                         SEQUENCE LISTING

<110>  CARIBOU BIOSCIENCES, INC.
 
<120>  ENGINEERED GUT MICROBES AND USES THEREOF

<130>  CBI028.30

<150>  US 62/626,586
<151>  2018-02-05

<160>  50    

<170>  PatentIn version 3.5

<210>  1
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 23 sgRNA

<400>  1
agcggataac aatttcacac                                                   20


<210>  2
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:1

<400>  2
gtgtgaaatt gttatccgct                                                   20


<210>  3
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 24 sgRNA

<400>  3
cgtcgccacc aatccccata                                                   20


<210>  4
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:3

<400>  4
tatggggatt ggtggcgacg                                                   20


<210>  5
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 64 sgRNA

<400>  5
cgttttacaa cgtcgtgact                                                   20


<210>  6
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:5

<400>  6
agtcacgacg ttgtaaaacg                                                   20


<210>  7
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 65 sgRNA

<400>  7
ggccagtgaa tccgtaatca                                                   20


<210>  8
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:7

<400>  8
tgattacgga ttcactggcc                                                   20


<210>  9
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 66 sgRNA

<400>  9
cttcttccgc gtgcagcaga                                                   20


<210>  10
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:9

<400>  10
tctgctgcac gcggaagaag                                                   20


<210>  11
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 67 sgRNA

<400>  11
ggcacatggc tgaatatcga                                                   20


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:11

<400>  12
tcgatattca gccatgtgcc                                                   20


<210>  13
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 13 sgRNA

<400>  13
cggcctgtgg gcattcagtc                                                   20


<210>  14
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:13

<400>  14
gactgaatgc ccacaggccg                                                   20


<210>  15
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 14 sgRNA

<400>  15
actgtggaat tgatcagcgt                                                   20


<210>  16
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:15

<400>  16
actgtggaat tgatcagcgt                                                   20


<210>  17
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 15 sgRNA

<400>  17
tatcgtgctg cgtttcgatg                                                   20


<210>  18
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:17

<400>  18
catcgaaacg cagcacgata                                                   20


<210>  19
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Oligo for generating guide 16 sgRNA

<400>  19
tttgaagccg atgtcacgcc                                                   20


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

<220>
<223>  Synthetic: Oligo complementary to SeqID NO:19

<400>  20
ggcgtgacat cggcttcaaa                                                   20


<210>  21
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 protein sequence

<400>  21

Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


<210>  22
<211>  3075
<212>  DNA
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(3075)
<223>  lacZ DNA sequence

<400>  22
atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct       60

ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc      120

gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc      180

tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct      240

gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc      300

tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg      360

acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg      420

cgaattattt ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc      480

ggttacggcc aggacagtcg tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc      540

ggagaaaacc gcctcgcggt gatggtgctg cgctggagtg acggcagtta tctggaagat      600

caggatatgt ggcggatgag cggcattttc cgtgacgtct cgttgctgca taaaccgact      660

acacaaatca gcgatttcca tgttgccact cgctttaatg atgatttcag ccgcgctgta      720

ctggaggctg aagttcagat gtgcggcgag ttgcgtgact acctacgggt aacagtttct      780

ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc      840

gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa      900

ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca caccgccgac      960

ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat     1020

ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg tcacgagcat     1080

catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatat cctgctgatg     1140

aagcagaaca actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac     1200

acgctgtgcg accgctacgg cctgtatgtg gtggatgaag ccaatattga aacccacggc     1260

atggtgccaa tgaatcgtct gaccgatgat ccgcgctggc taccggcgat gagcgaacgc     1320

gtaacgcgaa tggtgcagcg cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg     1380

aatgaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa atctgtcgat     1440

ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac cgatattatt     1500

tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc     1560

atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgatcctttg cgaatacgcc     1620

cacgcgatgg gtaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat     1680

ccccgtttac agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat     1740

gaaaacggca acccgtggtc ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc     1800

cagttctgta tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc gctgacggaa     1860

gcaaaacacc agcagcagtt tttccagttc cgtttatccg ggcaaaccat cgaagtgacc     1920

agcgaatacc tgttccgtca tagcgataac gagctcctgc actggatggt ggcgctggat     1980

ggtaagccgc tggcaagcgg tgaagtgcct ctggatgtcg ctccacaagg taaacagttg     2040

attgaactgc ctgaactacc gcagccggag agcgccgggc aactctggct cacagtacgc     2100

gtagtgcaac cgaacgcgac cgcatggtca gaagccgggc acatcagcgc ctggcagcag     2160

tggcgtctgg cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc catcccgcat     2220

ctgaccacca gcgaaatgga tttttgcatc gagctgggta ataagcgttg gcaatttaac     2280

cgccagtcag gctttctttc acagatgtgg attggcgata aaaaacaact gctgacgccg     2340

ctgcgcgatc agttcacccg tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc     2400

cgcattgacc ctaacgcctg ggtcgaacgc tggaaggcgg cgggccatta ccaggccgaa     2460

gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat tacgaccgct     2520

cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta ccggattgat     2580

ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac accgcatccg     2640

gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa ctggctcgga     2700

ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga ccgctgggat     2760

ctgccattgt cagacatgta taccccgtac gtcttcccga gcgaaaacgg tctgcgctgc     2820

gggacgcgcg aattgaatta tggcccacac cagtggcgcg gcgacttcca gttcaacatc     2880

agccgctaca gtcaacagca actgatggaa accagccatc gccatctgct gcacgcggaa     2940

gaaggcacat ggctgaatat cgacggtttc catatgggga ttggtggcga cgactcctgg     3000

agcccgtcag tatcggcgga attccagctg agcgccggtc gctaccatta ccagttggtc     3060

tggtgtcaaa aataa                                                      3075


<210>  23
<211>  1024
<212>  PRT
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(1024)
<223>  lacZ protein sequence

