multilocus sequence typing of bacteria: a genotypic tool for strain selection aundy kumar, *jos m....

Multilocus Sequence Typing of bacteria:a genotypic tool for strain selection

Aundy Kumar, *Jos M. Raaijmakers, Santhosh J Eapen

Indian Institute of Spices Research, Calicut

___________________________________________________________________**Laboratory of Phytopathology, Wageningen University,

The Netherlands

Image courtesy: pseudomonas.com

Tools and web resources at your door step!

Advances in sequencing methodsEver reducing cost of sequencingFreely available analytical toolsGood databases with constant updates

Full genome information on 1400 bacteria

http://pubmlst.org/http://www.mlst.net/http://www.pasteur.fr/recherche/genopole/PF8/mlst/

PseudomonasAcinetobacterKlebsiellaStenotrophomonasEnterobacterBacillus Burkholderia

MLST scheme has been developed for ~40 bacteria so far

Frequently encountered ones

What is MLST?

A nucleotide sequence based approach to uncover

allelic variants in conserved genes (house keeping)

for the purpose of characterizing, subtyping, and

classifying members of bacterial populations.

Proposed in 1998 using the human pathogen

Neisseria meningitidis as an example

A portable, universal, and definitive method for

characterizing bacteria

MLST – genes used

Pseudomonas aerugenosaacsA acetyl coenzyme AaroE Shikimate dehydrogenase

guaA GMP synthasemutL DNA mismatch repair proteinnuoD NADH dehydrogenase IppsA Phosphoenol pyruvate

synthasetrpE Anthralite synthetase

Ralstonia solanacearum

Glutamate dehydrogenase oxidoreductase

gdhA

Adenylate kinase adk

DNA gyrase subunit B gyrB

Glyceraldehyde 3-phosphate dehydrogeanase oxidoreductase

gapA

Phosphoenol pyruvate synthase

ppsA

Flagellin protein fli C

Regulatory transcription regulator

hrpB

Endoglucanase precursor

egl

PCR amplification of selected genes

Elution & purification of PCR product

Sequencing of PCR product

Sequence assembly (DNA Baser)

Blast the assembled sequence in database www.PAMDB.org

Assigning allele numbers for each gene

Unique allelic profile is called the Sequence Type(ST)

PROTOCOL

Each sequence for a given locus is screened for identity (not for similarity) with already known sequences for that locus. If the sequence is different, it is considered to be a new allele and is assigned a unique (arbitrary) allele number

Case study of Multilocus Sequence Typing of plant associated Pseudomonas aeruginosa

In Spice crops

Projects on bacterial endophytes in black pepper/ginger against major soil borne pathogens

…has yielded several diverse group of bacterial species

The best among the lot is

Pseudomonas aeruginosa

Aravind et al Letters in Applied Microbiology, 2009

Strain Source Year of collection

Isolated by

Pa-GEB9 Ginger, Calicut (against Pythium)

2007 Aundy Kumar

Pa-IISR51 Ginger, Karnataka (against Pythium)

1998 Aundy Kumar

Pa-BP35 Black pepper, Kerala (against Phytophthora)

2006 Aundy Kumar

Pa-IISR13 Black pepper, Andhra Pradesh (against Phytophthora)

1998 Y.R.Sarma

Pa-IISR6 Black pepper, Kerala (against Phytophthora)

2000 Y.R.Sarma

Pa-IISR853

Chromolaena odorata (against nematodes)

2005 S.J.Eapen

PAO1- Type strain

Human burn patient, Australia

1950s Bruce Holloway

Selected from several collection in various projects during different points

Some of them are endophytes in plants

…b

ackg

rou

nd

0

10

20

30

40

50

60

70

80

90

100

10 9 8 7 6 5 4 3 2 0

Bacterial concentration in suspension (log CFU per mL)

Lesi

on in

hibi

tion

(%)

0

1

2

3

4

5

6

7

8

Bacterial population in shoot

as endophyte (log CFU per

gram of shoot)

Inhibition of P. capsici induced les ion (%) by BP35 strain

Endophytic population (Log CFU g-1)

DN

A of

BP3

5

DNA extracted from bacterized stem is used as template

578bp

Direct evidence for protection of black pepper

by specific density of P. aeruginosa BP35 cells

PhytophthoraSuppression

90-100%

BP35Density in Suspension

8-10 log units

EndogenousPopulation of

BP354.5-6.5 log

units

Phenotype- functional

Leaf

Roots

StemLocalization of BP35::gfp2 in tomato

Accumulation of bacterial cells

Tracking of bacterium PaBP35

15 Days Post Bacterization in Five weeks old tomato cultivar, Moneymaker

Tissues fixed in 4% paraformaldehyde

Localized in aerial plant parts

The bacterium was tagged with gpf using Tn7 system

Phenotype- functional

Few of them are in field evaluation stage for

1. Phytophthora suppression2. Radopholus similis suppression

Single or Two nodal cutting are Bioprimed

Bacterized shoots

March 2010

July 2010

August 2010

Metalaxyl

Collectively they may be classified as Plant Associated Pseudomonas aeruginosa

Pseudomonas aeruginosa-

Type strain for Pseudomonas

Most intensively studied bacterium

Opportunistic human pathogen

Full genome published in Nature

MLST analysis

Analysis of housekeeping genes that are scattered in the genome

www.pseudomonas.com

Gene Amplicon (bp)

Position in the PAO1 genome

Sequence for MLST analysis (bp)

Coordinates in the PAO1 genome

No of alleles documented so far

acsA -(acetyl coenzyme A)

842 969670 390 970604- 971033

145

aroE-(Shikimate dehydrogenase)

1042 26711 495 26883-27380

146

guaA-(GMP synthase) 940 4227237 372 4226567-4226155 115

mutL-(DNA mismatch repair protein)

940 5551681 441 5551179- 5550698

124

nuoD- (NADH dehydrogenase I)

1042 2983963 366 2984570- 2984955

86

ppsA- (Phosphoenol pyruvate synthase)

989 1914037 369 1914994- 1915403

97

trpE- Anthralite synthetase

811 670980 441 671811- 672293

129

Choice of genes for MLST analysis

Genes were amplified using specific primers

The amplicon were eluted using gel elution kit (Sigma Aldrich)

Sequenced (Both forward and reverse run)

M: DNA ladder 50bp

aro

E

acsA

gu

aA

mu

tL

nu

oD

pp

sA

trp

E

BP3

5

PAO

1

acs

A a

roE

guaA

mutL

nuo

D ppsA

trpE

acs

A a

roE

guaA

mutL

nuo

D ppsA

trpE

1000bp

Out group from clinical origin

Gene amplification, isolation, sequencing and sequence analysis

aro

E

acsA

gu

aA

m

utL

n

uoD

p

psA

trp

E aro

E

acsA

gu

aA

m

utL

n

uoD

p

psA

trp

E

BP35 IISR 853 GEB 9

aro

E

acsA

gu

aA

mu

tL

nu

oD

p

psA

trp

Earo

E

acsA

gu

aA

m

utL

n

uoD

p

psA

trp

E aro

E

acsA

gu

aA

mu

tL

nu

oD

p

psA

trp

E

aro

E

acsA

gu

aA

m

utL

n

uoD

p

psA

trp

E

IISR 6 IISR 51 IISR 13

All seven genes were

isolated, sequenced

and analysed

The sequences were assembled and analysed using Vector NTiSequence viewer/DNA baser

Alleles were determined using pubmlst.org

String of alleles (Sequence Type) were analysed using eBurst algorithm (eBurst V3) or profuse algorithm (goeBurst.org)

>acsA-IISR6TATAACTGTNGGNTTNAACGCGCTGACCCACGAGCGGGTCTTCGACTACCGTCCGGGCGAAGTCTACTGGTGCACCCGCCGGACATCGGCTGGGTCACCGGCCACACCTACATCGTCTATGGCCCGCTGGCCAACGGCGCCACCACCATTCTGTTCGAGGGCGTGCCGAACTACCCCGACGTGACCCGCGTGGCGAAAATCATCGACAAGCACAAGGTCAACATCCTCTACACCGCGCCGACCGCGATCCGCGCGATGATGGCCGAAGGCAAGGCGGCGGTGGCCGGTGCCGACGGTTCCAGCCTGCGTCTGCTCGGTTCGGTGGGCGAGCCGATCAACCCGGAAGCCTGGCAGTGGTACTACGAGACCGTCGGCCAGTCGCGCTGCCCGATCGTCGACACCTGGTGGCAGACCGAGACCGGCGCCTGCCTGATGACCCCGCTGCCGGGCGCCCACGCGATGAAGCCGGGCTCTGCAGCCAAGCCGTTCTTCGGCGTGGTACCGGCGCTGGTGGACAACCTCGGCAACCTGATCGAGGGCGCCGCCGAGGGCAACCTGGTGATCCTCGACTCCTGGCCGGGCCAGGCGCGGACCCTGTTCGGCGACCATGACCGCTTCGTCGACACCTACTTCAAGACCTTCAAGGGCATGTACTTCACCGGCGACGGCGCGCGCCGCGACGAGGACGGCTACTACTGGATCACCGGGCGGGTCGACGACGTGCTCAACGTCTCCGGCCACCGCATGGGCACCGCCGAGGTGGAAAGCGCGATGGTCGCCCACCCGAAGGTCGCCGAGGCGGCGGTGGTCGGCATGCAGCACGACATCAAGGGGAGCG

Allele: 15

>aroE-IISR6CATGTCAGATGGAAACCGATTGAGACGTACGTCTCAAATGCTGCCTGACAACTCAGACAGTTGCCAATTGCCGGCGCAGCGTCTCCAACACTGGCGCCGAGGCAGGACGCACGCCACGCCAGAGGAAGAATGCCTCGGCGGCCTGCTCCACCAGCATGCCCAGGCCATCCAGGGTACGCGCCGCACCGCGTTCGGCGGCCCAGCGGTTGAAGGCGGTCGGTTCCTTGGCATACATCATGTCGTAGCAAACGGTACGGCCGGGCTCGATCACGCTCTGCGCCAGCGGCGGCACGTCGCCGGCAAGACTGGCCGAGGTGCCGTTGACGACCAGGTCGAAAGGCCCTTCGACCTCGGCGAAACCGCAGCCGCGCACCGCGCCGAGATCGGCGAACCGCTCGGCCAGATCCACGGCCTTCCGCGCCGTGCGGTTGGCGATCAGCAGCTCCGCCGGGCACTCGCCGAGGAAGGGTTCGAGCACCCCGCGCACCGCACCGCCGGCGCCGAGCAGGAGAACCCGCTTGCCGCGCAGCTCGACCCCGGCGTTCGCCGTCAGGTCCCGCAGCAAGCCCGCGCCGTCGGTGTTGTCGCCGCGCAGGCGACCGTCGGCCAGGCGGATGAGGGTGTTCACCGCCCCGGCCCGGGTGGCCCGCTCGCTCAATTCGTCCACCAGACGATAGGCCTCTTCCTTGAACGGCACGGTGACATTGGCGCCCTTGCCCTGCTCGAAGAAGCGCCGGGCGAACCCGGGGAAATCGTCCAGCGGCGCCAGCTGCGCGTCATAGACCAGCGCCTCGCCGGTCTGCTCGGCGAACAGGCGGTGGATCAGCGGCGACTTGCTGTGGCCGATGGGGTTGCCGAATACGCAATAGCGGTCCATGGGTCGTTCCTTTGCCGGGGTGGGACGGGACGAGCGGACTCGTCCCCTGCGGGCGGGTCAGGGCTGGCCGGCCAGCCAATCGCGGGTCGGCGAGGAAATACTCGGTCAGGCGCGCTTCCTCGCTGC

