                         SEQUENCE LISTING

<110>  PETIVA PRIVATE LIMITED
 
<120>  A NOVEL WHOLE CELL BIOCATALYST FOR THE PRODUCTION OF TREHALULOSE

<130>  PCT1508

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  2070
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Gene sequence encoding for cell surface anchor protein fused to 
       C-terminus of  sucrose isomerase

<400>  1
atgcagttac ttcgctgttt ttcaatattt tctgttattg cttcagtttt agcacaggaa       60

ctgacaacta tatgcgagca aatcccctca ccaactttag aatcgacgcc gtactctttg      120

tcaacgacta ctattttggc caacgggaag gcaatgcaag gagtttttga atattacaaa      180

tcagtaacgt ttgtcagtaa ttgcggttct cacccctcaa caactagcaa aggcagcccc      240

ataaacacac agtatgtttt taaggacaat agctcgacga ttgaaggtag atacccatac      300

gacgttccag actacgctct gcaggctagt ggtggaggag gctctggtgg aggcggtagc      360

ggaggcggag ggtcggctag catggaagaa gccgtcaaac cgggtgctcc gtggtggaaa      420

tccgcagtgt tttatcaagt ctatccgcgt tcattcaaag atacgaatgg tgatggcatt      480

ggtgatttta aaggtctgac cgaaaaactg gattatctga aaggcctggg tatcgatgcc      540

atttggatca acccgcatta cgcatctccg aacacggata atggctatga tattagtgat      600

taccgcgaag ttatgaaaga atatggtacc atggaagatt tcgatcgtct gatggcggaa      660

ctgaaaaaac gtggcatgcg cctgatggtg gatgtggtta tcaaccatag ctctgatcag      720

cacgaatggt ttaaaagtag ccgcgccagc aaagataatc cgtatcgtga ttattacttc      780

tggcgcgatg gcaaagatgg tcatgaaccg aacaattacc cgagcttttt cggcggtagc      840

gcgtgggaaa aagatccggt gacgggccag tattacctgc actattttgg tcgccagcag      900

ccggatctga actgggatac cccgaaactg cgcgaagaac tgtacgccat gctgcgtttt      960

tggctggata aaggcgttag tggtatgcgt ttcgatacgg tggcaaccta tagcaaaacg     1020

ccgggctttc cggatctgac cccggaacag atgaaaaact tcgccgaagc atatacccag     1080

ggtccgaatc tgcatcgtta cctgcaggaa atgcatgaaa aagttttcga tcactacgat     1140

gccgtgaccg caggcgaaat ttttggtgcg ccgctgaatc aggttccgct gttcatcgat     1200

agccgtcgca aagaactgga tatggcgttt accttcgatc tgattcgtta tgatcgcgcc     1260

ctggatcgtt ggcacacgat cccgcgcacc ctggcagatt ttcgtcagac gattgataaa     1320

gtggatgcga tcgccggcga atacggttgg aacacctttt tcctgggcaa ccatgataat     1380

ccgcgcgccg tttctcactt cggtgatgat cgtccgcagt ggcgcgaagc aagtgcgaaa     1440

gccctggcaa ccgtgacgct gacccagcgt ggtaccccgt ttattttcca gggcgatgaa     1500

ctgggtatga cgaattatcc gtttaaaacc ctgcaggatt tcgatgatat cgaagttaaa     1560

ggctttttcc aggattacgt ggaaacgggt aaagcgaccg ccgaagaact gctgacgaac     1620

gttgcactga ccagccgtga taatgcgcgc accccgtttc agtgggatga ttctgcaaat     1680

gcgggcttca ccacgggtaa accgtggctg aaagttaacc cgaattatac cgaaattaac     1740

gcggcccgcg aaatcggcga tccgaaatct gtgtatagtt tttaccgcaa tctgattagc     1800

atccgtcatg aaacgccggc actgagcacc ggttcttatc gtgatattga tccgagcaac     1860

gcggatgtgt atgcctacac gcgttctcag gatggtgaaa cctacctggt ggttgtgaat     1920

tttaaagcag aaccgcgtag cttcacgctg ccggatggca tgcacattgc ggaaaccctg     1980

atcgaatcta gtagcccggc agcgccggcg gctggtgcgg cttcgctgga actgcaaccg     2040

tggcaaagtg gtatttacaa agtgaaataa                                      2070


<210>  2
<211>  772
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GAPDH promoter sequence