<400>  23

Met Thr Met Ile Thr Asp Ser Leu Ala Val Val Leu Gln Arg Arg Asp 
1               5                   10                  15      


Trp Glu Asn Pro Gly Val Thr Gln Leu Asn Arg Leu Ala Ala His Pro 
            20                  25                  30          


Pro Phe Ala Ser Trp Arg Asn Ser Glu Glu Ala Arg Thr Asp Arg Pro 
        35                  40                  45              


Ser Gln Gln Leu Arg Ser Leu Asn Gly Glu Trp Arg Phe Ala Trp Phe 
    50                  55                  60                  


Pro Ala Pro Glu Ala Val Pro Glu Ser Trp Leu Glu Cys Asp Leu Pro 
65                  70                  75                  80  


Glu Ala Asp Thr Val Val Val Pro Ser Asn Trp Gln Met His Gly Tyr 
                85                  90                  95      


Asp Ala Pro Ile Tyr Thr Asn Val Thr Tyr Pro Ile Thr Val Asn Pro 
            100                 105                 110         


Pro Phe Val Pro Thr Glu Asn Pro Thr Gly Cys Tyr Ser Leu Thr Phe 
        115                 120                 125             


Asn Val Asp Glu Ser Trp Leu Gln Glu Gly Gln Thr Arg Ile Ile Phe 
    130                 135                 140                 


Asp Gly Val Asn Ser Ala Phe His Leu Trp Cys Asn Gly Arg Trp Val 
145                 150                 155                 160 


Gly Tyr Gly Gln Asp Ser Arg Leu Pro Ser Glu Phe Asp Leu Ser Ala 
                165                 170                 175     


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


Ser Asp Gly Ser Tyr Leu Glu Asp Gln Asp Met Trp Arg Met Ser Gly 
        195                 200                 205             


Ile Phe Arg Asp Val Ser Leu Leu His Lys Pro Thr Thr Gln Ile Ser 
    210                 215                 220                 


Asp Phe His Val Ala Thr Arg Phe Asn Asp Asp Phe Ser Arg Ala Val 
225                 230                 235                 240 


Leu Glu Ala Glu Val Gln Met Cys Gly Glu Leu Arg Asp Tyr Leu Arg 
                245                 250                 255     


Val Thr Val Ser Leu Trp Gln Gly Glu Thr Gln Val Ala Ser Gly Thr 
            260                 265                 270         


Ala Pro Phe Gly Gly Glu Ile Ile Asp Glu Arg Gly Gly Tyr Ala Asp 
        275                 280                 285             


Arg Val Thr Leu Arg Leu Asn Val Glu Asn Pro Lys Leu Trp Ser Ala 
    290                 295                 300                 


Glu Ile Pro Asn Leu Tyr Arg Ala Val Val Glu Leu His Thr Ala Asp 
305                 310                 315                 320 


Gly Thr Leu Ile Glu Ala Glu Ala Cys Asp Val Gly Phe Arg Glu Val 
                325                 330                 335     


Arg Ile Glu Asn Gly Leu Leu Leu Leu Asn Gly Lys Pro Leu Leu Ile 
            340                 345                 350         


Arg Gly Val Asn Arg His Glu His His Pro Leu His Gly Gln Val Met 
        355                 360                 365             


Asp Glu Gln Thr Met Val Gln Asp Ile Leu Leu Met Lys Gln Asn Asn 
    370                 375                 380                 


Phe Asn Ala Val Arg Cys Ser His Tyr Pro Asn His Pro Leu Trp Tyr 
385                 390                 395                 400 


Thr Leu Cys Asp Arg Tyr Gly Leu Tyr Val Val Asp Glu Ala Asn Ile 
                405                 410                 415     


Glu Thr His Gly Met Val Pro Met Asn Arg Leu Thr Asp Asp Pro Arg 
            420                 425                 430         


Trp Leu Pro Ala Met Ser Glu Arg Val Thr Arg Met Val Gln Arg Asp 
        435                 440                 445             


Arg Asn His Pro Ser Val Ile Ile Trp Ser Leu Gly Asn Glu Ser Gly 
    450                 455                 460                 


His Gly Ala Asn His Asp Ala Leu Tyr Arg Trp Ile Lys Ser Val Asp 
465                 470                 475                 480 


Pro Ser Arg Pro Val Gln Tyr Glu Gly Gly Gly Ala Asp Thr Thr Ala 
                485                 490                 495     


Thr Asp Ile Ile Cys Pro Met Tyr Ala Arg Val Asp Glu Asp Gln Pro 
            500                 505                 510         


Phe Pro Ala Val Pro Lys Trp Ser Ile Lys Lys Trp Leu Ser Leu Pro 
        515                 520                 525             