Allele: 121

>guaA-IISR6ANTNGNGGNCGNAACCNAANGNTGCCGCGGCCTGNCTTGGTGTTGGGTGACTTCCGGGGGGAATTCCAGCCGTTGNTGGCGCGGGAATTCGTNCCTTNAAGCGGTGGTGAGCCACGGCGACAAGGTCACCGAGATGCCGGCCGCGCTTCCACATCCTGGCCAGCACCCCGAGCTGCCCGATCGCCGCCATGGCAGACGATGCCCGCGCCTACTACGGCGTGCAATTCCACCCGGAAGTCACCCACACCAAGCAGGGCCTGCGCATTCTCTCGCGCTTCGTCCTCGACATCTGCGGTTGCGCCGCGCTGTGGACCCCCTCGAACATCGTCGACGACGCCATCGCCACCGTGCGCGCCCAGGTCGGTTCCTCCAAGGTCCTGCTAGGCCTCTCCGGCGGCGTGGACTCCTCGGTGGTCGCCGCGCTGCTGCACAAGGCCATCGGCGACCAACTGACCTGCGTGTTCGTCGACAACGGCCTGCTGCGCCTGCACGAAGGCGACCAGGTGATGGCCATGTTCGCCGAGAACATGGGCGTGAAGGTGATCCGCGCCAACGCCGAGGACAAGTTCCTCGGCCGCCTGGCCGGCGTCGCCGACCCGGAAGAGAAGCGCAAGATCATCGGCCGCACCTTCATCGAAGTCTTCGACGAAGAAGCCACCAAGCTGCAGGACGTGAAGTTCCTCGCCCAGGGCACCATCTACCCCGACGTGATCGAGTCGGCCGGCGCCAAGACCGGCAAGGCCCACGTGATCAAGTCGCACCACAACGTCGGCGGCCTGCCGGAGGACATGCAGTTCGAACTGGTCGAGCCGCTGCGCGAACTGTTCAAGGACGAAGTGCGCAAGATCGGCCTGGAGCTGGGCCTGCCCTACGACATGGTCTACCGCCACCCGTTCCCCGGCCCGGGCCTGGGCGTGCGCATCCTCGGCGAGGTGAAGAAAGAGTACGCCGACCTGCTGCGCCAGGCCGACCACATCTTCATCGAGGAACTGCGCGCCTTCGACTGTACCACAGACAGCCCGGGCGGTTCGATACANTGCNNCGCCTACNACGGCNGGCAATTCCGCCCNGAAGTCACTCACACAAGCANNNNGGCCANNCTCTGCCNGTTCCN

Allele: 36

>mutL-IISR6GGCGGTCGCGAGTCATGGTNGCGCGNACCGAACTAATCGAGCCAGCGCTCGCCACGGAGCCGNGGCTCATCCACCCGCCTTCAGAGANATGCGTCGCGTGAGTGGTCGAGCGTCCCGCCTCGGTGGCCAAGGAACTCTGGAAAACAGCTTGACGCCGGTTCCCGGGCGCATTGATGTGGAGGTCGAGCAGGGCGGCATCAAGTTGCTGCGAGTGCGCGACGACGGTCGCGGCATCCCCGCCGACGACCTGCCGCTGGCCCTGGCTCGCCACGCCACCAGCAAGATCCGCGAGCTGGAAGACCTGGAGCGGGTGATGAGCCTCGGCTTCCGTGGCGAGGCGCTGGCCTCGATCAGTTCGGTAGCGCGCCTGACCATGACCTCGCGTACCGCCGACGCCGGCGAAGCCTGGCAGGTGGAAACCGAGGGCCGTGACATGCAGCCGCGGGTACAGCCGGCGGCGCACCCGGTGGGGACCAGCGTCGAGGTTCGCGACCTGTTCTTCAACACCCCGGCGCGGCGCAAGTTCCTGCGCGCCGAGAAGACCGAGTTCGACCATCTGCAGGAAGTCATCAAGCGCCTGGCGCTGGCCCGTTTCGACGTGGCTTTCCACCTGCGCCACAACGGCAAGACCATCTTCGCCCTGCACGAGGCGCGAGACGAGCTGGCCCGCGCGCGCCGGGTCGGCGCGGTGTGCGGCCAGGCATTCCTCGAGCAGGCGCTGCCGATCGAGGTCGAGCGCAACGGCCTGCACCTGTGGGGCTGGGTCGGCTTGCCGACCTTCTCCCGCAGCCAGCCGGACCTGCAGTACTTCTATGTGAACGGGCGCATGGTGCGCGACAAGCTGGTCGCCCACGCGGTGCGCCAGGCTTATCGCGACGTGCTGTACAACGGCCGGCATCCGACCTTCGTGCTGTTCTTCGAAGTCGATCCGGCGGTTGGTGGACGTCANNTGCACCCGAACNGCACGAGTTCGCTTNNNGACAGCCGGATGNCCATGACCTTCCCTCTNNNGAGANACCCNNNGA

>nuoD-IISR6NTNNNGCCCACGCCAACTGGTACGAGCGCGAAGTGTGGGACATGTACGGCATCACCTTTCACCGGCCACCCGCACCTGACCCGCATGCTGATGCCGCCGACCTGGCAGGGCCATCCGCTGCGCAAGGACTACCCGGCGCGCGCCACCGAGTTCGATCCCTACTCCCTGTCCGCCGCCAAGCAGGACCTGGAGCAGGAAGCCCTGCGCTTCAAGCCGGAAGACTGGGGCATGAAGCGCCACGGCGAGAACGAGGACTACATGTTCCTCAACCTCGGCCCGAACCACCCGTCCGCCCACGGCGCGTTCCGCATCATCCTGCAACTGGACGGCGAGGAGATCATCGACTGCGTCCCGGAGATCGGCTACCACCACCGCGGCGCCGAGAAGATGGCCGAGCGCCAGTCCTGGCACAGTTTCATCCCCTACACCGACCGCATCGACTACCTCGGCGGGGTGATGAACAACCTGCCCTACGTACTCTCGGTGGAGAAGCTCGCCGGGATCAAGGTGCCCCAGCGGGTCGACGTGATCCGGATCATGATGGCGGAGTTCTTCCGTATCCTGAACCACCTGCTGTACCTGGGCACCTATATCCAGGACGTCGGCGCCATGACCCCGGTGTTCTTCACCTTCACCGACCGCCAGCGCGCCTACAAGGTGGTCGAGGCCATCACCGGCTTCCGCCTGCACCCGGCCTGGTACCGCATCGGCGGCGTCGCCCACGACCTGCCGCGCGGCTGGGACAAGCTGGTCCGCGAGTTCCTCGACTGGATGCCCAAGCGCCTCGACGAGTACGAGACCGCGGCCCTGAAGAACAGCATCTTGCGCGGCCGGACCATCGGCGTCGCCCAGTACAACACCAAGGAAGCCCTCGAGTGGGGCACCACCGGCGCCGGCCTGCGCGCCACCGGTTGCGACTTCGACCTGCGCAAGGCCCGCCCCTACTCCGGCTACGAGAACTTCGAGTTCGAGGTACCGCTGGCCCACAACGGCGACGCCTACGACCGCTGCATGGT

Allele: 4

>ppsA-IISR6AGATGCTTANGGAGCATAAGCTNTNGNAATCCTGGNNNCCCAANCCTCCTCCCTTCTATTNNNNGACGTCGATCGTNCGATCGCGCCNNTCGCCTGAGCGACGCCGAGGTCACCGAAGCTGGCCAAGCAGGCCATGATCATCGAGAAGCACTATGGCCGCCCGATGGACATCGAATGGGCCAAGGACGGTGACGACGGCAAGCTGTACATCGTCCAGGCACGCCCGGAAACCGTGAAGAGCCGCGCCAGCGCCACGGTCATGGAGCGCTACCTGCTGAAAGAGAAGGGGACCGTCCTGGTGGAAGGGCGTGCCATCGGCCAGCGCATCGGTGCCGGTCCGGTCAAGGTGATCAACGACGTGTCGGAAATGGACAAGGTCCAACCGGGTGACGTCCTGGTCTCCGACATGACCGACCCGGACTGGGAGCCAGTGATGAAGCGCGCCAGCGCCATCGTCACCAACCGCGGCGGGCGCACCTGCCACGCGGCGATCATCGCTCGCGAACTGGGCATCCCGGCGGTGGTCGGTTGCGGCAACGCCACCCAGATCCTGCAGGATGGCCAGGGGGTGACCGTTTCCTGTGCCGAAGGCGATACCGGCTTCATCTTCGAAGGCGAACTCGGTTTCGATGTGCGCAAGAACTCGGTCGACGCCATGCCCGACCTTCCGTTCAAGATCATGATGAACGTCGGCAATCCCGATCGCGCTTTCGATTTCGCCCAGTTGCCGAACGAAGGCGTGGGCCTGGCCCGCCTCGAATTCATCATCAACCGCATGATCGGCGTGCATCCCAAGGCGTTGCTGAACTTCGCCGGCCTGCCGGCGGACATCAAGGAAAGCGTGGAGAAGCGCATCGCCGGCTATCCCGATCCGGTCGGCTTCTACGTCGAGAAGCTGGTGGAGGGCATCAGCACCCTGGCCGCGGCGTTCTGGCCGAAGAACGGTCATCGTGCGCCTGTCCGACTTCAAGTCCAATGAGTACGCCACCTGATCGGCGGCAGCTCTACGAGCAGGAGAAACATAACNCTCCACCAATGAANANANNGACNANN

