P‑2

Gene Engineering Division
RIKEN BioResource Research Centern RIKEN DNA BANK clone name:hmK02′ pRSETB
Rlr三 関O Clone lD

RDB 15313

O Lot

15313 A7H9O DNA Concentration

25 ng/ULO Volume40 μL

O Form

DNA solution in TE buffer

O HostDH5α

O Culture

LB medium

O Antibiotics

100 Ug/mL Ampicillin

O Purification

QIAGEN Q

Spin Miniprep kit

250 ng/、 vell″e‖
250 ngノ、

ビ ００Ш
一(NEB#N3200L),︺Ｄ Ｏ Ｃ コl kb Plus I)NA Ladder(NEB#N0472S),

一コＯ Ｃ コSuperco‖ ed DNA Ladder

一 Ｅ Ｏｍ15313

一ビ００Ш土 工Ｅｃｍ

O Digestion by restriction enzyme≡Electrophoresis :≡ ≡一 一一ロ︼ ¨ 一
一 一 一ｏお¨ｏｍ卸¨ ３ｍ８{kb)目

100 ng DNA per lane ; 1% agarose gel , 1 x TAE Buffer

Restriction enzyme

BamHI

Expected size of fragment

:

EcoRI
BarnHI+EcoRI

3.5

kbp

3.5
2.9′ 0.7kbpkbkb

O Confirmation of the insertion sequence
Please be sure to check our sequence analysis results before your request.
Sequence namePrimer lD

Primer name

Sequence - APr0261

丁7̲20bp

Confirmed feature
6xHis,T7 Tag,Xpress tag,insert 5'

Sequence - B

Pr0142flori R3

丁7 ter,insen 3'

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

APPROVED BY
http ://dna. brc. ri ken.j p

金 8醐 里ms136

28

C G A TA
T352

469

361

586

3・70

TCACCCTG■595′

487

379

604

KB 3500 P(Э P7 BDTv3.rnob

55

GGGG TTCT1‐「 7HiSQV Bases:934

Pts 1427 to 12727 1Dkl Loc:1404

NatIVersion 6.O496

388

613

163"Kθ

秩280505

2397622

172

64289

51473190「T7 ち523298

415

640

532

307

649316￨433

658

208442′
V667

325550′
V脚217

11334559

い Vi'

Electropherogram Data Page
〜1 of 2￨541′1w< t0 ].qter)tTC ACTGG TGG'C̲C C ifi■
100
109

Spacingil l.28 Pts/Pane!1350
Plate Nanne:20170908 kitaku

Sep 08,201702:17PM,JS丁Sep 08,201702:44PM,JST!nst Mode!ノ Nanne 3500/3500:nstrument

NLく=c:;TG≒ ≒ 91

L→199

T CAGGTACTACATCG ACCCCTCCGTCAATGG GCATCAGTT

ギT CATCAT CAT Cf ̲Tl GG

181

6x t'l's4()6631V｀G GGCCAACAccTTcTAccACAAGTCCAAGTTCACC GGCG TG AACTTCCCCCCCC ACGC CCCCATCATGCA GAAC

ｌ︲＝ Ｖ577

い ⅣVI

VWI ザ咽424

TGG CCCAGGGCCGGCCAATCCCTTTCGCCTTCGACC TGG TGTCCCTG AGCGTG ATCAAGCCCC AGATGAAGハ46 .,ATF

54IC TC

D04498J4̲A7H9̲1̲T7̲20bp̲E08̲14̲AB124
D04498J4̲A7H9̲1̲T7̲20bp

G A C C37

primer name A :T7′ Obp
TAATACGACTCACTATAGGG‑3:5'‐19

NNNNNT T T■ GT Tさ ヽN CT N NNN N■127:G̲T C T G T A C G A C G A T G10N Nヽ NN

N(3:199A:274T:158C:274

鯰 憮
SノCap:14KB.bcp
KB l.4.1.8
さNNNNl「 GG1 18￨￨〉

―・―:一3413

460

CGGCG GC TCCCCCTCCGTG ACC GCCCiC■451568

11′11

CAGAGCGTGGACTGGGACCCCTCCACCGAGAACATCACCCCCAGCGACGGCGTGCTCAACGCCGACGTGACCATGTACCTGAACCTGG2,GGGCCCCGIGCAACCACAAGT478

CCTGTTCTGTTACG GCCACAG AGTTTTTACCAACTACCCACAAGAGATCCCAG ACTATT TcAACCACCCCTT TccTcA̲GGCCCTG TCCTGGGAGACGTCCCTGCAGTTCGAGG]CACAATCGACGGTCACCCCACAGGCACACCTTACGACGGACATCAGGACATGACACTGCCCCTCACAA
226235244253262271→