Gly Glu Thr Arg Pro Leu Ile Leu Cys Glu Tyr Ala His Ala Met Gly 
    530                 535                 540                 


Asn Ser Leu Gly Gly Phe Ala Lys Tyr Trp Gln Ala Phe Arg Gln Tyr 
545                 550                 555                 560 


Pro Arg Leu Gln Gly Gly Phe Val Trp Asp Trp Val Asp Gln Ser Leu 
                565                 570                 575     


Ile Lys Tyr Asp Glu Asn Gly Asn Pro Trp Ser Ala Tyr Gly Gly Asp 
            580                 585                 590         


Phe Gly Asp Thr Pro Asn Asp Arg Gln Phe Cys Met Asn Gly Leu Val 
        595                 600                 605             


Phe Ala Asp Arg Thr Pro His Pro Ala Leu Thr Glu Ala Lys His Gln 
    610                 615                 620                 


Gln Gln Phe Phe Gln Phe Arg Leu Ser Gly Gln Thr Ile Glu Val Thr 
625                 630                 635                 640 


Ser Glu Tyr Leu Phe Arg His Ser Asp Asn Glu Leu Leu His Trp Met 
                645                 650                 655     


Val Ala Leu Asp Gly Lys Pro Leu Ala Ser Gly Glu Val Pro Leu Asp 
            660                 665                 670         


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


Pro Glu Ser Ala Gly Gln Leu Trp Leu Thr Val Arg Val Val Gln Pro 
    690                 695                 700                 


Asn Ala Thr Ala Trp Ser Glu Ala Gly His Ile Ser Ala Trp Gln Gln 
705                 710                 715                 720 


Trp Arg Leu Ala Glu Asn Leu Ser Val Thr Leu Pro Ala Ala Ser His 
                725                 730                 735     


Ala Ile Pro His Leu Thr Thr Ser Glu Met Asp Phe Cys Ile Glu Leu 
            740                 745                 750         


Gly Asn Lys Arg Trp Gln Phe Asn Arg Gln Ser Gly Phe Leu Ser Gln 
        755                 760                 765             


Met Trp Ile Gly Asp Lys Lys Gln Leu Leu Thr Pro Leu Arg Asp Gln 
    770                 775                 780                 


Phe Thr Arg Ala Pro Leu Asp Asn Asp Ile Gly Val Ser Glu Ala Thr 
785                 790                 795                 800 


Arg Ile Asp Pro Asn Ala Trp Val Glu Arg Trp Lys Ala Ala Gly His 
                805                 810                 815     


Tyr Gln Ala Glu Ala Ala Leu Leu Gln Cys Thr Ala Asp Thr Leu Ala 
            820                 825                 830         


Asp Ala Val Leu Ile Thr Thr Ala His Ala Trp Gln His Gln Gly Lys 
        835                 840                 845             


Thr Leu Phe Ile Ser Arg Lys Thr Tyr Arg Ile Asp Gly Ser Gly Gln 
    850                 855                 860                 


Met Ala Ile Thr Val Asp Val Glu Val Ala Ser Asp Thr Pro His Pro 
865                 870                 875                 880 


Ala Arg Ile Gly Leu Asn Cys Gln Leu Ala Gln Val Ala Glu Arg Val 
                885                 890                 895     


Asn Trp Leu Gly Leu Gly Pro Gln Glu Asn Tyr Pro Asp Arg Leu Thr 
            900                 905                 910         


Ala Ala Cys Phe Asp Arg Trp Asp Leu Pro Leu Ser Asp Met Tyr Thr 
        915                 920                 925             


Pro Tyr Val Phe Pro Ser Glu Asn Gly Leu Arg Cys Gly Thr Arg Glu 
    930                 935                 940                 


Leu Asn Tyr Gly Pro His Gln Trp Arg Gly Asp Phe Gln Phe Asn Ile 
945                 950                 955                 960 


Ser Arg Tyr Ser Gln Gln Gln Leu Met Glu Thr Ser His Arg His Leu 
                965                 970                 975     


Leu His Ala Glu Glu Gly Thr Trp Leu Asn Ile Asp Gly Phe His Met 
            980                 985                 990         


Gly Ile Gly Gly Asp Asp Ser Trp  Ser Pro Ser Val Ser  Ala Glu Phe 
        995                 1000                 1005             


Gln Leu  Ser Ala Gly Arg Tyr  His Tyr Gln Leu Val  Trp Cys Gln 
    1010                 1015                 1020             


Lys 
    


<210>  24
<211>  1812
<212>  DNA
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(1812)
<223>  gusA DNA sequence

