pET-15b Vector

TB045 12/98
The pET-15b vector (Cat. No. 69661-3) carries an N-terminal HisTag sequence followed by
a thrombin site and three cloning sites. Unique sites are shown on the circle map. Note that the
sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the
circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below.

pET-15b sequence landmarks

453-469
452
362-380

EcoR I(5706)
Cla I(24)
Hind III(29)

Aat II(5635)
Ssp I(5517)

319-335
213-259
(866-1945)
3882
4643-5500

Sca I(5193)
Pvu I(5083)
Pst I(4958)

Ap
(4
6

T7 promoter
T7 transcription start
HisTag coding sequence
Multiple cloning sites
(Nde I - BamH I)
T7 terminator
lacI coding sequence
pBR322 origin
bla coding sequence

Bsa I(4774)
Ahd I(4713)

)
00
55
43

Bpu1102 I(267)
BamH I(319)
Xho I(324)
Nde I(331)
Nco I(389)
Xba I(428)
Bgl II(494)
SgrA I(535)
Sph I(691)
EcoN I(751)

866-1945)
lacI (

pET-15b

AlwN I(4236)

(5708bp)

Mlu I(1216)
Bcl I(1230)
BstE II(1397)
Apa I(1427)

o ri (

BssH II(1627)

38

Hpa I(1722)

82
)

BspLU11 I(3820)
Sap I(3704)
Bst1107 I(3591)
Acc I(3590)
BsaA I(3572)
Tth111 I(3565)

PshA I(2061)

Bsm I(2704)
Msc I(2791)

Bpu10 I(2926)

Eag I(2284)
Nru I(2319)
BspM I(2399)

T7 promoter primer #69348-3

Bgl II

T7 promoter

lac operator

Xba I

rbs

AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGA
Nco I

HisTag

Nde I

Xho I BamH I

TATACCATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGGCCTGGTGCCGCGCGGCAGCCATATGCTCGAGGATCCGGCTGCTAACAAAGCCCGA
MetGlySerSerHisHisHisHisHisHisSerSerGlyLeuValProArgGlySerHisMetLeuGluAspProAlaAlaAsnLysAlaArg
Bpu1102 I

thrombin

T7 terminator

AAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG
LysGluAlaGluLeuAlaAlaAlaThrAlaGluGlnEnd
T7 terminator primer #69337-3

pET-15b cloning/expression region

Novagen ORDERING 800-526-7319

TECHNICAL SUPPORT

800-207-0144

pET-15b Restriction Sites

Enzyme
AatII
AccI
AceIII

# Sites
1
1
7

AciI
AflIII
AluI
AlwI
Alw21I

91
2
24
16
8

Alw44I
AlwNI
ApaI
ApaBI
ApoI
AvaI
AvaII

4
1
1
2
2
2
9

BamHI
BanI
BanII
BbsI
BbvI
BccI
Bce83I

1
13
3
5
31
16
7

BcefI
BcgI
3431
BclI
BfaI

5
8
5218
1
6

BglI
BglII
BmgI
BpmI

3
1
1
6

Bpu10I
Bpu1102I
BsaI
BsaAI
BsaBI
BsaHI

1
1
1
1
3
8

BsaJI
BsaWI

11
7

BsaXI
BsbI
BscGI
BsgI
BsiI
BsiEI

1
2
13
3
3
6

BslI
BsmI
BsmAI

22
1
7

BsmBI
BsmFI
BsoFI
Bsp24I
Bsp1286I
BspEI
BspGI
BspLU11I
BspMI
BsrI
BsrBI
BsrDI
BsrFI

2
4
57
12
11
2
3
1
1
25
3
4
8

Locations
5635
3590
983
1711
3772 5012
1216

3820

716
4138
1196
4236
1427
900
1491
324
1768
2826
319

1200
5299
3634

600
1362

208
4450
735
1508
5252
1230
257
4903
2280
494
1425
1054
4783
2926
267
4774
3572
493
539
2551