G Tl CGG

NN

1308654013086540130865401308
65401308
65401308
6540

Printed on:丁 ue Feb 20,202402:16PM,JST『いｖＷｖ

SノApp:iedBibζ ystems

Cap:14

NG:199A:274丁 :158 Ci27473685

892

Tム ACC

694982

lNN

1072

991

NNN N NN

113

N NNN

910

1000

NN N
1081

D04498J4̲A7H9̲1̲T7̲20bp̲E08̲14̲AB:24856

lnst Modelノ Name 3500/3500 instrument

757

G NCC(3GGC‐865NNNNGG NG T

1045

NNC TC

877751054

N NN N

NC N NN964

NNll17NN N N

9・Electropherogram Data Page 2 ol 2

NN

NN NNN N NN N N NN

955

NSep 08,201702:44PM,JST

Sep 08,201702:17PM,JST

7391036

l108

NN N

NNN NN NNNN

946

ANGNN NTATr.TCC

GCTG N TTGGC TCCTCCC CCGC NN

748

D04498J4̲A7H9̲1̲丁 7̲20bp
KB 3500 P(Э P7 BDTv3.rnob

GGCTCGTG NG‐

P:ate Nanne:201 70908 kitaku

730

G CCCC

937■ AGG

N'NNGGN

NN N N N N

1027

N1099

N838

‐C.CTICiTCC GCC C・Spacingil l.28 Ptsノ Pane!1350

721
1̲̲ゝ928

1018

NN N

847

HiSQV Bases:934

GG

Pts 1427 to 12727 Pkl Loc:1404

Version 6.O

820

919

1009

NNN1090

N N NN NN

N NNCNlGG GG TTTTTNCC TG■

GCTIGNTCCGGCTCCTN C

712

NN

829

(3iTCCT CG■ C‑lTGCC C(〕 C CG

811‐
G NT TC G

秩ぃにθ2703

Ci「 .GG CCCCCiriGG■

802901

CCTTGGGGCC TCTNAACGGG NCTNG「793

NG CGCCGTC GCCC CTCCT784

Cた1063l126

NNNN

NNNN N NN NN NN N

NN NNNN NNGNCN N Cr̲NNN N TNN NCN NN NCN NN NNN NNNN N N CNN NCNNN N‐883、TiiCT=lG〕 NGTGG

676

CCi GF‐ TG■ .― GiCCiC CT■

KB l.4.1.8

KB.bcp1308
65401308
65401308
65401308
65401308
65401308
654
Printed on:Tue Feb 20,202402:16PM,JSTSノApp‖ edBibζ ysterns

NG:48A:54T:54C:76
Cap:17

RI

28‐￨:flo「 :̲ 3137

D04498J4A7H9 1 flori R3 F08 17 AB!24
R3
D04498J4A7H9 1 flo
KB̲3500̲POP7̲BDTv3.ITlob

64聰 :i網 :17麗 計 :患 !:::46

163172

181WIWITCC CG■ T'T∴ GTTCCTCCTT TCiC
738291

190

199379

604

271

496

■GTTCAC

280

397

289

406

298415307424316433

ACTTGGACTTGTCGT■GAAGGTGTT GCCCCTCiGGCTG■
514
523
532
541

622

631

640

649

i

21,1「r、.∩■υ υ

Ⅲ
…
…
…
…

217

334

HMよ■.こ のRl

325

442

f

109工 V´

667

226

GTC(GACCCC

G‐

6'7 6'\.AACTCTGTGGC(TGT GGGcCcTCAC CCAGGC(5591451‐ブ̲T71●ψι↑ヽ■。
r1ic̲「 ■̲:̲ICCCCTC…