<400>  24
atgttacgtc ctgtagaaac cccaacccgt gaaatcaaaa aactcgacgg cctgtgggca       60

ttcagtctgg atcgcgaaaa ctgtggaatt gatcagcgtt ggtgggaaag cgcgttacaa      120

gaaagccggg caattgctgt gccaggcagt tttaacgatc agttcgccga tgcagatatt      180

cgtaattatg cgggcaacgt ctggtatcag cgcgaagtct ttataccgaa aggttgggca      240

ggccagcgta tcgtgctgcg tttcgatgcg gtcactcatt acggcaaagt gtgggtcaat      300

aatcaggaag tgatggagca tcagggcggc tatacgccat ttgaagccga tgtcacgccg      360

tatgttattg ccgggaaaag tgtacgtatc accgtttgtg tgaacaacga actgaactgg      420

cagactatcc cgccgggaat ggtgattacc gacgaaaacg gcaagaaaaa gcagtcttac      480

ttccatgatt tctttaacta tgccgggatc catcgcagcg taatgctcta caccacgccg      540

aacacctggg tggacgatat caccgtggtg acgcatgtcg cgcaagactg taaccacgcg      600

tctgttgact ggcaggtggt ggccaatggt gatgtcagcg ttgaactgcg tgatgcggat      660

caacaggtgg ttgcaactgg acaaggcact agcgggactt tgcaagtggt gaatccgcac      720

ctctggcaac cgggtgaagg ttatctctat gaactgtgcg tcacagccaa aagccagaca      780

gagtgtgata tctacccgct tcgcgtcggc atccggtcag tggcagtgaa gggcgaacag      840

ttcctgatta accacaaacc gttctacttt actggctttg gtcgtcatga agatgcggac      900

ttgcgtggca aaggattcga taacgtgctg atggtgcacg accacgcatt aatggactgg      960

attggggcca actcctaccg tacctcgcat tacccttacg ctgaagagat gctcgactgg     1020

gcagatgaac atggcatcgt ggtgattgat gaaactgctg ctgtcggctt taacctctct     1080

ttaggcattg gtttcgaagc gggcaacaag ccgaaagaac tgtacagcga agaggcagtc     1140

aacggggaaa ctcagcaagc gcacttacag gcgattaaag agctgatagc gcgtgacaaa     1200

aaccacccaa gcgtggtgat gtggagtatt gccaacgaac cggatacccg tccgcaaggt     1260

gcacgggaat atttcgcgcc actggcggaa gcaacgcgta aactcgaccc gacgcgtccg     1320

atcacctgcg tcaatgtaat gttctgcgac gctcacaccg ataccatcag cgatctcttt     1380

gatgtgctgt gcctgaaccg ttattacgga tggtatgtcc aaagcggcga tttggaaacg     1440

gcagagaagg tactggaaaa agaacttctg gcctggcagg agaaactgca tcagccgatt     1500

atcatcaccg aatacggcgt ggatacgtta gccgggctgc actcaatgta caccgacatg     1560

tggagtgaag agtatcagtg tgcatggctg gatatgtatc accgcgtctt tgatcgcgtc     1620

agcgccgtcg tcggtgaaca ggtatggaat ttcgccgatt ttgcgacctc gcaaggcata     1680

ttgcgcgttg gcggtaacaa gaaagggatc ttcactcgcg accgcaaacc gaagtcggcg     1740

gcttttctgc tgcaaaaacg ctggactggc atgaacttcg gtgaaaaacc gcagcaggga     1800

ggcaaacaat ga                                                         1812


<210>  25
<211>  603
<212>  PRT
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(603)
<223>  gusA protein sequence

<400>  25

Met Leu Arg Pro Val Glu Thr Pro Thr Arg Glu Ile Lys Lys Leu Asp 
1               5                   10                  15      


Gly Leu Trp Ala Phe Ser Leu Asp Arg Glu Asn Cys Gly Ile Asp Gln 
            20                  25                  30          


Arg Trp Trp Glu Ser Ala Leu Gln Glu Ser Arg Ala Ile Ala Val Pro 
        35                  40                  45              


Gly Ser Phe Asn Asp Gln Phe Ala Asp Ala Asp Ile Arg Asn Tyr Ala 
    50                  55                  60                  


Gly Asn Val Trp Tyr Gln Arg Glu Val Phe Ile Pro Lys Gly Trp Ala 
65                  70                  75                  80  


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


Val Trp Val Asn Asn Gln Glu Val Met Glu His Gln Gly Gly Tyr Thr 
            100                 105                 110         


Pro Phe Glu Ala Asp Val Thr Pro Tyr Val Ile Ala Gly Lys Ser Val 
        115                 120                 125             


Arg Ile Thr Val Cys Val Asn Asn Glu Leu Asn Trp Gln Thr Ile Pro 
    130                 135                 140                 


Pro Gly Met Val Ile Thr Asp Glu Asn Gly Lys Lys Lys Gln Ser Tyr 
145                 150                 155                 160 


Phe His Asp Phe Phe Asn Tyr Ala Gly Ile His Arg Ser Val Met Leu 
                165                 170                 175     


Tyr Thr Thr Pro Asn Thr Trp Val Asp Asp Ile Thr Val Val Thr His 
            180                 185                 190         


Val Ala Gln Asp Cys Asn His Ala Ser Val Asp Trp Gln Val Val Ala 
        195                 200                 205             


Asn Gly Asp Val Ser Val Glu Leu Arg Asp Ala Asp Gln Gln Val Val 
    210                 215                 220                 


Ala Thr Gly Gln Gly Thr Ser Gly Thr Leu Gln Val Val Asn Pro His 
225                 230                 235                 240 


Leu Trp Gln Pro Gly Glu Gly Tyr Leu Tyr Glu Leu Cys Val Thr Ala 
                245                 250                 255     


Lys Ser Gln Thr Glu Cys Asp Ile Tyr Pro Leu Arg Val Gly Ile Arg 
            260                 265                 270         


