P‑4Gene Engineering Division
RIKEN BioResource Research Centern RIKEN DNA BANKclone name : pUG18-HTK (with promoter)

O Clone lD

RDB 03436

O Lot13745 A7KaO DNA Concentration

25 nanogram/microliter

O Volume

40 microliter

O Form

DNA solution in TE burer

O HostDH5 alpha

O Culture

LB medium

O Antibiotics

25 microgram/ml Kanamycin

O Purification

QIAGEN QlAprep Spin Miniprep kit

O Digestion by restriction enzyme∞∽ ＋Ｈ一03436

l kb Pius DNA Ladder〇ｍ一(kblＨ一０ ∽250 ngノ 、vell

目 一Ｏ ｍ一コ Ｏ Ｃ コ(NEB#N3200L),

6.0●●●・・ 口・ロニ
薔 言會

Electrophoresis i

100nanogram [)INA perlane i ll%agarose gel,lx・ T′へE Burer

Restriction enzyrne

Expected size of fragnnent:

Bg:=工3.7kbp

3.7
3.■ ′0.6

Sal=

Bg:==+Sal=kbp
kbp
kbp

O Confirmation of the insertion sequence
Please be sure to check our sequence analysis results before your request.
Sequencg naJne

Primer lD

Primer name

Confirmed feature

Sequence - A

Pr0324

AFRinsert 5'( pro,HTK 5')

Sequence - B

Pr031 0pUC― M13̲‑100

insert 3'(HTK 3')

Sequence - C
Sequence - D
Sequence - E
Sequence - FヽSequence - G
Sequence - H

APPROVED BY
http://dna. brc. riken.jp

バBttB瑚 見ms
S/N G:141 A:194 T:153 C:183 p6rner name

KB.bcp1019

A : AFR
2837′

352

577

olへ 1

361

586

478

145370595

May 17,201903:15PM,JS丁:nst Modelノ Nanne 3500′ 3500 1nstrument
iV:ay 17,201902:33PM,JS丁AGACATGCAGGGTTTTCCTGCATTTTTCTTT

レ.十 ″

73181

406

298415A307

t toι514

91

316

631

523

640

532

649

433

658

100

550334A559

C7,"i109

GTCCACTC217667442

卜IT Iこ /325

AAAG GACCAATAATAATGACTAGA

208

CCACGGTCCCCAAACGGATcCCTCAG199424

541

AACGCAATTCGTGTTTATGGcTCTCTTGGTCGTCACACTCAT…

づ久λI

82

Plate NarTle:201 9051 7 kitaku
64

Prク

βTGGTACAATGAACATCC

Spacing:12.64 Ptsノ Pane!1350

50172

397

190

HiSQV Bases:955

163505622

6‐45 1

Pts 1462 to 13985 Pkl Loci1439

03436̲13745̲A9匡 gF̲AFR
KB 3500 P(DP7 BDTv3.rnob

03436̲13745̲A9EgF̲AFR̲D04̲04̲AB:08154379

496

613

Electropherogram Data Page 1 of 2

CACACGGAGTACAGTTCACCTATCAATC GACAAC CG GTCAGTCCAAGCCG GAAGTCAATTTTTATA CC CAAGAGATT
487604

VVVl「ATCTCGGGTGGAACCGGATTGGCCCCTTACACATGGTCCATTTTTCTCTATTTTGCCGATTTATGATCCAGGTGGATAcTTTCACAAAGTGTACCAAAC

469289T GATCAAAC GACCAATCACCA̲TCCCAATCG AC GGAccCCCCCCGIGCCGGG AA̲AA′136

Ａ﹁127′343

AACAAAGAATCAACATTGTTCATCAAATTAAGGAACGAATATTCCATAAATATGGGGATGATGTT460

TATTC GGiTATTGACATGATGTIGTGTTCTGTCArゝ568

ACTAGAT TAT CC388

GCCTTTCCTACGTATACAT CGATACCGGCGCCATGTATCGCGCCCTGACGTACCCGGCCCTTGCATGCGGC
2623524425326227128o118

ATCAAGC GATTTTCG GTAAAGC TACG AGGAAハ15'‐ GGAATTGTGAGCGGATAACAA‑3'
Version 6.o
KB l.4.1.8 Cap:4
NNN NN Nヽ N lヽ NN N INNT G さN NNN Gr、 さNT CCAGcTCC GTFCCCGTTGiC

1216
60801216
60801216
60801216
60801216
60801216
608
Printed on:VVed Nov 24,202112:09PM,JST

ＶＹ

Cap:4

NG:141A:194T:153C:183

バB金 瑚繋ms
SノKB.bcp

76685694703712

May 17,201902:33PM,JST!nst Modelノ Nanne 3500/3500 1nstrument

iVlay 1 7,201903:15PM,JST03436̲13745̲A9匡 gF̲AFR̲D04̲04̲AB10803436̲13745̲A9匡 gF̲AFR
KB 3500 P(Э P7 BDTv3.rnob