<400>  2
tcaacaacaa gaagtttaat gacgcggagg ccaaggcaaa aagattcctt gattacgtaa       60

gggagttaga atcattttga ataaaaaaca cgctttttca gttcgagttt atcattatca      120

atactgccat ttcaaagaat acgtaaataa ttaatagtag tgattttcct aactttattt      180

agtcaaaaaa ttagcctttt aattctgctg taacccgtac atgcccaaaa tagggggcgg      240

gttacacaga atatataaca tcgtaggtgt ctgggtgaac agtttattcc tggcatccac      300

taaatataat ggagcccgct ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc      360

aaaatattgt tttcttcacc aaccatcagt tcataggtcc attctcttag cgcaactaca      420

gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct caatggagtg atgcaacctg      480

cctggagtaa atgatgacac aaggcaattg acccacgcat gtatctatct cattttctta      540

caccttctat taccttctgc tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac      600

cagttccctg aaattattcc cctacttgac taataagtat ataaagacgg taggtattga      660

ttgtaattct gtaaatctat ttcttaaact tcttaaattc tacttttata gttagtcttt      720

tttttagttt taaaacacca agaacttagt ttcgaataaa cacacataaa ca              772


<210>  3
<211>  451
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GAL1 promoter sequence

<400>  3
acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt       60

cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga      120

acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac      180

ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga      240

ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat      300

taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc      360

ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac      420

ctctatactt taacgtcaag gagaaaaaac c                                     451


<210>  4
<211>  125
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GAPDH terminater sequence

<400>  4
ttggttgaac acgttgccaa ggcttaagtg aatttacttt aaatcttgca tttaaataaa       60

ttttcttttt atagctttat gacttagttt caatttatat actattttaa tgacattttc      120

gattc                                                                  125


<210>  5
<211>  305
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Alpha Factor terminater sequence

<400>  5
ctgataacaa cagtgtagat gtaacaaaat cgactttgtt cccactgtac ttttagctcg       60

tacaaaatac aatatacttt tcatttctcc gtaaacaaca tgttttccca tgtaatatcc      120

ttttctattt ttcgttccgt taccaacttt acacatactt tatatagcta ttcacttcta      180

tacactaaaa aactaagaca attttaattt tgctgcctgc catatttcaa tttgttataa      240

attcctataa tttatcctat tagtagctaa aaaaagatga atgtgaatcg aatcctaaga      300

gaatt                                                                  305


<210>  6
<211>  689
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of cell surface anchor and sucrose isomerase 
       fusion protein

<400>  6

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val 
1               5                   10                  15      


Leu Ala Gln Glu Leu Thr Thr Ile Cys Glu Gln Ile Pro Ser Pro Thr 
            20                  25                  30          


Leu Glu Ser Thr Pro Tyr Ser Leu Ser Thr Thr Thr Ile Leu Ala Asn 
        35                  40                  45              


Gly Lys Ala Met Gln Gly Val Phe Glu Tyr Tyr Lys Ser Val Thr Phe 
    50                  55                  60                  


Val Ser Asn Cys Gly Ser His Pro Ser Thr Thr Ser Lys Gly Ser Pro 
65                  70                  75                  80  


Ile Asn Thr Gln Tyr Val Phe Lys Asp Asn Ser Ser Thr Ile Glu Gly 
                85                  90                  95      


Arg Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Leu Gln Ala Ser Gly Gly 
            100                 105                 110         


Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser Met 
        115                 120                 125             


Glu Glu Ala Val Lys Pro Gly Ala Pro Trp Trp Lys Ser Ala Val Phe 
    130                 135                 140                 


Tyr Gln Val Tyr Pro Arg Ser Phe Lys Asp Thr Asn Gly Asp Gly Ile 
145                 150                 155                 160 


Gly Asp Phe Lys Gly Leu Thr Glu Lys Leu Asp Tyr Leu Lys Gly Leu 
                165                 170                 175     


Gly Ile Asp Ala Ile Trp Ile Asn Pro His Tyr Ala Ser Pro Asn Thr 
            180                 185                 190         


Asp Asn Gly Tyr Asp Ile Ser Asp Tyr Arg Glu Val Met Lys Glu Tyr 
        195                 200                 205             


Gly Thr Met Glu Asp Phe Asp Arg Leu Met Ala Glu Leu Lys Lys Arg 
    210                 215                 220                 


Gly Met Arg Leu Met Val Asp Val Val Ile Asn His Ser Ser Asp Gln 
225                 230                 235                 240 


His Glu Trp Phe Lys Ser Ser Arg Ala Ser Lys Asp Asn Pro Tyr Arg 
                245                 250                 255     


Asp Tyr Tyr Phe Trp Arg Asp Gly Lys Asp Gly His Glu Pro Asn Asn 
            260                 265                 270         


Tyr Pro Ser Phe Phe Gly Gly Ser Ala Trp Glu Lys Asp Pro Val Thr 
        275                 280                 285             


Gly Gln Tyr Tyr Leu His Tyr Phe Gly Arg Gln Gln Pro Asp Leu Asn 
    290                 295                 300                 


Trp Asp Thr Pro Lys Leu Arg Glu Glu Leu Tyr Ala Met Leu Arg Phe 
305                 310                 315                 320 


Trp Leu Asp Lys Gly Val Ser Gly Met Arg Phe Asp Thr Val Ala Thr 
                325                 330                 335     


Tyr Ser Lys Thr Pro Gly Phe Pro Asp Leu Thr Pro Glu Gln Met Lys 
            340                 345                 350         


Asn Phe Ala Glu Ala Tyr Thr Gln Gly Pro Asn Leu His Arg Tyr Leu 
        355                 360                 365             


Gln Glu Met His Glu Lys Val Phe Asp His Tyr Asp Ala Val Thr Ala 
    370                 375                 380                 


Gly Glu Ile Phe Gly Ala Pro Leu Asn Gln Val Pro Leu Phe Ile Asp 
385                 390                 395                 400 


Ser Arg Arg Lys Glu Leu Asp Met Ala Phe Thr Phe Asp Leu Ile Arg 
                405                 410                 415     


Tyr Asp Arg Ala Leu Asp Arg Trp His Thr Ile Pro Arg Thr Leu Ala 
            420                 425                 430         


Asp Phe Arg Gln Thr Ile Asp Lys Val Asp Ala Ile Ala Gly Glu Tyr 
        435                 440                 445             


Gly Trp Asn Thr Phe Phe Leu Gly Asn His Asp Asn Pro Arg Ala Val 
    450                 455                 460                 


Ser His Phe Gly Asp Asp Arg Pro Gln Trp Arg Glu Ala Ser Ala Lys 
465                 470                 475                 480 


Ala Leu Ala Thr Val Thr Leu Thr Gln Arg Gly Thr Pro Phe Ile Phe 
                485                 490                 495     


Gln Gly Asp Glu Leu Gly Met Thr Asn Tyr Pro Phe Lys Thr Leu Gln 
            500                 505                 510         


Asp Phe Asp Asp Ile Glu Val Lys Gly Phe Phe Gln Asp Tyr Val Glu 
        515                 520                 525             