Ser Val Ala Val Lys Gly Glu Gln Phe Leu Ile Asn His Lys Pro Phe 
        275                 280                 285             


Tyr Phe Thr Gly Phe Gly Arg His Glu Asp Ala Asp Leu Arg Gly Lys 
    290                 295                 300                 


Gly Phe Asp Asn Val Leu Met Val His Asp His Ala Leu Met Asp Trp 
305                 310                 315                 320 


Ile Gly Ala Asn Ser Tyr Arg Thr Ser His Tyr Pro Tyr Ala Glu Glu 
                325                 330                 335     


Met Leu Asp Trp Ala Asp Glu His Gly Ile Val Val Ile Asp Glu Thr 
            340                 345                 350         


Ala Ala Val Gly Phe Asn Leu Ser Leu Gly Ile Gly Phe Glu Ala Gly 
        355                 360                 365             


Asn Lys Pro Lys Glu Leu Tyr Ser Glu Glu Ala Val Asn Gly Glu Thr 
    370                 375                 380                 


Gln Gln Ala His Leu Gln Ala Ile Lys Glu Leu Ile Ala Arg Asp Lys 
385                 390                 395                 400 


Asn His Pro Ser Val Val Met Trp Ser Ile Ala Asn Glu Pro Asp Thr 
                405                 410                 415     


Arg Pro Gln Gly Ala Arg Glu Tyr Phe Ala Pro Leu Ala Glu Ala Thr 
            420                 425                 430         


Arg Lys Leu Asp Pro Thr Arg Pro Ile Thr Cys Val Asn Val Met Phe 
        435                 440                 445             


Cys Asp Ala His Thr Asp Thr Ile Ser Asp Leu Phe Asp Val Leu Cys 
    450                 455                 460                 


Leu Asn Arg Tyr Tyr Gly Trp Tyr Val Gln Ser Gly Asp Leu Glu Thr 
465                 470                 475                 480 


Ala Glu Lys Val Leu Glu Lys Glu Leu Leu Ala Trp Gln Glu Lys Leu 
                485                 490                 495     


His Gln Pro Ile Ile Ile Thr Glu Tyr Gly Val Asp Thr Leu Ala Gly 
            500                 505                 510         


Leu His Ser Met Tyr Thr Asp Met Trp Ser Glu Glu Tyr Gln Cys Ala 
        515                 520                 525             


Trp Leu Asp Met Tyr His Arg Val Phe Asp Arg Val Ser Ala Val Val 
    530                 535                 540                 


Gly Glu Gln Val Trp Asn Phe Ala Asp Phe Ala Thr Ser Gln Gly Ile 
545                 550                 555                 560 


Leu Arg Val Gly Gly Asn Lys Lys Gly Ile Phe Thr Arg Asp Arg Lys 
                565                 570                 575     


Pro Lys Ser Ala Ala Phe Leu Leu Gln Lys Arg Trp Thr Gly Met Asn 
            580                 585                 590         


Phe Gly Glu Lys Pro Gln Gln Gly Gly Lys Gln 
        595                 600             


<210>  26
<211>  122
<212>  DNA
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(122)
<223>  LacZ promoter

<400>  26
gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat       60

gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag      120

ct                                                                     122


<210>  27
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 23 sgRNA DNA sequence

<400>  27
agcggataac aatttcacac gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  28
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 24 sgRNA DNA sequence

<400>  28
cgtcgccacc aatccccata gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  29
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 64 sgRNA DNA sequence

<400>  29
cgttttacaa cgtcgtgact gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  30
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 65 sgRNA DNA sequence

<400>  30
ggccagtgaa tccgtaatca gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  31
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 66 sgRNA DNA sequence

<400>  31
cttcttccgc gtgcagcaga gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  32
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 67 sgRNA DNA sequence

<400>  32
ggcacatggc tgaatatcga gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  33
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 13 sgRNA DNA sequence

<400>  33
cggcctgtgg gcattcagtc gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  34
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 14 sgRNA DNA sequence

<400>  34
actgtggaat tgatcagcgt gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  35
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 15 sgRNA DNA sequence

<400>  35
agccgggcaa ttgctgtgcc gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  36
<211>  102
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Guide 16 sgRNA DNA sequence

<400>  36
tatcgtgctg cgtttcgatg gttttagagc tagaaatagc aagttaaaat aaggctagtc       60

cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tt                         102


<210>  37
<211>  1359
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dsDNA template for gusABC KO

<400>  37
ttatttccat ttctcttcca tgggtttctc acagataact gtgtgcaaca cagaattggt       60

taactaatca gattaaaggt tgaccagtat tattatctta atgaggagtc ccttatgtta      120

gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctta      180

gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa      240

ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg      300

atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca      360

ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct      420

tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc      480

gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc      540

ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc      600

gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg      660

ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc      720

atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac      780

caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat      840

caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc      900

aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg      960

aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg     1020

gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc     1080

gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc     1140

gccttctatc gccttcttga cgagttcttc tgagcgggac tctaaagcgc tctgaagttc     1200

ctatactttc tagagaatag gaacttcgga ataggaacta agtaaaaaat aacgccggag     1260

agaaaaatct ccggcgtttc agattgttga caaagtgcgc gttttttatg ccggatgcgg     1320

cgtaaacgcc ttatccagcc tacaaaaact cataaattc                            1359


<210>  38
<211>  1005
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: kanamycin resistance cassette