P:ate NalTle:201 9051 7 kitaku

Spacing:12.64 Ptsノ Pane11 350

一811

919

1018

820

1099

829

937

NTGTGCC「1027〕
N、NC

1036

946

739

847

748

856

N

NNN NN NNN

955

ll17

NN N NNN N NN NN NN N

1045

TCGNANTGTCTGGNCNNCTTTNCGANCN

838l108

N NNN N NN

N NNNN N NNCNNN N N NN NN

N N

757865

N

9737668749828831072l135

N

NN N:NNNN NN NNGCi

N N N NNN N

1063

N N NNN NNNCN N NNNN N NN NN

1054

N NN

l126

NN991

784

HiSQV Basesi 955
739
ヽプ802910

1009

1090

lN NN

N NN NN NNNN NNN1153

Electropherogram Data Page 2 ol 2

N NN

Pts 1462 to 13985 Pkl Loc:1439

Version 6.O
721︲︲一793901

GC■ NTT■ ・、NCAACCANNTCTTCCTCCFiCGTTATGTCCFi■1000

N N

N N N N N

NN N N NN N
1081

NNl144928CAACATTTCTACNATCCTTCACTGTACACGTGGCAATCNCiNGTCCCATGTTCATTCGTCTCCATCATCCCATCTGTTATACNACG AGCGCTTCNGTCTTAAC

2N'N NN NTT C T C GNN NGGGGN NN C N

ヾ NN〕

ヾNN964TGCTAAATCGGTAGAAGCCCAAAAGTTCCACGATGCGATCTGTGCCCTTATCGTCIIAAGAGCTGTTTGAATATGcAGGcAAATGGcGTATTATTcGTGTGcAA

KB l.4.1.8

1216
608
01216
608
01216
608
01216
608
01216
608
01216
608
Printed on:VVed Nov 24,2021 12:09PM,JSTバB合 8馴里ms5'―

121131

ら:puc̲M13‐

‖Tに

141

241

251

31

8寵

1 KB̲o500̲POP7̲BDTv3.mob

03436̲13745̲Bl KoS̲pUC‐ M13̲‐ 10003436̲13745̲BlKoS̲pUC― M13̲‐ 100̲F02̲17̲AB:24

100

261

151

41271

51ヲ 2■

iCG
57351

581

361

591

371γヽ
7Jへ′―Ⅵ

601

r′I

381

VIVヽ

611

391

「VV
CATGTGTAACCGIGCCAATCCGGTTCCACCCCANNNCCATAATCTAGTANAATCTCNTCC

103:37PM,JST

281

61

融 hこ L

171

621

401

291

・

71

301

CT TN

411

631

91

321

211

101

331

221

56]

4

Plate Nannei 20211125 dnabank

Spacing:10.61 Pts/Pane11 350

Nov 25,202102:33PM,JSTNov 25,202103:01PM,JST!nst Modelノ Narne 3500′ 3500 1nstrument

201

GC CATTCCAGAAGTTTcTCACGGTCGG

311

551

NN

G C TC G T CGT AT AFI,CACATG CC ATC AT

541

661

GATCATAAATCGGCAAAATAGAICGCCr、 TTTCCCTCCATATTCAAACAG CT
441
43181421

VWV｀641￨Electropherogram Data Page 1 of 2

NINT c .CTT CN CCC TT

531

191

CT GCAGCGTAACCAACATC ATTAACAATTATTAG ACGTCAT亀+■181521lGGTTGCTTAACTCCTTCAGTTAAGACCGAACC
4=⊆ ̲全 ユL⊆ ¨二161

AAAATGGTATGCGT TTTCACACATCCACTATATATCCGTG TCG TTCCGCCCACT CCTC AACCCCATTCCACAAATTCTC TA

11N NNNlN CNN N NN NTTT

Cap:1 7T

ATTACGC&Gこ TGGCGIMGG‑3'漁‖ :『肌 罠 :ょ 聯
G T CAC G ACG T TG TAAAACC ACC C CCAC TG CCAACC TT C CATGTS/N(3:1310A:1324 丁:1223C:1,Primer name

1CGG

TTCTACGATAACCCCACAGATCCCATCGTCGAACTTTTGGGCTTCTACCCATTTACCAGTTTCGTACACTTTCTCAAACTATCCACCTG
51461471481491501511341

CAGACCAATCAACATCGCACCTCCCATTGCCACCTGTACAGTCAACGATGGTACAAATGTTGTCGGTCCTTGCACACGAATATTA231

AAGTT GACCAC ACATTACGAC CTGGCACAGTTGGACATAACCTGG ACGハ11N NNNKB.bcp
KB l.4.1.812226110122261101222611012226110122261101222611
Printed on: Thu Nov