Spacing:11.45 Pts/Pane11 350
P!ate Narnei 20170908 kitaku

Sep 08,201702:17PM,JS丁Sep 08,201702:44PM,JST

lnst Modelノ Narne 3500/3500!nstrument

658

CCTGC TTGAAATAGTCT GGGATC TCT TCTCG GTACTTC GTAF

Electropherogram Data Page 1 of 2550

TGGTCACGTCCCCCTTCACCACCCCGTCGCTGGCGGTcATCTTCTCCGTGGAGGGCTCCCiCT(CC CG GTG
505613…

CTC■ GC TTCCTTTCGGGCTTTGTTAGCAGCCGGiTCAACCTTCCAATTc

NCC(〕 TC CG GG CCTC TTCG CT■

154

ＨＶ145

TTATTCCTCACCG GTG GCAGCAGCCFi■262487388TGTTGCCCTCCGTCTTCCTCACCAGCCTGTGGCCCATGTAGTCGTCCCCGGGCATCTCCACGATCTCCTTGCCGGccTTCTAGGT

370595

ＷＶ＝ＶＩ

586

哺Ｙ

577＝Ｖ136●253

こNNN TTG GGlNNGGCGヽ

TTMTGCGこ CGCWAGGGC‑311

pr:mer narne
5:‐

NNNNN NN NNNヽ ヽNN NNN]NN
1019127

Tγ 寸々r″ 、lИ α

244

478

361

352

469343460

TGr、 T CCGGCCGTCG GCG GGG■

568

3GAGCCCCC(3TCCTC CAACTCCACGCAC CTCTCCCAGG ACAGGCCCTCAGG AAACG・￨￨3CiCGCTCTGGTTCTGCGGTCTTCATCTGGCACTTGTCGTTGCCGCCGCCCTCCAGC TTCAG GTAC■

235]GCCACG GCGTCCTCCACCTCCTCGGTG■

l18

TTTAG■ GCC CCCA■ G GGGTTATGCl

NさKB.bcp
KB l.4.1.8

1280
64001280
64001280
64001280
64001280
64001280
640
0Printed oni Tue Feb 20,202402:16PM,JSTApp:iedBibζ ystems

685

λぃ￨(02694712

793
802
811

1009

820

:nst Mode!ノ Name 3500/3500:nstrument

Sep 08,201702:17PM,JST
Sep 08,201702:44PM,JST

D04498J4A7H9 1 flori R3 F08 17 AB!24
D04498J4A7H9 1 flori R3829

730

838

937

739

N NN

N9461036l108

NNN NNNN856757

955

1045

NN N

NN

865

TG

964TCN

1054

ll17

N N N

874775973

NCNNN N NT31063

1Electropherogram Data Page 2 of 2

N lN N NN N

NNN N NN N N NNNNN N NNNNN NNN N N NN NNN NN

847

lC NNG TC｀ .TCCNAN CNN Tメ
928

1099

NN

NIN N N N N
102

さNN

Plate Narne:201 70908 kitaku

KB 3500 P(Э P7 BDTv3.rnob

1018

N

NNNTT N NNNNG TGTC TCTCCTGrTGTCCCTCG T「Spacing:11.45 Ptsノ Pane11 350

748

Pts 1420 to 12780,Pkl Loc:1397721

Version 6.O HiSQV Bases:942
GCG■ ■「 G GC TTCGCCCCCCCTCCGCC TTGTC CGCC C‐← 一⊥ ̲703

iCAG■ ACACGTCG GACACCAC GTCCAA

Cap:17

S/NG:48A:54丁 :54C:76

14901

NN■N NN N NNN
1081

NN NC■ GATN CN CiNC NNT TTN CTCNNC「919910892

TT■ TCG NCNTC GTC GT■NNNNN Nさ1000

NN NNN N NN

991

TCT NNNNNNT N

N NN NN N1072

NNN NN N N N

982

T CN TNN

26

NN N NNIN NN TG ANG riT G:=T

3TGCCCTC CCCTCG TTCTC̲CTC TCC CC,TTC CGG NCCG TCC TGT GT CCTC TC TTC TC TCGGGCTTG TC CCC TCNCN N(3G NCGG〕GTAKB.bcp
KB l.4.1.8

128064001280
64001280
64001280
64001280
64001280
640
Printed oni Tue Feb 20,202402:16PM,JS丁