<400>  38
aagatcccct tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg       60

acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc      120

aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct      180

gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt      240

tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc      300

tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag      360

accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg      420

gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac      480

tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc      540

gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc      600

tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc      660

ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg      720

ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat      780

gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc      840

cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa      900

gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat      960

tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctga                     1005


<210>  39
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: FRT site 1

<400>  39
gaagttccta tactttctag agaataggaa cttcggaata ggaacttc                    48


<210>  40
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: FRT site 2

<400>  40
gaagttccta tactttctag agaataggaa cttcggaata ggaact                      46


<210>  41
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: homology region 1 GusA

<400>  41
ttatttccat ttctcttcca tgggtttctc acagataact gtgtgcaaca cagaattggt       60

taactaatca gattaaaggt tgaccagtat tattatctta atgaggagtc ccttatgtta      120


<210>  42
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: homology region 2 GusC

<400>  42
caatgaatca acaactctcc tggcgcacca tcgtcggcta cagcctcggt gacgtcgcca       60

ataacttcgc cttcgcaatg ggggcgctct tcctgttgag ttactacacc gacgtcgctg      120


<210>  43
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: gB primer 1

<400>  43
ttatttccat ttctcttcca tg                                                22


<210>  44
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: gB primer 2

<400>  44
gaatttatga gtttttgtag gc                                                22


<210>  45
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: homology region 1 GusA

<400>  45
caatgaatca acaactctcc tggcgcacca tcgtcggcta cagcctcggt gacgtcgcca       60

ataacttcgc cttcgcaatg ggggcgctct tcctgttgag ttactacacc gacgtcgctg      120


<210>  46
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: homology region 1 GusB

<400>  46
taacaagaaa gggatcttca ctcgcgaccg caaaccgaag tcggcggctt ttctgctgca       60

aaaacgctgg actggcatga acttcggtga aaaaccgcag cagggaggca aacaatgaat      120


<210>  47
<211>  1374
<212>  DNA
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(1374)
<223>  GusB DNA sequence

<400>  47
atgaatcaac aactctcctg gcgcaccatc gtcggctaca gcctcggtga cgtcgccaat       60

aacttcgcct tcgcaatggg ggcgctcttc ctgttgagtt actacaccga cgtcgctggc      120

gtcggtgccg ctgcggcggg caccatgctg ttactggtgc gggtattcga tgccttcgcc      180

gacgtctttg ccggacgagt ggtggacagt gtgaataccc gctggggaaa attccgcccg      240

tttttactct tcggtactgc gccgttaatg atcttcagcg tgctggtatt ctgggtgctg      300

accgactgga gccatggtag caaagtggtg tatgcatatt tgacctacat gggcctcggg      360

ctttgctaca gcctggtgaa tattccttat ggttcacttg ctaccgcgat gacccaacaa      420

ccacaatccc gcgcccgtct gggcgcggct cgtgggattg ccgcttcatt gacctttgtc      480

tgcctggcat ttctgatagg accgagcatt aagaactcca gcccggaaga gatggtgtcg      540

gtataccatt tctggacaat tgtgctggcg attgccggaa tggtgcttta cttcatctgc      600

ttcaaatcga cgcgtgagaa tgtggtacgt atcgttgcgc agccgtcatt gaatatcagt      660

ctgcaaaccc tgaaacggaa tcgcccgctg tttatgttgt gcatcggtgc gctgtgtgtg      720

ctgatttcga cctttgcggt cagcgcctcg tcgttgttct acgtgcgcta tgtgttaaat      780

gataccgggc tgttcactgt gctggtactg gtgcaaaacc tggttggtac tgtggcatcg      840

gcaccgctgg tgccggggat ggtcgcgagg atcggtaaaa agaatacctt cctgattggc      900

gctttgctgg gaacctgcgg ttatctgctg ttcttctggg tttccgtctg gtcactgccg      960

gtggcgttgg ttgcgttggc catcgcttca attggtcagg gcgttaccat gaccgtgatg     1020

tgggcgctgg aagctgatac cgtagaatac ggtgaatacc tgaccggcgt gcgaattgaa     1080

gggctcacct attcactatt ctcatttacc cgtaaatgcg gtcaggcaat cggaggttca     1140

attcctgcct ttattttggg gttaagcgga tatatcgcca atcaggtgca aacgccggaa     1200

gttattatgg gcatccgcac atcaattgcc ttagtacctt gcggatttat gctactggca     1260

ttcgttatta tctggtttta tccgctcacg gataaaaaat tcaaagaaat cgtggttgaa     1320

attgataatc gtaaaaaagt gcagcagcaa ttaatcagcg atatcactaa ttaa           1374


<210>  48
<211>  457
<212>  PRT
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(457)
<223>  GusB amino acid sequence

<400>  48

Met Asn Gln Gln Leu Ser Trp Arg Thr Ile Val Gly Tyr Ser Leu Gly 
1               5                   10                  15      


Asp Val Ala Asn Asn Phe Ala Phe Ala Met Gly Ala Leu Phe Leu Leu 
            20                  25                  30          