Allele: 15

>trpE-IISR6ATCNNNTATTCAAGNCTCAATCGTTCAAGNACGATCCAGCNACAGCGNNNNNNGCNNNGCTTCCNCCTAATCNCGNNAAATNNGNNNNGCGCCNNCAGCNGNNNNGGCGCGNGCACGGCTGAAGGCAATGAACGCATGCATCGATCGCGTATCAAACGTAACTGCACTGCATGCGGGGGCGGATCGTGCCACCTCCGTCCCGGCGGCTTGTGATGGTTTCTTCCCACTCCAGCGCCGGCACCGAGTCGGCAACGATACCGCCGCCGGCCTGCACGTGGAGTTCACCGTTCTTGATCACCGCGGTGCGGATGGCAATGGCGGTGTCCATGTTGCCGTTCCATGCCAGGTAGCCGACCGCGCCGCCGTAGACTCCACGCTTGACCGGCTCCAGCTCGTCGATGATCTCCATGGCGCGGATCTTCGGCGCGCCGGACAGCGTACCCGCCGGCAGGATCGCCCGCAGCGCGTCCATCGCGCTGAGCCCCTCGCGCAATTGCCCGGTGACGTTGGACACGATGTGCATGACGTTGGAGTAACGTTCGATCACCATTTTTTCGGTGACCTTCACCGCGCCGATATCGGACACCCGCCCCACGTCGTTGCGCCCCAGGTCGATCAGCATCAGGTGCTCGGCGATCTCCTTGGCGTCCGACAGCAGATCCTGCTCCAGCGCCAGGTCGGCCTCTTCGTTGATCCCGCGCGGACGGGTACCGGCGATCGGGCGCACCGTCACCAGGCCATCCTCGACCCGTACCAGCACCTCCGGCGAGCTGCCCACGACATGGAAGTCGCCGAAGTTGAAGAAGTACATGTAGGGCGTCGGATTGAAACAGCGCAGCGCGCGGTACAGGTCGATGGGCGCCGCCTTGAATTCGATGGACATGCGCTGCGACGGCACCACCTGCATGCAGTCGCCGGCCAGGATGTAGTCCTTGATCCTTCCTACCGCGTCTTCATAGTCCTCGCGGGTGAAGCTGGCACGAAACGCCAGCTCACGCCTTGGGGNCCCGCACANAGTGGGGGCGGGTGCCCCATCTGCGTGCCGTGAAGTTCACCGTTATAGATGANCGNTGTACGGATAGTCNTAGCTGTNTCATGTTGCCTTCAATGTCACGTGCAATCGCGCGCGTAGACCNCTCGACGGACGCCTCTCATGATATCCTGGCGGCAGTACGCTGCGCAGCGCANNGNAAGTNGNNNCATGGCTNGCCTCGGCAATGTCGGATCAG

Allele: 8

P. aeruginosa IISR6

Allele: 5

>acsA-IISR51CNNCAGGGGTTCTTGANTACCGTCCGGGCGAGTCTACTGGTGCACCGCCGACATCGGCTGGGTCACCGGCCACACCTACATCGTCTATGGCCCGCTGGCCAACGGCGCCACCACCATTCTGTTCGAGGGCGTGCCGAACTACCCCGACGTGACCCGCGTGGCGAAAATCATCGACAAGCACAAGGTCAACATCCTCTACACCGCGCCGACCGCGATCCGCGCGATGATGGCCGAAGGCAAGGCGGCGGTGGCCGGTGCCGACGGTTCCAGCCTGCGTCTGCTCGGTTCGGTGGGCGAGCCGATCAACCCGGAAGCCTGGCAGTGGTACTACGAGACCGTCGGCCAGTCGCGCTGCCCGATCGTCGACACCTGGTGGCACACCGAGACCGGCGCCTGCCTGATGACCCCGCTGCCGGGCGCCCACGCGATGAAGCCGGGCTCTGCAGCCAAGCCGTTCTTCGGCGTGGTACCGGCGCTGGTGGACAACCTCGGCAACCTGATCGAGGGCGCCGCCGAGGGCAACCTGGTGATCCTCGACTCCTGGCCGGGCCAGGCGCGGACCCTGTTCGGCGACCATGACCGCTTCGTCGACACCTACTTCAAGACCTTCAAGGGCATGTACTTCACCGGCGACGGCGCGCGCCGCGACGAGGACGGCTACTACTGGATCACCGGGCGGGTCGACGACGTGCTCAACGTCTCCGGCCACCGCATGGGCACCGCCGAGGTGGAAAGCGCGATGGTCGCCCACCCGAAGGTCGCCGAGGCGGCGGTGGTCGGCATGCAGCACGACATCAAGGGGCAGCA

Allele: 15

>aroE-IISR51TGNNTTNGNGGACGGATTTTGAGACGTACGTCTCAAATGCTGCCTGACAACTCAGACAGTTGCCAATTGCCGGCGCAGCGTCTCCAACACTGGCGCCGAGGCAGGACGCACGCCACGCCAGAGGAAGAATGCCTCGGCGGCCTGCTCCACCAGCATGCCCAGGCCATCCAGGGTACGCGCCGCACCGCGTTCGGCGGCCCAGCGGTTGAAGGCGGTCGGTTCCTTGGCATACATCATGTCGTAGCAAACGGTACGGCCGGGCTCGATCACGCTCTGCGCCAGCGGCGGCACGTCGCCGGCAAGACTGGCCGAGGTGCCGTTGACGACCAGGTCGAAAGGCCCTTCGACCTCGGCGAAACCGCAGCCGCGCACCGCGCCGAGATCGGCGAACCGCTCGGCCAGATCCACGGCCTTCCGCGCCGTGCGGTTGGCGATCAGCAGCTCCGCCGGGCACTCGCCGAGGAAGGGTTCGAGCACCCCGCGCACCGCACCGCCGGCGCCGAGCAGGAGAACCCGCTTGCCGCGCAGCTCGACCCCGGCGTTCGCCGTCAGGTCCCGCAGCAAGCCCGCGCCGTCGGTGTTGTCGCCGCGCAGGCGACCGTCGGCCAGGCGGATGAGGGTGTTCACCGCCCCGGCCCGGGTGGCCCGCTCGCTCAATTCGTCCACCAGACGATAGGCCTCTTCCTTGAACGGCACGGTGACATTGGCGCCCTTGCCCTGCTCGAAGAAGCGCCGGGCGAACCCGGGGAAATCGTCCAGCGGCGCCAGCTGCGCGTCATAGACCAGCGCCTCGCCGGTCTGCTCGGCGAACAGGCGGTGGATCAGCGGCGACTTGCTGTGGCCGATGGGGTTGCCGAATACGCAATAGCGGTCCATGGGTCGTTCCTTTGCCGGGGTGGGACGGGACGAGCGGACTCGTCCCCTGCGGGCGGGTCAGGGCTGGCCGGCGAGCCAATCGCGGTCGGCGAGGAAATACTCGGTCAGGCGCGCTCCTCNNNNCCGGCC

Allele: 121

>guaA-IISR51NGGNNNNNANACCCCANNGACTNNNNGGCCCTGCTGGTGTGGGTGACTCCGGGGGGGAATTTCAGCCGTGGTAGCCGCGGATCTCTGTCTNCNCGANGAGTGTGATGAGCACGGCGACAAGGTCACCGAGATGCCGGCCGGCTTCCACATCCTGGCCAGCACCCCGAGCTGCCCGATCGCCGCCATGGCCGACGATGCCCGCGCCTACTACGGCGTGCAATTCCACCCGGAAGTCACCCACACCAAGCAGGGCCTGCGCATTCTCTCGCGCTTCGTCCTCGACATCTGCGGTTGCGCCGCGCTGTGGACCCCCTCGAACATCGTCGACGACGCCATCGCCACCGTGCGCGCCCAGGTCGGTTCCTCCAAGGTCCTGCTAGGCCTCTCCGGCGGCGTGGACTCCTCGGTGGTCGCCGCGCTGCTGCACAAGGCCATCGGCGACCAACTGACCTGCGTGTTCGTCGACAACGGCCTGCTGCGCCTGCACGAAGGCGACCAGGTGATGGCCATGTTCGCCGAGAACATGGGCGTGAAGGTGATCCGCGCCAACGCCGAGGACAAGTTCCTCGGCCGCCTGGCCGGCGTCGCCGACCCGGAAGAGAAGCGCAAGATCATCGGCCGCACCTTCATCGAAGTCTTCGACGAAGAAGCCACCAAGCTGCAGGACGTGAAGTTCCTCGCCCAGGGCACCATCTACCCCGACGTGATCGAGTCGGCCGGCGCCAAGACCGGCAAGGCCCACGTGATCAAGTCGCACCACAACGTCGGCGGCCTGCCGGAGGACATGCAGTTCGAACTGGTCGAGCCGCTGCGCGAACTGTTCAAGGACGAAGTGCGCAAGATCGGCCTGGAGCTGGGCCTGCCCTACGACATGGTCTACCGCCACCCGTTCCCCGGCCCGGGCCTGGGCGTGCGCATCCTCGGCGAGGTGAAGAAGGAGTACGCCGACCTGCTGCGCCAGGCCGACCACATCTTCATCGAGGAACTGCGCGCCTTCGACTGGTACCACAGACAGCCCGGGGGCGTTCAGTTCNATGGCTCAAAGATCTAGTAACGGCTTGCAAATTCNGCCCGGGATANTCACNCCANTACCATGCCNNNAGGCGCATTCC

Allele: 36

>mutL-IISR51NCGAGGTCATGGTNNNCGCGCAACCGAANTATCGAGGCAAGCGACTTCGCCNACGAAGCCGAGGCTTCATCACCCGCTCCATGNCNTCAATAGTCCCCCGCGTGAGTGTCGAGCGTCCCGCTCGGTGGCCAAGGAACTCTGGAAAACAGCTTGACGCCGGTTCCCGGCGCATTGATGTGGAGGTCGAGCAGGGCGGCATCAAGTTGCTGCGAGTGCGCGACGACGGTCGCGGCATCCCCGCCGACGACCTGCCGCTGGCCCTGGCTCGCCACGCCACCAGCAAGATCCGCGAGCTGGAAGACCTGGAGCGGGTGATGAGCCTCGGCTTCCGTGGCGAGGCGCTGGCCTCGATCAGTTCGGTAGCGCGCCTGACCATGACCTCGCGTACCGCCGACGCCGGCGAAGCCTGGCAGGTGGAAACCGAGGGCCGTGACATGCAGCCGCGGGTACAGCCGGCGGCGCACCCGGTGGGGACCAGCGACGAGGTTCGCGACCTGTTCTTCAACACCCCGGCGCGGCGCAAGTTCCTGCGCGCCGAGAAGACCGAGTTCGACCATCTGCAGGAAGTCATCAAGCGCCTGGCGCTGGCCCGTTTCGACGTGGCTTTCCACCTGCGCCACAACGGCAAGACCATCTTCGCCCTGCACGAGGCGCGAGACGAGCTGGCCCGCGCGCGCCGGGTCGGCGCGGTGTGCGGCCAGGCATTCCTCGAGCAGGCGCTGCCGATCGAGGTCGAGCGCAACGGCCTGCACCTGTGGGGCTGGGTCGGCTTGCCGACCTTCTCCCGCAGCCAGCCGGACCTGCAGTACTTCTATGTGAACGGGCGCATGGTGCGCGACAAGCTGGTCGCCCACGCGGTGCGCCAGGCTTATCGCGACGTGCTGTACAACGGCCGGCATCCGACCTTCGTGCTGTTCTTCGAAGTCGATCCGGCGGTGGTGGACGTCAACGTGCACCCGACCAAGCACGAAGTTCGCTTCCGTGACAGCCGGATGGTCCATGACTTCCTCTANGGGGNCACCNGAA