Thr Gly Lys Ala Thr Ala Glu Glu Leu Leu Thr Asn Val Ala Leu Thr 
    530                 535                 540                 


Ser Arg Asp Asn Ala Arg Thr Pro Phe Gln Trp Asp Asp Ser Ala Asn 
545                 550                 555                 560 


Ala Gly Phe Thr Thr Gly Lys Pro Trp Leu Lys Val Asn Pro Asn Tyr 
                565                 570                 575     


Thr Glu Ile Asn Ala Ala Arg Glu Ile Gly Asp Pro Lys Ser Val Tyr 
            580                 585                 590         


Ser Phe Tyr Arg Asn Leu Ile Ser Ile Arg His Glu Thr Pro Ala Leu 
        595                 600                 605             


Ser Thr Gly Ser Tyr Arg Asp Ile Asp Pro Ser Asn Ala Asp Val Tyr 
    610                 615                 620                 


Ala Tyr Thr Arg Ser Gln Asp Gly Glu Thr Tyr Leu Val Val Val Asn 
625                 630                 635                 640 


Phe Lys Ala Glu Pro Arg Ser Phe Thr Leu Pro Asp Gly Met His Ile 
                645                 650                 655     


Ala Glu Thr Leu Ile Glu Ser Ser Ser Pro Ala Ala Pro Ala Ala Gly 
            660                 665                 670         


Ala Ala Ser Leu Glu Leu Gln Pro Trp Gln Ser Gly Ile Tyr Lys Val 
        675                 680                 685             


Lys 
    


<210>  7
<211>  1357
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Deletion cassette used for disruption of SUC2 gene

<400>  7
gttcttgtgc ttttttttta ccatatatct tacttttttt tttctctcag agaaacaagc       60

aaaacaaaaa gcttttcttt tcactaacgt acagctgaag cttcgtacgc tgcaggtcga      120

caacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt ctagagatct      180

gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc      240

ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt      300

tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg      360

cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca      420

gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg      480

ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac      540

atcacatccg aacataaaca accatggccg accaagcgac gcccaacctg ccatcacgag      600

atttcgattc cacggccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg      660

ccggctggat gatcctccag cgcggggatc tcaagctgga gttcttcgcc caccccgggc      720

tcgatcccct cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct      780

accggcagtg caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg      840

cccccgaact gcaggagtgg ggaggcacga tggccgcttt ggtcgacccg gacgggacgc      900

tcctgcgcct gatacagaac gaattgcttg caggcatctc atgatcagta ctgacaataa      960

aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct     1020

attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga     1080

tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc     1140

tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctctcgaga     1200

acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtga tatcagatcc     1260

actagtggcc tatgcattct aaagggcttt agctaacgag tgacgaatgt aaaactttat     1320

gatttcaaag aatacctcca aaccattgaa aatgtat                              1357


<210>  8
<211>  1793
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Deletion cassette used for disruption of AGT1gene

<400>  8
ggtattgatt gtttgaagaa ttttcgggtt ggtgtttctt tctgatgcta catagaagaa       60

catcaaacaa ctaaaaaaat agtataatac agctgaagct tcgtacgctg caggtcgaca      120

acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtct agagatctgt      180

ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct      240

ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta      300

gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg      360

aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga      420

cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc      480

actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat      540

cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag gccgcgatta      600

aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa      660

tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa      720

catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg      780

acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg      840

ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat      900

tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct      960

gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga     1020

atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt     1080

gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact     1140

catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt     1200

gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc     1260

ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat     1320

cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagtactg     1380

acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag     1440

ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct     1500

gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc     1560

tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacgagct     1620

ctcgagaacc cttaatataa cttcgtataa tgtatgctat acgaagttat taggtgatat     1680

cagatccact agtggcctat gcagtagtta gttaaaatag cccaagcagc aatcaagcaa     1740

atatgagagt actttttctt tagcacctgg tacttgtgcc tggatattga ttc            1793