Ser Tyr Tyr Thr Asp Val Ala Gly Val Gly Ala Ala Ala Ala Gly Thr 
        35                  40                  45              


Met Leu Leu Leu Val Arg Val Phe Asp Ala Phe Ala Asp Val Phe Ala 
    50                  55                  60                  


Gly Arg Val Val Asp Ser Val Asn Thr Arg Trp Gly Lys Phe Arg Pro 
65                  70                  75                  80  


Phe Leu Leu Phe Gly Thr Ala Pro Leu Met Ile Phe Ser Val Leu Val 
                85                  90                  95      


Phe Trp Val Leu Thr Asp Trp Ser His Gly Ser Lys Val Val Tyr Ala 
            100                 105                 110         


Tyr Leu Thr Tyr Met Gly Leu Gly Leu Cys Tyr Ser Leu Val Asn Ile 
        115                 120                 125             


Pro Tyr Gly Ser Leu Ala Thr Ala Met Thr Gln Gln Pro Gln Ser Arg 
    130                 135                 140                 


Ala Arg Leu Gly Ala Ala Arg Gly Ile Ala Ala Ser Leu Thr Phe Val 
145                 150                 155                 160 


Cys Leu Ala Phe Leu Ile Gly Pro Ser Ile Lys Asn Ser Ser Pro Glu 
                165                 170                 175     


Glu Met Val Ser Val Tyr His Phe Trp Thr Ile Val Leu Ala Ile Ala 
            180                 185                 190         


Gly Met Val Leu Tyr Phe Ile Cys Phe Lys Ser Thr Arg Glu Asn Val 
        195                 200                 205             


Val Arg Ile Val Ala Gln Pro Ser Leu Asn Ile Ser Leu Gln Thr Leu 
    210                 215                 220                 


Lys Arg Asn Arg Pro Leu Phe Met Leu Cys Ile Gly Ala Leu Cys Val 
225                 230                 235                 240 


Leu Ile Ser Thr Phe Ala Val Ser Ala Ser Ser Leu Phe Tyr Val Arg 
                245                 250                 255     


Tyr Val Leu Asn Asp Thr Gly Leu Phe Thr Val Leu Val Leu Val Gln 
            260                 265                 270         


Asn Leu Val Gly Thr Val Ala Ser Ala Pro Leu Val Pro Gly Met Val 
        275                 280                 285             


Ala Arg Ile Gly Lys Lys Asn Thr Phe Leu Ile Gly Ala Leu Leu Gly 
    290                 295                 300                 


Thr Cys Gly Tyr Leu Leu Phe Phe Trp Val Ser Val Trp Ser Leu Pro 
305                 310                 315                 320 


Val Ala Leu Val Ala Leu Ala Ile Ala Ser Ile Gly Gln Gly Val Thr 
                325                 330                 335     


Met Thr Val Met Trp Ala Leu Glu Ala Asp Thr Val Glu Tyr Gly Glu 
            340                 345                 350         


Tyr Leu Thr Gly Val Arg Ile Glu Gly Leu Thr Tyr Ser Leu Phe Ser 
        355                 360                 365             


Phe Thr Arg Lys Cys Gly Gln Ala Ile Gly Gly Ser Ile Pro Ala Phe 
    370                 375                 380                 


Ile Leu Gly Leu Ser Gly Tyr Ile Ala Asn Gln Val Gln Thr Pro Glu 
385                 390                 395                 400 


Val Ile Met Gly Ile Arg Thr Ser Ile Ala Leu Val Pro Cys Gly Phe 
                405                 410                 415     


Met Leu Leu Ala Phe Val Ile Ile Trp Phe Tyr Pro Leu Thr Asp Lys 
            420                 425                 430         


Lys Phe Lys Glu Ile Val Val Glu Ile Asp Asn Arg Lys Lys Val Gln 
        435                 440                 445             


Gln Gln Leu Ile Ser Asp Ile Thr Asn 
    450                 455         


<210>  49
<211>  1266
<212>  DNA
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(1266)
<223>  GusC DNA sequence

<400>  49
atgagaaaaa tagtggccat ggccgttatt tgcctgacgg ctgcctctgg ccttacctct       60