Allele: 5

>nuoD-IISR51CCCCCGNTNGCCNACGCCACTGGTACGAGCGCGAAGTGTGGGACATGTACGGCATCACCTTCACCGGCCACCCGCACCTGACCCGCATGCTGATGCCGCCGACCTGGCAGGGCCATCCGCTGCGCAAGGACTACCCGGCGCGCGCCACCGAGTTCGATCCCTACTCCCTGTCCGCCGCCAAGCAGGACCTGGAGCAGGAAGCCCTGCGCTTCAAGCCGGAAGACTGGGGCATGAAGCGCCACGGCGAGAACGAGGACTACATGTTCCTCAACCTCGGCCCGAACCACCCGTCCGCCCACGGCGCGTTCCGCATCATCCTGCAACTGGACGGCGAGGAGATCATCGACTGCGTCCCGGAGATCGGCTACCACCACCGCGGCGCCGAGAAGATGGCCGAGCGCCAGTCCTGGCACAGTTTCATCCCCTACACCGACCGCATCGACTACCTCGGCGGGGTGATGAACAACCTGCCCTACGTACTCTCGGTGGAGAAGCTCGCCGGGATCAAGGTGCCCCAGCGGGTCGACGTGATCCGGATCATGATGGCGGAGTTCTTCCGTATCCTGAACCACCTGCTGTACCTGGGCACCTATATCCAGGACGTCGGCGCCATGACCCCGGTGTTCTTCACCTTCACCGACCGCCAGCGCGCCTACAAGGTGGTCGAGGCCATCACCGGCTTCCGCCTGCACCCGGCCTGGTACCGCATCGGCGGCGTCGCCCACGACCTGCCGCGCGGCTGGGACAAGCTGGTCCGCGAGTTCCTCGACTGGATGCCCAAGCGCCTCGACGAGTACGAGACCGCGGCCCTGAAGAACAGCATCTTGCGCGGCCGGACCATCGGCGTCGCCCAGTACAACACCAAGGAAGCCCTCGAGTGGGGCACCACCGGCGCCGGCCTGCGCGCCACCGGTTGCGACTTCGACCTGCGCAAGGCCCGCCCCTACTCCCGGCTACGAGAACTTCGAGTTCGAGGTACCGCTGGCCCACACGGCGACGCTACGACGCTGCNTNNNNNN

Allele: 4

>ppsA-IISR51NCGNCAAAATGNTTCTTGCNTATNNGNNNNTTTTGNGTGCTTCCTTNNGTATGTANGACGTCGATCGTGCCGATCGCGCCCGCTTCGCCTGAGCGACGCCGAGGTCACCGAGCTGGCCCAAGCAGGCCATGATCATCGAGAAGCACTATGGCCGCCCGATGGACATCGAATGGGCCAAGGACGGTGACGACGGCAAGCTGTACATCGTCCAGGCACGCCCGGAAACCGTGAAGAGCCGCGCCAGCGCCACGGTCATGGAGCGCTACCTGCTGAAAGAGAAGGGGACCGTCCTGGTGGAAGGGCGTGCCATCGGCCAGCGCATCGGTGCCGGTCCGGTCAAGGTGATCAACGACGTGTCGGAAATGGACAAGGTCCAACCGGGTGACGTCCTGGTCTCCGACATGACCGACCCGGACTGGGAGCCAGTGATGAAGCGCGCCAGCGCCATCGTCACCAACCGCGGCGGGCGCACCTGCCACGCGGCGATCATCGCTCGCGAACTGGGCATCCCGGCGGTGGTCGGTTGCGGCAACGCCACCCAGATCCTGCAGGATGGCCAGGGGGTGACCGTTTCCTGTGCCGAAGGCGATACCGGCTTCATCTTCGAAGGCGAACTCGGTCCCGATGTGCGCAAGAACTCGGTCGACGCCATGCCCGACCTTCCGTTCAAGATCATGATGAACGTCGGCAATCCCGATCGCGCTTTCGATTTCGCCCAGTTGCCGAACGAAGGCGTGGGCCTGGCCCGCCTCGAATTCATCATCAACCGCATGATCGGCGTGCATCCCAAGGCGTTGCTGAACTTCGCCGGCCTGCCGGCGGACATCAAGGAAAGCGTGGAGAAGCGCATCGCCGGCTATCCCGATCCGGTCGGCTTCTACGTCGAGAAGCTGGTGGAGGGCATCAGCACCCTGGCCGCGGCGTTCTGGCCGAAGAAGGTCATCGTGCGCCTGTCCGACTTCAAGTCCAATGAGTACGCCACCTGATCGGCGCAAGCTCTACGAGCCGGAGAAAAGAGCAACACCTNCNACCATTGNNNGCANNNCAANN

Allele: 15

P. aeruginosa IISR51

>trpE-IISR51GATACCGCCGCCGGCCTGCACGTGGAGTTCACCGTTCTTGATCACCGCGGTGCGGATGGCAATGGCGGTGTCCATGTTGCCGTTCCATGCCAGGTAGCCGACCGCGCCGCCGTAGACTCCACGCTTGACCGGCTCCAGCTCGTCGATGATCTCCATGGCGCGGATCTTCGGCGCGCCGGACAGCGTACCCGCCGGCAGGATCGCCCGCAGCGCGTCCATCGCGCTGAGCCCCTCGCGCAATTGCCCGGTGACGTTGGACACGATGTGCATGACGTTGGAGTAACGTTCGATCACCATTTTTTCGGTGACCTTCACCGCGCCGATATCGGACACCCGCCCCACGTCGTTGCGCCCCAGGTCGATCAGCATCAGGTGCTCGGCGATCTCCTTGGCGTCCGACAGCAGATCCTGCTCCAGCGCCAGGTCGGCCTCTTCGTTGATCCCGCGCGGACGGGTACCGGCGATCGGGCGCACCGTCACCAGGCCATCCTCGACCCGTACCAGCACCTCCGGCGAGCTGCCCACGACATGGAAGTCGCCGAAGTTGAAGAAGTACATGTAGGGCGTCGGATTGAAACAGCGCAGCGCGCGGTACAGGTCGATGGGCGCCGCCTTGAATTCGATGGACATGCGCTGCGACGGCACCACCTGCATGCAGTCGCCGGCCAGGATGTAGTCCTTGATCCTTCCTACCGCGTTTTCATAGTCCTCGCGGGTG

Allele:8

>acsA-IISR13TGCTATCACTCTNGGGTTTTATNGTCGCTGACCCACGAGCGGGTCTTCGACTACCGTCCGGGCGAAGTCTACTGGTGCACCGCCGACATCGGCTGGGTCACCGGCCACACCTACATCGTCTATGGCCCGCTGGCCAACGGCGCCACCACCATTCTGTTCGAGGGCGTGCCGAACTACCCCGACGTGACCCGCGTGGCGAAAATCATCGACAAGCACAAGGTCAACATCCTCTACACCGCGCCGACCGCGATCCGCGCGATGATGGCCGAAGGCAAGGCGGCGGTGGCCGGTGCCGACGGTTCCAGCCTGCGTCTGCTCGGTTCGGTGGGCGAGCCGATCAACCCGGAAGCCTGGCAGTGGTACTACGAGACCGTCGGCCAGTCGCGCTGCCCGATCGTCGACACCTGGTGGCAGACCGAGACCGGCGCCTGCCTGATGACCCCGCTGCCGGGCGCCCACGCGATGAAGCCGGGCTCTGCAGCCAAGCCGTTCTTCGGCGTGGTACCGGCGCTGGTGGACAACCTCGGCAACCTGATCGAGGGCGCCGCCGAGGGCAACCTGGTGATCCTCGACTCCTGGCCGGGCCAGGCGCGGACCCTGTTCGGCGACCATGACCGCTTCGTCGACACCTACTTCAAGACCTTCAAGGGCATGTACTTCACCGGCGACGGCGCGCGCCGCGACGAGGACGGCTACTACTGGATCACCGGGCGGGTCGACGACGTGCTCAACGTCTCCGGCCACCGCATAAGCACCGCCGAGGTGGAAAGCGCGATGGTCGCCCACCCGAAGGTCGCCGAGGCGGCGGTGGTCGGCATGCAGCACGACATCAAGGGGCACCCATTCTCTTTAGAGNGAGGGGGGCGTC

Allele: 15

>aroE-IISR13GGNTTNNNGGGACGGATTTGAGACGTACGTCTCAAATGCTGCCTGACAACTCAGACAGTTGCCAATCGCCGGCGCAGCGTCTCCAACACTGGCGCCGAGGCAGGACGCACGCCACGCCAGAGGAAGAATGCCTCGGCGGCCTGCTCCACCAGCATGCCCAGGCCATCCAGGGTACGCGCCGCACCGCGTTCGGCGGCCCAGCGGTTGAAGGCGGTCGGTTCCTTGGCATACATCATGTCGTAGCAAACGGTACGGCCGGGCTCGATCACGCTCTGCGCCAGCGGCGGCACGTCGCCGGCAAGACTGGCCGAGGTGCCGTTGACGACCAGGTCGAAAGGCCCTTCGACCTCGGCGAAACCGCAGCCGCGCACCGCGCCGAGATCGGCGAACCGCTCGGCCAGATCCACGGCCTTCCGCGCCGTGCGGTTGGCGATCAGCAGCTCCGCCGGGCACTCGCCGAGGAAGGGTTCGAGCACCCCGCGCACCGCACCGCCGGCGCCGAGCAGGAGAACCCGCTTGCCGCGCAGCTCGACCCCGGCGTTCGCCGTCAGGTCCCGCAGCAAGCCCGCGCCGTCGGTGTTGTCGCCGCGCAGGCGACCGTCGGCCAGGCGGATGAGGGTGTTCAACGCCCCGGCCCGGGTGGCCCGCTCGCTCAATTCGTCCACCAGACGATAGGCCTCTTCCTTGAACGGCACGGTGACATTGGCGCCCTTGCCCTGCTCGAAGAAGCGCCGGGCGAACCCGGGGAAATCGTCCAGCGGCGCCAGCTGCGCGTCATAGACCAGCGCCTCGCCGGTCTGCTCGGCGAACAGGCGGTGGATCAGCGGCGACTTGCTGTGGCCGATGGGGTTGCCGAATACGCAATAGCGGTCCATGGGTCGTTCCTTTGCCGGGGTGGGNAACGGGACGAGCGAACTCGTCCCCTGGCGGGCGGGTCAGGGCTGGCCGGCGAGCCAATCGCGGTCGGCGAGGAAATACTCGGTCAGGCGCGCTCCCCGNNNCGGGCA

Allele: 121

>mutL-IISR13NGNNGCGGTACCAGAGGTCATGTCNGCGCGNACCAACNATCNCGCCAGCGCCTCNCCANNANGCCGAGGCTCATCACCGCTTCTCAGNATCTTNGTCCGCGTGAGCTGTCGAGCGTCCCGCCTCGGTGGCCAAGGAACTCTGGAAAACAGCTTGACGCCGGTTCCCGGCGCATTGATGTGGAGGTCGAGCAGGGCGGCATCAAGTTGCTGCGAGTGCGCGACGACGGTCGCGGCATCCCCGCCGACGACCTGCCGCTGGCCCTGGCTCGCCACGCCACCAGCAAGATCCGCGAGCTGGAAGACCTGGAGCGGGTGATGAGCCTCGGCTTCCGTGGCGAGGCGCTGGCCTCGATCAGTTCGGTAGCGCGCCTGACCATGACCTCGCGTACCGCCGACGCCGGCGAAGCCTGGCAGGTGGAAACCGAGGGCCGTGACATGCAGCCGCGGGTACAGCCGGCGGCGCACCCGGTGGGGACCAGCGTCGAGGTTCGCGACCTGTTCTTCAACACCCCGGCGCGGCGCAAGTTCCTGCGCGCCGAGAAGACCGAGTTCGACCATCTGCAGGAAGTCATCAAGCGCCTGGCGCTGGCCCGTTTCGACGTGGCTTTCCACCTGCGCCACAACGGCAAGACCATCTTCGCCCTGCACGAGGCGCGAGACGAGCTGGCCCGCGCGCGCCGGGTCGGCGCGGTGTGCGGCCAGGCATTCCTCGAGCAGGCGCTGCCGATCGAGGTCGAGCGCAACGGCCTGCACCTGTGGGGCTGGGTCGGCTTGCCGACCTTCTCCCGCAGCCAGCCGGACCTGCAGTACTTCTATGTGAACGGGCGCATGGTGCGCGACAAGCTGGTCGCCCACGCGGTGCGCCAGGCTTATCGCGACGTGCTGTACAACGGCCGGCATCCGACCTTCGTGCTGTTCTTCGAAGTCGATCCGGCGGTGGTGGACGTCAACGTGCACCCGACCAAGCACGAAGTTCGCTTCCGTGACAGCCGGATGGTCCATGACTTCCTCTAAGAGGAGACNCCTNNN