2042

2523
5384
4134

2397
5706
2770
2144
3105

2232
4851

614
1701

1427
2075

2030
5318
1076
1542

2814

3470

3638

5380

2481
5073

2938

2784

5691

2200

3911

4209

1703
2042

2512
2076

4322
3397

429

2834

2514

4833

1543

3329

2177

4315

2731

4568

3017
674
5632

1173

1856

189
4173
1875
3536

1535
5004

2038

3009

4026

1067
3993
2001
5232

1267
5377
2287

2704
913
4774
1831
677

5256

1318
5550
3461
2218

1444

2443

189
2404
3820
2399

3009
2481

3346

449
1263
160
2274

3753
1629
526
2628

5554
4774
535
4793

4160

1831

5083

# Sites
1
1
1
3
11
41
25
26
1
97
26
11
29
3
2
1
3
6

EagI
Eam1105I
EarI
EciI
Eco47III
Eco57I
EcoNI
EcoO109I
EcoRI
EcoRII
EcoRV
FauI
FokI
FspI
GdiII
HaeI

1
1
3
5
3
2
1
5
1
11
2
18
14
3
5
8

HaeII
HaeIII
HgaI
HgiEII
HhaI
Hin4I
HincII
HindIII
HinfI
HpaI
HphI
MaeII
MaeIII
MboII
MluI
MmeI
MnlI
MscI
MseI
MslI

13
28
15
2
45
5
2
1
14
1
17
12
18
15
1
2
34
1
24
10

MspI
MspA1I
MwoI
NarI
NciI
NcoI
NdeI
NgoAIV
NlaIII
NlaIV
NruI
NspI
Pfl1108I
PflMI

35
11
44
5
14
1
1
4
31
29
1
4
2
3

3461

3091

4948
902

Enzyme
BssHII
Bst1107I
BstEII
BstXI
BstYI
Cac8I
CjeI
CjePI
ClaI
CviJI
CviRI
DdeI
DpnI
DraI
DrdI
DrdII
DsaI
EaeI

3347

499
560
5250

2980
5684
3736

TB045 12/98

2114

Locations
1627
3591
1397
1018 1147

Enzyme
PleI

1270

24

4579
3513
939
389
524
5101
2284
4713
834
993
621
4368
751
240
5706

4598
3928

5290

653
656

2792
1890

3704
2740
2122
5380

5508
3894
3074

649

2784

187

1666

2703
524
944
3835

2801
656
2265
3846

814

4406

16
1722
29

1115
5254

4935
1890
2337
4298

2486

2284

4040

2826

2284
2394

4712

2789

4868

5689

5101
2791

4786

1722

1216
4035

# Sites Locations
7
477
765
4199 4702
PshAI
1
2061
Psp5II
2
2784 2826
Psp1406I 5
878
2246
PstI
1
4958
PvuI
1
5083
PvuII
3
1816 1909
RcaI
4
614
4540
RsaI
4
165
1363
SapI
1
3704
Sau96I
21
Sau3AI
29
ScaI
1
5193
ScrFI
25
SfaNI
24
SfcI
5
138
462
SgrAI
1
535
SphI
1
691
SspI
1
5517
StyI
3
244
389
TaqI
13
TaqII
8
1124 1342
5246
5399
5416
TfiI
7
1895 2197
3374 3795
ThaI
41
TseI
31
Tsp45I
9
124
1397
3472 3567
Tsp509I 16
Tth111I
1
3565
Tth111II 7
1055 1748
4449 5705
UbaJI
26
VspI
4
477
1901
XbaI
1
428
XcmI
3
1072 1588
XhoI
1
324
XmnI
2
3378 5312
Enzymes that do not cut pET-15b:
AflII
AgeI
AscI
BseRI
BsrGI
Bsu36I
KpnI
MunI
NheI
NspV
PacI
PmeI
RsrII
SacI
SacII
SfiI
SgfI
SmaI
SrfI
Sse8387I StuI

4219

2791
1268
3002

1556
3393

1586
4965

2376
5124

2807
5483

539

560

674

1856

2551

389
331
526

2114

2274

2628

2319
691
2103
798

3165
4731
2666

3457

3824

2715

Novagen FAX 608-238-1388 E-MAIL novatech@novagen.com

852

1648

3714

3145

4939

5312

3411
5548
3626

5653
5193

4085

4276

4954

2015

3722

5061

2351

2649

2870

2225
4969

2492
5180

3259

3281

4410

4417

1960

4885

2714

1606

AvrII
DraIII
NotI
PmlI
SalI
SnaBI
SunI

BaeI
FseI
NsiI
RleAI
SexAI
SpeI
SwaI