gcttatgcgg cgcaactggc tgacgatgaa gcgggactac gcatcagact gaaaaacgaa      120

ttgcgcaggg cggataagcc cagtgctggc gcgggaagag atatttacgc atgggtacag      180

ggaggattgc tcgatttcaa tagtggttat tattccaata ttattggcgt tgaaggcggg      240

gcgtattatg tttataaatt aggtgctcgt gctgatatga gtacccggtg gtatcttgat      300

ggtgataaaa gttttggctt tgccctgggg gcagtaaaaa taaaacccag tgaaaatagc      360

ctgcttaaat taggtcgctt cgggacggat tatagttatg gtagcttacc ttatcgtatt      420

ccgttaatgg ctggcagttc gcaacgtaca ttaccgacag tttctgaagg agcattaggt      480

tattgggctt taacaccaaa tattgatctg tggggaatgt ggcgttcacg agtattttta      540

tggactgatt caacaaccgg tattcgtgat gaaggggtgt ataacagcca gacgggaaaa      600

tacgataaac atcgcgcacg ttctttttta gccgccagtt ggcatgatga taccagtcgc      660

tattctctgg gggcatcggt acagaaagat gtttccaatc agatacaaag tattctcgag      720

aaaagcatac cgctcgaccc gaattatacg ttgaaagggg agttgctcgg cttttacgcg      780

cagctcgaag gtttaagtcg taataccagc cagcccaatg aaacggcgtt ggttagtgga      840

caattgacct ggaatgcgcc gtggggaagt gtatttggca gtggtggtta tttgcgccat      900

gcaatgaatg gtgccgtggt ggataccgac attggctatc ccttttcatt aagtcttgat      960

cgtaaccgtg aaggaatgca gtcctggcaa ttgggcgtca actatcgttt aacgccgcaa     1020

tttacgctga catttgcacc gattgtgact cgcggctatg aatccagtaa acgagatgtg     1080

cggattgaag gcacgggtat cttaggtggt atgaactatc gggtcagcga agggccgtta     1140

caagggatga atttctttct tgctgccgat aaagggcggg aaaagcgcga tggcagtacg     1200

ctgggcgatc gcctgaatta ctgggatgtg aaaatgagta ttcagtatga ctttatgctg     1260

aagtaa                                                                1266


<210>  50
<211>  421
<212>  PRT
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (1)..(421)
<223>  GusC amino acid sequence

<400>  50

Met Arg Lys Ile Val Ala Met Ala Val Ile Cys Leu Thr Ala Ala Ser 
1               5                   10                  15      


Gly Leu Thr Ser Ala Tyr Ala Ala Gln Leu Ala Asp Asp Glu Ala Gly 
            20                  25                  30          


Leu Arg Ile Arg Leu Lys Asn Glu Leu Arg Arg Ala Asp Lys Pro Ser 
        35                  40                  45              


Ala Gly Ala Gly Arg Asp Ile Tyr Ala Trp Val Gln Gly Gly Leu Leu 
    50                  55                  60                  


Asp Phe Asn Ser Gly Tyr Tyr Ser Asn Ile Ile Gly Val Glu Gly Gly 
65                  70                  75                  80  


Ala Tyr Tyr Val Tyr Lys Leu Gly Ala Arg Ala Asp Met Ser Thr Arg 
                85                  90                  95      


Trp Tyr Leu Asp Gly Asp Lys Ser Phe Gly Phe Ala Leu Gly Ala Val 
            100                 105                 110         


Lys Ile Lys Pro Ser Glu Asn Ser Leu Leu Lys Leu Gly Arg Phe Gly 
        115                 120                 125             


Thr Asp Tyr Ser Tyr Gly Ser Leu Pro Tyr Arg Ile Pro Leu Met Ala 
    130                 135                 140                 


Gly Ser Ser Gln Arg Thr Leu Pro Thr Val Ser Glu Gly Ala Leu Gly 
145                 150                 155                 160 


Tyr Trp Ala Leu Thr Pro Asn Ile Asp Leu Trp Gly Met Trp Arg Ser 
                165                 170                 175     


Arg Val Phe Leu Trp Thr Asp Ser Thr Thr Gly Ile Arg Asp Glu Gly 
            180                 185                 190         


Val Tyr Asn Ser Gln Thr Gly Lys Tyr Asp Lys His Arg Ala Arg Ser 
        195                 200                 205             


Phe Leu Ala Ala Ser Trp His Asp Asp Thr Ser Arg Tyr Ser Leu Gly 
    210                 215                 220                 


Ala Ser Val Gln Lys Asp Val Ser Asn Gln Ile Gln Ser Ile Leu Glu 
225                 230                 235                 240 


Lys Ser Ile Pro Leu Asp Pro Asn Tyr Thr Leu Lys Gly Glu Leu Leu 
                245                 250                 255     


Gly Phe Tyr Ala Gln Leu Glu Gly Leu Ser Arg Asn Thr Ser Gln Pro 
            260                 265                 270         


Asn Glu Thr Ala Leu Val Ser Gly Gln Leu Thr Trp Asn Ala Pro Trp 
        275                 280                 285             


Gly Ser Val Phe Gly Ser Gly Gly Tyr Leu Arg His Ala Met Asn Gly 
    290                 295                 300                 


Ala Val Val Asp Thr Asp Ile Gly Tyr Pro Phe Ser Leu Ser Leu Asp 
305                 310                 315                 320 


Arg Asn Arg Glu Gly Met Gln Ser Trp Gln Leu Gly Val Asn Tyr Arg 
                325                 330                 335     


Leu Thr Pro Gln Phe Thr Leu Thr Phe Ala Pro Ile Val Thr Arg Gly 
            340                 345                 350         


Tyr Glu Ser Ser Lys Arg Asp Val Arg Ile Glu Gly Thr Gly Ile Leu 
        355                 360                 365             


Gly Gly Met Asn Tyr Arg Val Ser Glu Gly Pro Leu Gln Gly Met Asn 
    370                 375                 380                 


Phe Phe Leu Ala Ala Asp Lys Gly Arg Glu Lys Arg Asp Gly Ser Thr 
385                 390                 395                 400 


Leu Gly Asp Arg Leu Asn Tyr Trp Asp Val Lys Met Ser Ile Gln Tyr 
                405                 410                 415     


Asp Phe Met Leu Lys 
            420     