Allele: 5

>mutL-IISR13NGNNGCGGTACCAGAGGTCATGTCNGCGCGNACCAACNATCNCGCCAGCGCCTCNCCANNANGCCGAGGCTCATCACCGCTTCTCAGNATCTTNGTCCGCGTGAGCTGTCGAGCGTCCCGCCTCGGTGGCCAAGGAACTCTGGAAAACAGCTTGACGCCGGTTCCCGGCGCATTGATGTGGAGGTCGAGCAGGGCGGCATCAAGTTGCTGCGAGTGCGCGACGACGGTCGCGGCATCCCCGCCGACGACCTGCCGCTGGCCCTGGCTCGCCACGCCACCAGCAAGATCCGCGAGCTGGAAGACCTGGAGCGGGTGATGAGCCTCGGCTTCCGTGGCGAGGCGCTGGCCTCGATCAGTTCGGTAGCGCGCCTGACCATGACCTCGCGTACCGCCGACGCCGGCGAAGCCTGGCAGGTGGAAACCGAGGGCCGTGACATGCAGCCGCGGGTACAGCCGGCGGCGCACCCGGTGGGGACCAGCGTCGAGGTTCGCGACCTGTTCTTCAACACCCCGGCGCGGCGCAAGTTCCTGCGCGCCGAGAAGACCGAGTTCGACCATCTGCAGGAAGTCATCAAGCGCCTGGCGCTGGCCCGTTTCGACGTGGCTTTCCACCTGCGCCACAACGGCAAGACCATCTTCGCCCTGCACGAGGCGCGAGACGAGCTGGCCCGCGCGCGCCGGGTCGGCGCGGTGTGCGGCCAGGCATTCCTCGAGCAGGCGCTGCCGATCGAGGTCGAGCGCAACGGCCTGCACCTGTGGGGCTGGGTCGGCTTGCCGACCTTCTCCCGCAGCCAGCCGGACCTGCAGTACTTCTATGTGAACGGGCGCATGGTGCGCGACAAGCTGGTCGCCCACGCGGTGCGCCAGGCTTATCGCGACGTGCTGTACAACGGCCGGCATCCGACCTTCGTGCTGTTCTTCGAAGTCGATCCGGCGGTGGTGGACGTCAACGTGCACCCGACCAAGCACGAAGTTCGCTTCCGTGACAGCCGGATGGTCCATGACTTCCTCTAAGAGGAGACNCCTNNN

Allele: 5

>nuoD-IISR13NANTGCCCACGCACTGGTACGAGCGCGAAGTGTGGGACATGTACGGCATCACCTTCACCGGCCACCCGCACCTGACCCGCATGCTGATGCCGCCGACCTGGCAGGGCCATCCGCTGCGCAAGGACTACCCGGCGCGCGCCACCGAGTTCGATCCCTACTCCCTGTCCGCCGCCAAGCAGGACCTGGAGCAGGAAGCCCTGCGCTTCAAGCCGGAAGACTGGGGCATGAAGCGCCACGGCGAGAACGAGGACTACATGTTCCTCAACCTCGGCCCGAACCACCCGTCCGCCCACGGCGCGTTCCGCATCATCCTGCAACTGGACGGCGAGGAGATCATCGACTGCGTCCCGGAGATCGGCTACCACCACCGCGGCGCCGAGAAGATGGCCGAGCGCCAGTCCTGGCACAGTTTCATCCCCTACACCGACCGCATCGACTACCTCGGCGGGGTGATGAACAACCTGCCCTACGTACTCTCGGTGGAGAAGCTCGCCGGGATCAAGGTGCCCCAGCGGGTCGACGTGATCCGGATCATGATGGCGGAGTTCTTCCGTATCCTGAACCACCTGCTGTACCTGGGCACCTATATCCAGGACGTCGGCGCCATGACCCCGGTGTTCTTCACCTTCACCGACCGCCAGCGCGCCTACAAGGTGGTCGAGGCCATCACCGGCTTCCGCCTGCACCCGGCCTGGTACCGCATCGGCGGCGTCGCCCACGACCTGCCGCGCGGCTGGGACAAGCTGGTCCGCGAGTTCCTCGACTGGATGCCCAAGCGCCTCGACGAGTACGAGACCGCGGCCCTGAAGAACAGCATCTTGCGCGGCCGGACCATCGGCGTCGCCCAGTACAACACCAAGGAAGCCCTCGAGTGGGGCACCACCGGCGCCGGCCTGCGCGCCACCGGTTGCGACTTCGACCTGCGCAAGGCCCGCCCCTACTCCGGCTACGAGAACTTCGAGTTCGAGGTACCGCTGGCCCACACGGCGACGCTACGACGCTGCANNNNGAAN

Allele: 4

>ppsA-IISR13NTCGAGGGCCATATNCNNTCGATGATCTTGCCTNCTNNGGTCGCNNNNTNNCAGTAGTGACGTCGATCGTGCCGATCGCGCCGCTTCGCCCTGAGCGACGCCGAGGTCACCGAGCTGGCCAAGCAGGCCATGATCATCGAGAAGCACTATGGCCGCCCGATGGACATCGAATGGGCCAAGGACGGTGACGACGGCAAGCTGTACATCGTCCAGGCACGCCCGGAAACCGTGAAGAGCCGCGCCAGCGCCACGGTCATGGAGCGCTACCTGCTGAAAGAGAAGGGGACCGTCCTGGTGGAAGGGCGTGCCATCGGCCAGCGCATCGGTGCCGGTCCGGTCAAGGTGATCAACGACGTGTCGGAAATGGACAAGGTCCAACCGGGTGACGTCCTGGTCTCCGACATGACCGACCCGGACTGGGAGCCAGTGATGAAGCGCGCCAGCGCCATCGTCACCAACCGCGGCGGGCGCACCTGCCACGCGGCGATCATCGCTCGCGAACTGGGCATCCCGGCGGTGGTCGGTTGCGGCAACGCCACCCAGATCCTGCAGGATGGCCAGGGGGTGACCGTTTCCTGTGCCGAAGGCGATACCGGCTTCATCTTCGAAGGCGAACTCGGTTTCGATGTGCGCAAGAACTCGGTCGACGCCATGCCCGACCTTCCGTTCAAGATCATGATGAACGTCGGCAATCCCGATCGCGCTTTCGATTTCGCCCAGTTGCCGAACGAAGGCGTGGGCCTGGCCCGCCTCGAATTCATCATCAACCGCATGATCGGCGTGCATCCCAAGGCGTTGCTGAACTTCGCCGGCCTGCCGGCGGACATCAAGGAAAGCGTGGAGAAGCGCATCGCCGGCTATCCCGATCCGGTCGGCTTCTACGTCGAGAAGCTGGTGGAGGGCATCAGCACCCTGGCCGCGGCGTTCTGGCCGAAGAAGGTCATCGTGCGCCTGTCCGACTTCAAGTCCAATGAGTACGCCAACCTGATCGGCGCAAGCTCTACGAGCCTGAGNNNNTACCTNNACCATNNCGCAGCNACCANNANNNNTTNGCAGAAAGG

ppsA: 15

>trpE-IISR13GGGGGGNGGNTNTCGCTTACCGCGAGGACTATGAAACGCGGTAGGAAGGATCAAGGACTACATCCTGGCCGGCGACTGCATGCAGGTGGTGCCGTCGCAGCGCATGTCCATCGAATTCAAGGCGGCGCCCATCGACCTGTACCGCGCGCTGCGCTGTTTCAATCCGACGCCCTACATGTACTTCTTCAACTTCGGCGACTTCCATGTCGTGGGCAGCTCGCCGGAGGTGCTGGTACGGGTCGAGGATGGCCTGGTGACGGTGCGCCCGATCGCCGGTACCCGTCCGCGCGGGATCAACGAAGAGGCCGACCTGGCGCTGGAGCAGGATCTGCTGTCGGACGCCAAGGAGATCGCCGAGCACCTGATGCTGATCGACCTGGGGCGCAACGACGTGGGGCGGGTGTCCGATATCGGCGCGGTGAAGGTCACCGAAAAAATGGTGATCGAACGTTACTCCAACGTCATGCACATCGTGTCCAACGTCACCGGGCAATTGCGCGAGGGGCTCAGCGCGATGGACGCGCTGCGGGCGATCCTGCCGGCGGGTACGCTGTCCGGCGCGCCGAAGATCCGCGCCATGGAGATCATCGACGAGCTGGAGCCGGTCAAGCGTGGAGTCTACGGCGGCGCGGTCGGCTACCTGGCATGGAACGGCAACATGGACACCGCCATTGCCATCCGCACCGCGGTGATCAAGAACGGTGAACTCCACGTGCAGGCCGGCGGCGGTATCGTTGCCGACTCGGTGCCGGCGCTGGAGTGGGAAGAACCATCAACAGGCGCCGGACCGAGTCGGCACGATCCGCCACAGCTGCAGTGGAGTTCACGGTTTGATCACGGCGTGCGAGGCATGCGTTCATGTGGCTTTCAGCGGTAGCACGCGCCNAAANNCGCTGGACGGCGCGCGNCNTAATTCNGGCGGNAAATTACGCGCCGNNNNGNC

Allele: 8

P. aeruginosa IISR13

>acsA-IISR853TATTCTGGTGTTTCGCCTCGCTGACCCACGAGCGGGTCTTCGACTACCGTCCGGGCGAAGTCTACTGGTGCACCGCCGACATCGGCTGGGTCACCGGCCACACCTACATCGTCTATGGCCCGCTGGCCAACGGCGCCACCACCATTCTGTTCGAGGGCGTGCCGAACTACCCCGACGTGACCCGCGTGGCGAAAATCATCGACAAGCACAAGGTCAACATCCTCTACACCGCGCCGACCGCGATCCGCGCGATGATGGCCGAAGGCAAGGCGGCGGTGGCCGGTGCCGACGGTTCCAGCCTGCGTCTGCTCGGTTCGGTGGGCGAGCCGATCAACCCGGAAGCCTGGCAGTGGTACTACGAGACCGTCGGCCAGTCGCGCTGCCCGATCGTCGACACCTGGTGGCAGACCGAGACCGGCGCCTGCCTGATGACCCCGCTGCCGGGCGCCCACGCGATGAAGCCGGGCTCCGCGGCCAAGCCGTTCTTCGGCGTGGTACCGGCACTGGTGGACAATCTCGGCAACCTGATCGAGGGCGCCGCCGAGGGCAACCTGGTGATCCTCGACTCCTGGCCGGGCCAGGCGCGGACCCTGTTCGGCGACCATGACCGCTTCGTCGACACCTACTTCAAGACCTTCAAGGGCATGTACTTCACCGGCGACGGCGCGCGCCGCGACGAGGACGGCTACTACTGGATCACCGGGCGGGTCGACGACGTGCTCAACGTCTCCGGCCACCGCATGGGCACCGCCGAGGTGGAAAGCGCGATGGTCCGCCCACCCCGAAGGTCGCCGAGGCGGCGGTGGTCGGCATGCAGCACGACATCAAGGAGGGGATTTTTTTTTTNATAAGCCNCCCTGTTAGCCNGGTAACTTNGGTGGGGCACATGGTTCACTNGTCGTCCAGCCGTGGCGGAGACTTCAGAGTCTCACCGCGTGATCTTTNTAGCCNCNCTNCGCGNCNCCGNTCAATCANGTCNNGANGTTTGANNNAGTTTCGACAAAACGCTTAGNATGCCTACAGGGACCNNN

