> 
<221> misc_RNA
<222> 9

<400> 25
agggttaaut tacgcttcac gcacacgcg                                      29


<210> 26
<211> 28
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY741, nec.for

<220> 
<221> misc_RNA
<222> 8

<400> 26
aaacagcuat gcagagcttc accccgcc                                       28


<210> 27
<211> 34
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY742, nec.rev

<220> 
<221> misc_RNA
<222> 9

<400> 27
agggttaaut tacgcctgct ctttaacacg cagc                                 34


<210> 28
<211> 31
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY737, marc.for

<220> 
<221> misc_RNA
<222> 8

<400> 28
aaacagcuat gcagcgtctg agccgtctga g                                   31


<210> 29
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer KABY738, marc.rev

<220> 
<221> misc_RNA
<222> 9

<400> 29
agggttaaut taaacttcac gcactttcgc gc                                   32


<210> 30
<211> 31
<212> DNA
<213> Artificial Sequence


<220> 
<223> Primer MP1217, yabM.for

<220> 
<221> misc_RNA
<222> 8

<400> 30
aaacagcuat gaaggcgctg tggagccgtc g                                   31


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


<220> 
<223> Primer MP1218, yabM.rev

<220> 
<221> misc_RNA
<222> 9

<400> 31
agggttaauc gccagcggaa cgctcttcac g                                   31