Allele: 22

>aroE-IISR853GGAGAGGGTAAANCCATTTTGAGACGTACGTCTCAAATGCCGCCTGACAACTCAGACAGTTGCCAACTGTCGGCGCAGCGTCTCCAACACTGGCGCCGAGGCAGGACGCACGCCGCGCCAGAGGAAGAATGCCTCGGCGGCCTGCTCCACCAGCATGCCCAGGCCATCCAGGGTACGCGCCGCACCGCGTTCGGCGGCCCAGCGGTTGAAGGCAGTCGGTTCCTTGGCATACATCATGTCGTAGCAGACGGTACGGCCGGGCTCGATCACGCTCTGCGCCAGCGGCGGCACGTCGCCGGCAAGACTGGCCGAGGTGCCGTTGACGATCAGGTCGAAAGGCCCTTCGACCTCGGCGAAACCGCAGCCGTGCACCGCGCCGAGGTCGGCGAACCGCTCGGCCAGGTCCACGGCCTTCCGCGCCGTGCGGTTGGCGATCAGCAACTCCGCCGGGCACTCGCCGAGGAAGGGTTCGAGCACCCCACGCACCGCACCGCCGGCGCCGAGCAGGAGAACCCGCTTGCCGCGCAGCTCGACCCCGGCGTTCGCCGTCAGGTCCCGCAGCAGGCCGGCGCCGTCGGTGTTGTCGCCGCGCAGGCGACCGTCGGCGAGGCGGATCAGGGTGTTCACCGCCCCGGCCCGGGTGGCCCGCTCGCTCAACTCGTCCACCAGACGATAGGCCTCTTCCTTGAACGGCACGGTGACATTGGCGCCCTTGCCCTGCTCGAAGAAGCGCCGGGCGAACCCGGGGAAATCGTCCAGCGGCGCCAGTTGCGCGTCATAGACCAGCGCCTCGCCGGTCTGCTCGGCGAACAGGCGGTGGATCAGCGGCGACTTGCTGTGGCCGATGGGGTTGCCGAATACGCAATAGCGGTCCATGGGTCGTTCCTTTGCCGGAGTGGGGACGGGACGAGCGGACTCGTCCCCTGCGGGCGGGTCAGGGCTGGCCGGCGAGCCAATCGCGGTCGGCGAGGAAATACTCGGTCAGGCGCGCTCCCCNCCCNCGGNN

Allele: 5

>guaA-IISR853NGTAGTGAGNAAACCAAAGAACTGGCCNNCCTGCTGGTGNTGGGTGACTTCCGGGGGAATTCCAGCGGTCGTGGGCGCGGGATTTCTGGCCTTNNAACGTGTGATGAGCCACGGCGACAAGGTCACCGAGATGCCGGCCGGCTTCCACATCCTGGCCAGCACCCCGAGCTGCCCGATCGCCGCCATGGCCGACGATGCCCGCGCCTACTACGGCGTGCAATTCCACCCGGAAGTCACCCACACCAAGCAGGGCCTGCGCATTCTCTCGCGCTTCGTCCTCGACATCTGCGGTTGCGCCGCGCTGTGGACCCCCTCGAACATCGTCGACGACGCCATCGCCACCGTGCGCGCCCAGGTCGGTTCCTCCAAGGTCCTGCTAGGCCTCTCCGGCGGCGTGGACTCCTCGGTGGTCGCCGCGCTGCTGCACAAGGCCATCGGCGACCAACTGACCTGCGTGTTCGTCGACAACGGCCTGCTGCGCCTGCACGAAGGCGACCAGGTGATGGCCATGTTCGCCGAGAACATGGGCGTGAAGGTGATCCGCGCCAACGCCGAGGACAAGTTCCTCGGCCGCCTGGCCGGCGTCGCCGACCCGGAAGAGAAGCGCAAGATCATCGGCCGCACCTTAATCGAAGTTTTCGACGAAGAAGCCACCAAGCTGCAGGACGTGAAGTTCCTCGCCCAGGGCACCATCTACCCCGACGTGATCGAGTCAGCCGGCGCCAAGACCGGCAAGGCCCACGTGATCAAGTCGCACCACAACGTCGGCGGCCTGCCGGAGGACATGCAGTTCGAACTGGTCGAGCCGCTGCGCGAACTGTTCAAGGACGAAGTGCGCAAGATCGGCCTGGAGCTGGGCCTGCCCTACGACATGGTCTACCGCCACCCGTTCCCCGGCCCGGGCCTGGGCGTGCGCATCCTCGGCGAGGTGAAGAAGGAGTACGCCGACCTGCTGCGCCAGGCCGACCACATCTTCATCGAGGAACTGCGCGCCTTCGACTGGTACACAGACAGCCCGGGGCGTTCAGTACCAATGCCAAGNTCTAGCTACGCCNTGCAATTCCGCNNCGTAGTCNCTCCTACACCNGCNNGNNTGCCCANTCCTTGNNNCT

Allele: 83

>mutL-IISRT853AAGTCATGGTCAGGCGGCGCAACCNGGTTATCGAGNCAAGCCGCCTCGCCACGGAAGCGGGGGCTCATCACCCGCTCCTCGTANCATCGTCCGCCGGTGAGGTGGTCGAGCGTCCCGCCTCGGTGGCCAAGGAACTCTGGAAAACAGCCTTGACGCCGGTTCCCGGCGCATTGATGTGGAGGTCGAGCAGGGCGGCATCAAGTTGCTGCGAGTGCGCGACGACGGTCGCGGCATCCCCGCCGACGACCTGCCGCTGGCCCTGGCTCGCCACGCCACCAGCAAGATCCGCGAGCTGGAAGACCTGGAGCGGGTGATGAGCCTCGGCTTCCGTGGCGAGGCGCTGGCCTCGATCAGCTCGGTAGCGCGCCTGACCATGACCTCGCGTACCGCCGACGCCGGCGAAGCCTGGCAGGTGGAAACCGAGGGCCGCGACATGCAGCCGCGGGTACAGCCGGCGGCGCATCCGGTGGGGACCAGCGACGAGGTTCGCGACCTGTTCTTCAACACCCCGGCGCGGCGCAAGTTCCTGCGTGCCGAGAAGACCGAGTTCGACCATCTGCAGGAGGTCATCAAGCGCCTGGCGCTGGCCCGCTTCGACGTGGCTTTCCACCTGCGCCACAACGGCAAGACCATCTTCGCCCTGCACGAGGCGCGAGACGAGCTGGCCCGCGCGCGCCGGGTCGGCGCGGTGTGCGGCCAGGCATTCCTCGAGCAGGCGCTGCCGATCGAGGTCGAGCGCAACGGTCTGCACCTGTGGGGCTGGGTCGGCTTGCCGACCTTCTCCCGCAGCCAGCCGGACCTGCAGTACTTCTATGTGAACGGGCGCATGGTGCGCGACAAGCTGGTCGCCCACGCGGTGCGCCAGGCTTATCGCGACGTGCTGTACAACGGCCGGCATCCGACCTTCGTGCTGTTCTTCGAAGTCGATCCGGCGGTGGTGGACGTCAACGTGCACCCGACCAAGCACGAAGTTCGCTTCCGTGACAGCCGGATGGTCCATGACTTCCTCTATGGGAACCCTGA

Allele: 2

>nuoD-IISR853TNNAGCAGCGGTCGTAGGCGTCGCCGTTGTGGGCCAGCGGTACCTCGAACTCGAAGTTCTCGTAGCCGGAGTAGGGGCGGGCCTTGCGCAGGTCGAAGTCGCAACCGGTGGCGCGCAGGCCGGCGCCGGTGGTGCCCCACTCGAGGGCTTCCTTGGTGTTGTACTGGGCGACGCCGATGGTCCGGCCGCGCAGGATGCTGTTCTTCAGGGCCGCGGTCTCGTACTCGTCGAGGCGCTTGGGCATCCAGTCGAGGAACTCGCGGACCAGCTTGTCCCAGCCGCGCGGCAGGTCGTGGGCGACGCCGCCGATGCGGTACCAGGCCGGGTGCAGGCGGAAGCCGGTGATGGCCTCGACCACCTTGTAGGCGCGCTGGCGGTCGGGGAAGGTGAAGAACACCGGGGTCATGGCGCCGACGTCCTGGATATAGGTGCCCAGGTACAGCAGGTGGTTCAGGATACGGAAGAACTCCGCCATCATGATCCGGATCACGTCGACCCGCTGGGGCACCTTGATCCCGGCGAGCTTCTCCACCGAGAGTACGTAGGGCAGGTTGTTCATCACCCCGCCGAGGTAGTCGATGCGGTCGGTGTAGGGGATGAAACTGTGCCAGGACTGGCGCTCGGCCATCTTCTCGGCGCCGCGGTGGTGGTAGCCGATCTCCGGGACGCAGTCGATGATCTCCTCGCCGTCCAGTTGCAGGATGATGCGGAACGCGCCGTGGGCGGACGGGTGGTTCGGGCCGAGGTTGAGGAACATGTAGTCCTCGTTCTCGCCGTGGCGCTTCATGCCCCAGTCTTCCGGCTTGAAGCGCAGGGCTTCCTGCTCCAGGTCCTGCTTGGCGGCGGACAGGGAGTAGGGATCGAACTCGGTGGCGCGCGCCGGGTAGTCCTTGCGCAGCGGATGGCCCTGCCAGGTCGGCGGCATCAGCATGCGGGTCAGGTGCGGGTGGCCGGTGAAGGTGATGCCGTACATGTCCCACACTTCGCGCTCGTACAGTGGCGTGNCNNNNGGG

Allele: 4

>ppsA-IISR853CTATCGGGGCANTAGNNNNTTGGGCNGTTTTTNNNNGNCNNGTTGNTCTNCGCTNNAGGTAGTGACGTCGATCGCGCCGATCGCGCCCGCTTCGCCTGAGCGACGCCGAGGTCACCGAGCTGGCCAAGCAGGCCATGATCATCGAGAAGCACTATGGCCGCCCGATGGACATCGAATGGGCCAAGGACGGTGACGACGGCAAGCTGTACATCGTCCAGGCACGCCCGGAAACCGTGAAGAGCCGCGCCAGCGCCACGGTCATGGAGCGCTACCTGCTGAAAGAGAAGGGGACCGTCCTGGTGGAAGGGCGTGCCATCGGCCAGCGCATCGGTGCCGGTCCGGTCAAAGTGATCAACGACGTGTCGGAAATGGACAAGGTCCAACCGGGTGACGTCCTGGTCTCCGACATGACCGACCCGGACTGGGAGCCAGTGATGAAGCGCGCCAGCGCCATCGTCACCAACCGCGGCGGGCGCACCTGCCACGCGGCGATCATCGCTCGCGAACTGGGCATCCCGGCGGTGGTCGGTTGCGGCAACGCCACCCAGATCCTGCAGGATGGCCAGGGGGTGACCGTTTCCTGTGCCGAAGGCGATACCGGCTTCATCTTCGAAGGCGAACTCGGTTTCGATGTGCGCAAGAACTCGGTCGACGCCATGCCCGACCTTCCGTTCAAGATCATGATGAACGTCGGCAATCCCGATCGCGCTTTCGATTTCGCCCAGTTGCCGAACGAAGGCGTGGGCCTGGCCCGCCTCGAATTCATCATCAACCGCAAGATCGGCGTGCATCCCAAGGCGTTGCTGAACTTCGCCGGCCTGCCGGCGGACATCAAGGAAAGCGTGGAGAAGCGCATCGCCGGCTATCCCGATCCGGTCGGCTTCTACGTCGAGAAGCTGGTGGAGGGCATCAGCACCCTGGCCGCGGCGTTCTGGCCGAAGAAGGTCATCGTGCGCCTGTCCGACTTCAAGTCCAATGAGTACGCCAACCTGATCGGCGCAGCTCTACACNNGNNGCCCNNACCNGCCCCNCTACTNCACNCAA

Allele: 13

>trpE-IISR853CTCACTTGTCGATATGTCCATGGCGGGATTTCGCGCGCGGAAGAGTGCCCGCTGGAAGATCGCCCGCAGCGGCGTCCACCGCGTTGAGCCCCCCGCGCAATTGCCCGGTGGACGGTCGGGCCCGATGTGCACGGCGTTGGGAGGTAACCTTCCATCANCCATTTTTTTCGTGTGACCTTTCACCGCGCCGATATCGGACACCCGCCCCACGTTCGCGGGGGCCCCCAAGGGCGTGAGCGGCGTTTCGTGCCAGCTTCACCCGCGAGGACTATGAAAACGCGGTAGGAAGGATCAAGGACTACATCCTGGCCGGCGACTGCATGCAGGTGGTGCCGTCGCAGCGCATGTCCATCGAATTCAAGGCGGCGCCCATCGACCTGTACCGCGCGCTGCGCTGTTTCAATCCGACGCCCTACATGTACTTCTTCAACTTCGGCGACTTCCATGTCGTGGGCAGCTCGCCGGAGGTGCTGGTACGGGTCGAGGATGGCCTGGTGACGGTGCGCCCGATCGCCGGTACCCGTCCGCGCGGGATCAACGAAGAGGCCGACCTGGCACTGGAGCAGGATCTGCTGTCGGACGCCAAGGAGATCGCCGAGCACCTGATGCTGATCGACCTGGGGCGCAACGACGTGGGGCGGGTGTCCGATATCGGCGCGGTGAAGGTCACCGAAAAAATGGTGATCGAACGTTACTCCAACGTCATGCACATCGTGTCCAACGTCACCGGGCAATTGCGCGAGGGGCTCAGCGCGATGGACGCGCTGCGGGCGATTCTGCCGGCGGGCACTCTATCCGGCGCGCCGAAGATCCGCGCCATGGAGATCATCGACGAGCTGGAGCCGGTCAAGCGTGGAGTCTACGGCGGCGCGGTCGGCTACCTGGCATGGAACGGCAACATGGACACCGCCATTGCCATCCGCACCGCGGTGATCAAGAACGGTGAACTCCACGTGCAGGCCGGCGGCGGTATCGTTGCCGACTCGGTGCCCGCGCTGGAGTGGGAAGAAACCATCACACCCCCCGGGACCAAATCGGCACGATTCCGCCTCGCTGCAGTGAGTTAACCGTTTGGATGACGCGTTCGGAAGGCATGGCGGTTTCATGTGCNTTCAGCNGGTNCNNCGNNNCTAAACCCGCTGGACGGNNNNNTCATNTATTCGGGGCGAANTNGGCGCNAAGAGGGCNGANAAATGCCGCNGCTTGCCNTGNNGCTCANNTGNACGAATTNNNACAAATTGANN

Allele: 7

P. aeruginosa IISR853

>acsA-GEB9NTTGTNCAAAAGAATTGCTGCCCCTTGATGGTCGTGCTGCATGCCGACCACCGCCGCCTCGGCGACCTTCGGGTGGGCGACCATCGCGCTTTCCACCTCGGCGGTGCCCATGCGGTGGCCGGAGACGTTGAGCACGTCGTCGACCCGCCCGGTGATCCAGTAGTAGCCGTCCTCGTCGCGGCGCGCGCCGTCGCCGGTGAAGTACATGCCCTTGAAGGTCTTGAAGTAGGTGTCGACGAAGCGGTCATGGTCGCCGAACAGGGTCCGCGCCTGGCCCGGCCAGGAGTCGAGGATCACCAGGTTGCCCTCGGCGGCGCCCTCGATCAGGTTGCCGAGATTGTCCACCAGTGCCGGTACCACGCCGAAGAACGGCTTGGCCGCGGAGCCCGGCTTCATCGCGTGGGCGCCCGGCAGCGGGGTCATCAGGCAGGCGCCGGTCTCGGTCTGCCACCAGGTGTCGACGATCGGGCAGCGCGACTGGCCGACGGTCTCGTAGTACCACTGCCAGGCTTCCGGGTTGATCGGCTCGCCCACCGAACCGAGCAGACGCAGGCTGGAACCGTCGGCACCGGCCACCGCCGCCTTGCCTTCGGCCATCATCGCGCGGATCGCGGTCGGCGCGGTGTAGAGGATGTTGACCTTGTGCTTGTCGATGATTTTCGCCACGCGGGTCACGTCGGGGTAGTTCGGCACGCCCTCGAACAGAATGGTGGTGGCGCCGTTGGCCAGCGGGCCATAGACGATGTAGGTGTGGCCGGTGACCCAGCCGATGTCGGCGGTGCACCAGTAGACTTCGCCCGGACGGTAGTCGAAGACCCGCTCGTGGGTCAGCGAGGTAANANCCNGAGATCGA

Allele: 22

>aroE-GEB9AGCCGGGCAGAGAGGAAGGCGCCTGACCGAGTATTTCCTCGCCGACCGCGATTGACTCGCCGGCCAGCCCTGACCAGCCCGCAGGGGACGAGTCCGCTCGTCCCGTCCCACTCCGGCAAAGGAACGACCCATGGACCGCTATTGCGTATTCGGCAACCCCATCGGCCACAGCAAGTCGCCGCTGATCCACCGCCTGTTCGCCGAGCAGACCGGCGAGGCGCTGGTCTATGACGCGCAACTGGCGCCGCTGGACGATTTCCCCGGGTTCGCCCGGCGCTTCTTCGAGCAGGGCAAGGGCGCCAATGTCACCGTGCCGTTCAAGGAAGAGGCCTATCGTCTGGTGGACGAGTTGAGCGAGCGGGCCACCCGGGCCGGGGCGGTGAACACCCTGATCCGCCTCGCCGACGGTCGCCTGCGCGGCGACAACACCGACGGCGCCGGCCTGCTGCGGGACCTGACGGCGAACGCCGGGGTCGAGCTGCGCGGCAAGCGGGTTCTCCTGCTCGGCGCCGGCGGTGCGGTGCGTGGGGTGCTCGAACCCTTCCTCGGCGAGTGCCCGGCGGAGTTGCTGATCGCCAACCGCACGGCGCGGAAGGCCGTGGACCTGGCCGAGCGGTTCGCCGACCTCGGCGCGGTGCACGGCTGCGGTTTCGCCGAGGTCGAAGGGCCTTTCGACCTGATCGTCAACGGCACCTCGGCCAGTCTTGCCGGCGACGTGCCGCCGCTGGCGCAGAGCGTGATCGAGCCCGGCCGTACCGTCTGCTACGACATGATGTATGCCAAGGAACCGACTGCCTTCAACCGCTGGGCCGCCGAACGCGGTGCGGCGCGTACCCTGGATGGCCTGGGCATGCTGGTGGAGCAGGCCGCCGAGGCATTCTTCCTCTGGCGCGGCGTGCGTCCTGCCTCGGCGCCAGTGTTGGAGACGCTGCGCCGACAGTTGGCAACTGTCTGAGTTGTCAGGCGGCATTTGAGACGTANGTNNCAATTTGGNTTCCATCTG

Allele: 5

>guaA-GEB9GNGAGCGCGAGAGAATNGCGCNNNCCTTCCTTGGTNGGGNTNNATCTCGGGGGGGGAATTGCNNGCCGTCGTAGGCGCGGGATCTGTCGGTCTCTTAAGCGGTGTGATGAGCCACGGCGACAAGGTCACCGAGATGCCGGCCGCGCTTCCACAGTCCGTGGCCAGCAGCCCCGAGCTGCCCGATCGCCGCCATGGCCGACGATGCCCGCGCCTACTACGGCGTGCAATTCCACCCGGAAGTCACCCACACCAAGCAGGGCCTGCGCATTCTCTCGCGCTTCGTCCTCGACATCTGCGGTTGCGCCGCGCTGTGGACCCCCTCGAACATCGTCGACGACGCCATCGCCACCGTGCGCGCCCAGGTCGGTTCCTCCAAGGTCCTGCTAGGCCTCTCCGGCGGCGTGGACTCCTCGGTGGTCGCCGCGCTGCTGCACAAGGCCATCGGCGACCAACTGACCTGCGTGTTCGTCGACAACGGCCTGCTGCGCCTGCACGAAGGCGACCAGGTGATGGCCATGTTCGCCGAGAACATGGGCGTGAAGGTGATCCGCGCCAACGCCGAGGACAAGTTCCTCGGCCGCCTGGCCGGCGTCGCCGACCCGGAAGAGAAGCGCAAGATCATCGGCCGCACCTTCATCGAAGTTTTCGACGAAGAAGCCACCAAGCTGCAGGACGTGAAGTTCCTCGCCCAGGGCACCATCTACCCCGACGTGATCGAGTCAGCCGGCGCCAAGACCGGCAAGGCCCACGTGATCAAGTCGCACCACAACGTCGGCGGCCTGCCGGAGGACATGCAGTTCGAACTGGTCGAGCCGCTGCGCGAACTGTTCAAGGACGAAGTGCGCAAGATCGGCCTGGAGCTGGGCCTGCCCTACGACATGGTCTACCGCCACCCGTTCCCCGGCCCGGGCCTGGGCGTGCGCATCCTCGGCGAGGTGAAGAAGGAGTACGCCGACCTGCTGCGCCAGGCCGACCACATCTTCATCGAGGAACTGCGCGCCTTCGACTGGTACCACAAGACACCCGGGGCGTATCAGCTCCNCGATCACAACTTGTNCGGCTGGCAATTCACGCCGGAATTTCCTCATCAACTCGGGGAAGANCCNNNCCTGGTCCN

Allele: 83

>mutL-GEB9TCAGGTGNTCCCGTAGAGGGAAGGTCATGGACCATCCGGCTGTCACGGAAGCGAACTTCGTGCCTGGTCGGGTGCACGTTGACGTCCACCACCGCCGGATCGACTTCGAAGAACAGCACGAAGGTCGGATGCCGGCCGTTGTACAGCACGTCGCGATAAGCCTGGCGCACCGCGTGGGCGACCAGCTTGTCGCGCACCATGCGCCCGTTCACATAGAAGTACTGCAGGTCCGGCTGGCTGCGGGAGAAGGTCGGCAAGCCGACCCAGCCCCACAGGTGCAGACCGTTGCGCTCGACCTCGATCGGCAGCGCCTGCTCGAGGAATGCCTGGCCGCACACCGCGCCGACCCGGCGCGCGCGGGCCAGCTCGTCTCGCGCCTCGTGCAGGGCGAAGATGGTCTTGCCGTTGTGGCGCAGGTGGAAAGCCACGTCGAAGCGGGCCAGCGCCAGGCGCTTGATGACCTCCTGCAGATGGTCGAACTCGGTCTTCTCGGCACGCAGGAACTTGCGCCGCGCCGGGGTGTTGAAGAACAGGTCGCGAACCTCGACGCTGGTCCCCACCGGATGCGCCGCCGGCTGTACCCGCGGCTGCATGTCGCGGCCCTCGGTTTCCACCTGCCAGGCTTCGCCGGCGTCGGCGGTACGCGAGGTCATGGTCAGGCGCGCTACCGAGCTGATCGAGGCCAGCGCCTCGCCACGGAAGCCGAGGCTCATCACCCGCTCCAGGTCTTCCAGCTCGCGGATCTTGCTGGTGGCGTGGCGAGCCAGGGCCAGCGGCAGGTCGTCGGCGGGGATGCCGCGACCGTCGTCGCGCACTCGCAGCAACTTGATGCCGCCCTGCTCGACCTCCACATCAATGCGCCGGGAACCGGCGTCAAGGCTGTTTTTCCAGGAGTTCCTTGGCCACCGAGGCGGGACGCTCGACCACCTCACCGGCGGGGACATCATGAGATTGGAGCGGGTGATGANCCTTGGCTTCCNGGGAGAGGCCGCTCGCCTCCGATCATCTNTTTGTNCGTCTGACCATGACCTCTCGCN

Allele: 2

>nuoD-GEB9TCTGACATGCAGCGGTCGTAGGCGTCGCCGTGTGGGCCAGCGGTACCTCGAACTCGAAGTTCTCGTAGCCGGAGTAGGGGCGGGCCTTGCGCAGGTCGAAGTCGCAACCGGTGGCGCGCAGGCCGGCGCCGGTGGTGCCCCACTCGAGGGCTTCCTTGGTGTTGTACTGGGCGACGCCGATGGTCCGGCCGCGCAGGATGCTGTTCTTCAGGGCCGCGGTCTCGTACTCGTCGAGGCGCTTGGGCATCCAGTCGAGGAACTCGCGGACCAGCTTGTCCCAGCCGCGCGGCAGGTCGTGGGCGACGCCGCCGATGCGGTACCAGGCCGGGTGCAGGCGGAAGCCGGTGATGGCCTCGACCACCTTGTAGGCGCGCTGGCGGTCGGTGAAGGTGAAGAACACCGGGGTCATGGCGCCGACGTCCTGGATATAGGTGCCCAGGTACAGCAGGTGGTTCAGGATACGGAAGAACTCCGCCATCATGATCCGGATCACGTCGACCCGCTGGGGCACCTTGATCCCGGCGAGCTTCTCCACCGAGAGTACGTAGGGCAGGTTGTTCATCACCCCGCCGAGGTAGTCGATGCGGTCGGTGTAGGGGATGAAACTGTGCCAGGACTGGCGCTCGGCCATCTTCTCGGCGCCGCGGTGGTGGTAGCCGATCTCCGGGACGCAGTCGATGATCTCCTCGCCGTCCAGTTGCAGGATGATGCGGAACGCGCCGTGGGCGGACGGGTGGTTCGGGCCGAGGTTGAGGAACATGTAGTCCTCGTTCTCGCCGTGGCGCTTCATGCCCCAGTCTTCCGGCTTGAAGCGCAGGGCTTCCTGCTCCAGGTCCTGCTTGGCGGCGGACAGGGAGTAGGGATCGAACTCGGTGGCGCGCGCCGGGTAGTCCTTGCGCAGCGGATGGCCCTGCCAGGTCGGCGGCATCAGCATGCGGGTCAGGTGCGGGTGGCCGGTGAAGGTGATGCCGTACATGTCCCACACTTCGNNCTCGTACCAGTCGCGTNGGNCAGNN

Allele: 4

>ppsA-GEB9GGACAGGCGCACGATGACCTTCTTCGGCCAGAACGCCGCGGCCAGGGTGCTGATGCCCTCCACCAGCTTCTCGACGTAGAAGCCGACCGGATCGGGATAGCCGGCGATGCGCTTCTCCACGCTTTCCTTGATGTCCGCCGGCAGGCCGGCGAAGTTCAGCAACGCCTTGGGATGCACGCCGATCATGCGGTTGATGATGAATTCGAGGCGGGCCAGGCCCACGCCTTCGTTCGGCAACTGGGCGAAATCGAAAGCGCGATCGGGATTGCCGACGTTCATCATGATCTTGAACGGAAGGTCGGGCATGGCGTCGACCGAGTTCTTGCGCACATCGAAACCGAGTTCGCCTTCGAAGATGAAGCCGGTATCGCCTTCGGCACAGGAAACGGTCACCCCCTGGCCATCCTGCAGGATCTGGGTGGCGTTGCCGCAACCGACCACCGCCGGGATGCCCAGTTCGCGAGCGATGATCGCCGCGTGGCAGGTGCGCCCGCCGCGGTTGGTGACGATGGCGCTGGCGCGCTTCATCACTGGCTCCCAGTCCGGGTCGGTCATGTCGGAGACCAGGACGTCACCCGGTTGGACCTTGTCCATTTCCGACACGTCGTTGATCACTTTGACCGGACCGGCACCGATGCGCTGGCCGATGGCACGCCCTTCCACCAGGACGGTCCCCTTCTCTTTCAGCAGGTAGCGCTCCATGACCGTGGCGCTGGCGCGGCTCTTCACGGTTTCCGGGCGTGCCTGGACGATGTACAGCTTGCCGTCGTCACCGTCCTTGGCCCATTCGATGTCCATCGGGCGGCCATAGTGCTTCTCGATGATCATGGCCTGCTTGGCCAG

Allele: 13

>trpE-GEB9ANNNTGATCGATTAANTTCCNNGNGGGATGTTCGGCGCGCAGGAAAGAGTGCCCGCCTGCAGAATTCGCCCGCAGACGCNTCCACGCGATTGAGCCCCACGCGCAATGACCCGGTGACCGTGGGCCCGATGTGCACGACGTTGGAGTAACGTTCGATCACCCATTTTTTCGGTGACCTTCACCGCGCCGAAATNGGACACCCCGCCCCACGTCGTCTGCGGCCCCCAGTCGTGAGCCGGCGTTTCGTGCCAGCTTCACCCGCGAGGACTATGAAAACGCGGTAGGAAGGATCAAGGACTACATCCTGGCCGGCGACTGCATGCAGGTGGTGCCGTCGCAGCGCATGTCCATCGAATTCAAGGCGGCGCCCATCGACCTGTACCGCGCGCTGCGCTGTTTCAATCCGACGCCCTACATGTACTTCTTCAACTTCGGCGACTTCCATGTCGTGGGCAGCTCGCCGGAGGTGCTGGTACGGGTCGAGGATGGCCTGGTGACGGTGCGCCCGATCGCCGGTACCCGTCCGCGCGGGATCAACGAAGAGGCCGACCTGGCACTGGAGCAGGATCTGCTGTCGGACGCCAAGGAGATCGCCGAGCACCTGATGCTGATCGACCTGGGGCGCAACGACGTGGGGCGGGTGTCCGATATCGGCGCGGTGAAGGTCACCGAAAAAATGGTGATCGAACGTTACTCCAACGTCATGCACATCGTGTCCAACGTCACCGGGCAATTGCGCGAGGGGCTCAGCGCGATGGACGCGCTGCGGGCGATTCTGCCGGCGGGCACTCTATCCGGCGCGCCGAAGATCCGCGCCATGGAGATCATCGACGAGCTGGAGCCGGTCAAGCGTGGAGTCTACGGCGGCGCGGTCGGCTACCTGGCATGGAACGGCAACATGGACACCGCCATTGCCATCCGCACCGCGGTGATCAAGAACGGTGAACTCCACGTGCAGGCCGGCGGCGGTATCGTTGCCGACTCGGTGCCCGCGCTGGAGTGGGAAGAAACCATCACAGCCCGCCGGGACCCGAGTCGCACGATACCGCCCCCCGGCCTGCAGTGGAAGTTCACCTTTTTGATCACCGCGATGCGGAAGCAATTGGCNNNTTCCACGTTTGNCTTTCANGNCAGTAGGCCACAGCGCCCCCTTAGAACCAGCTNGACCGCTCCNNCTTNATATAATNTCTGGGNGANATTTNCGCCGCTGANGAGTGCTCTGAAGAATCCTCATNNGACATCCGC

Allele: 7

P. aeruginosa GEB9

Allelic profile of P. aeruginosa based on sequence comparison of seven house keeping genes

Strain Acs Aro Gua Mut Nuo Pps Trp1 IISR13 15 121 36 5 4 15 82 IISR6 15 121 36 5 4 15 83 IISR51 15 121 36 5 4 15 84 IISR853 22 5 83 2 4 13 75 BP35 22 5 83 2 4 13 76 GEB9 22 5 83 2 4 13 77 PAO1 7 5 12 3 4 1 7

Population snapshot of relationship among the global P. aeruginosa-burst diagramCreated by eBurst programme which is based on a novel eBurst algorithm

IISR6 &others

BP35 & others

BP35

Dutch

Chinese Strain and

IISR51, IISR13 & IISR6

Population snapshot of relationship among the global P. aeruginosa-burst diagram Created by eBurst programme which is based on a

novel profuse algorithm

1. MLST analysis revealed that the isolates can be grouped into two

i. IISR6, IISR51 and IISR13 belong to one Sequence Type

ii. BP35, IISR853 and GEB9 belong to another Sequence Type

iii. PAO1 formed singleton in the population

2. Comparison of the allelic profile with global collection (932 isolates) revealed that the

i. IISR6, IIASR51 and IISR13 clustered with a Chinese strain (ST760) has origin in human sputum ( submitted by Yun-Song Yu Zhejiang University, PR-China, 02-06-2009)?

ii. BP35, IISR853 and GEB9 are SLV of a Dutch strain (ST 575) with unknown history (Submitted by R Van MansfieldUniversity Medical Center Utrecht, Netherlands, 14-03-2008)?

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