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pET28a-c(+)质粒图谱

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2024年4月15日发(作者:蓝高超)

pET-28a-c(+) Vectors

pET-28a DNA

pET-28b DNA

pET-28c DNA

Cat. No.

69864-3

69865-3

69866-3

TB074 12/98

The pET-28a-c(+) vectors carry an N-terminal His•Tag

®

/thrombin/T7•Tag

®

configuration plus

an optional C-terminal His•Tag sequence. 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 poly-

merase is shown below. The f1 origin is oriented so that infection with helper phage will produce

virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-

stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).

Call 137-649-32037 to buy the vector in China.

pET-28a(+) sequence landmarks

Pvu I

(4426)

Sgf I

(4426)

Sma I

(4300)

K

a

n

(

3

9

9

5

-

4

8

0

7

)

T7 promoter

T7 transcription start

His•Tag coding sequence

T7•Tag coding sequence

Multiple cloning sites

(BamH I - XhoI)

His•Tag coding sequence

T7 terminator

lacIcoding sequence

pBR322 origin

Kan coding sequence

f1 origin

370-386

369

270-287

207-239

Dra III

(5127)

Bpu1102 I

(80)

Xho I

(158)

Not I

(166)

Eag I

(166)

Hind III

(173)

Sal I

(179)

Sac I

(190)

EcoR I

(192)

BamH I

(198)

Nhe I

(231)

Nde I

(238)

Nco I

(296)

Xba I

(335)

Bgl II

(401)

SgrA I

(442)

Sph I

(598)

158-203

140-157

26-72

773-1852

3286

3995-4807

4903-5358

0

3

-

5

3

5

8

)

(

4

9

n

i

g

o

r

i

1

f

The maps for pET-28b(+) and pET-28c(+)

are the same as pET-28a(+) (shown) with

the following exceptions: pET-28b(+) is a

5368bp plasmid; subtract 1bp from each site

beyond BamH I at 198. pET-28c(+) is a

5367bp plasmid; subtract 2bp from each site

beyond BamH I at 198.

Mlu I

(1123)

Bcl I

(1137)

BstE II

(1304)

Apa I

(1334)

BssH II

(1534)

EcoR V

(1573)

Hpa I

(1629)

7

3

-

1

8

5

2

)

l

a

c

I

(

7

Cla I

(4117)

Nru I

(4083)

pET-28a(+)

(5369bp)

Eco57 I

(3772)

AlwN I

(3640)

(

3

i

o

r

BssS I

(3397)

2

8

6

)

PshA I

(1968)

Bgl I

(2187)

Fsp I

(2205)

Psp5 II

(2230)

BspLU11 I

(3224)

Sap I

(3108)

Bst1107 I

(2995)

Tth111 I

(2969)

T7 promoter primer #69348-3

pET upstream primer #69214-3

T7 promoter

lac operator

rbs

Bgl

II

Xba

I

AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGA

Nde

I

Nhe

I

T7•Tag

His•Tag

Nco

I

TATACCATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGGCCTGGTGCCGCGCGGCAGCCATATGGCTAGCATGACTGGTGGACAGCAA

MetGlySerSerHisHisHisHisHisHisSerSerGlyLeuValProArgGlySerHisMetAlaSerMetThrGlyGlyGlnGln

thrombin

Eag

I

His•Tag

Bam

H I

Eco

R I

Sac

I

Not

I

Xho

I

Sal

I

Hin

d III

ATGGGTCGCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCC pET-28a(+)

MetGlyArgGlySerGluPheGluLeuArgArgGlnAlaCysGlyArgThrArgAlaProProProProProLeuArgSerGlyCysEnd

...GGTCGGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCC pET-28b(+)

...GlyArgAspProAsnSerSerSerValAspLysLeuAlaAlaAlaLeuGluHisHisHisHisHisHisEnd

...GGTCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCC pET-28c(+)

...

T7 terminator

Bpu

1102 I

GAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG

T7 terminator primer #69337-3

pET-28a-c(+) cloning/expression region

Novagen

ORDERING

800-526-7319

TECHNICALSUPPORT

800-207-0144

pET-28a(+) Restriction Sites

Enzyme

AccI

AceIII

# SitesLocations

21802994

78901618

31764967

AciI77

AflIII211233224

AluI22

AlwI13

Alw21I7159190

30423542

Alw44I311033038

AlwNI13640

ApaI11334

ApaBI1807

ApoI61921398

4940

AvaI21584298

AvaII516752051

BamHI1198

BanI9253445

17621892

BanII6190507

5202

BbsI412691608

BbvI27

BccI14

Bce83I6211937

3854

BcefI6642983

5153

BcgI9160194

19491983

BclI11137

BfaI770232

40265278

BglI12187

BglII1401

BmgI11332

BpmI49611450

Bpu10I223304443

Bpu1102I180

BsaAI229765127

BsaBI3400406

BsaHI5446467

BsaJI1057296

21963384

BsaWI721442

35774561

BsaXI217825075

BsbI229405034

BscGI11

BsgI39741174

BsiI13397

BsiEI51691908

BslI23

BsmI243104387

BsmAI68201225

4442

BsmBI317382865

BsmFI45842125

BsoFI48

Bsp24I12

Bsp1286I12

BspEI222413

BspGI12750

BspLU11I13224

BsrI22

BsrBI43563157

BsrDI211701536

BsrFI7433442

43805228

BssHII11534

Bst1107I12995

4

Enzyme

BstEII

BstXI

BstYI

Cac8I

CjeI

CjePI

ClaI

CviJI

CviRI

DdeI

DpnI

DraIII

DrdI

DrdII

DsaI

EaeI

EagI

EarI

EciI

Eco47III

Eco57I

EcoNI

EcoO109I

EcoRI

EcoRII

EcoRV

FauI

FokI

FspI

GdiII

HaeI

HaeII

HaeIII

HgaI

HgiEII

HhaI

Hin4I

HincII

HindIII

HinfI

HpaI

HphI

MaeII

MaeIII

MboII

MluI

MmeI

MnlI

MseI

MslI

MspI

MspA1I

5342

MwoI

NarI

NciI

NcoI

NdeI

NgoAIV

NheI

NlaIII

NlaIV

NotI

NruI

NsiI

NspI

39

4

12

1

1

4

1

26

22

1

1

2

4

# Sites

1

3

9

40

26

30

1

86

22

11

21

1

3

2

3

4

1

3

3

3

1

2

3

1

10

1

17

9

1

4

6

14

24

11

2

47

3

2

1

18

1

16

14

16

12

1

7

25

25

6

29

9

Locations

1304

9251054

132198

24163865

1177

401

3876

687

4675

1899

Enzyme

Pfl1108I

PflMI

PleI

PshAI

Psp5II

Psp1406I

PvuI

PvuII

RcaI

RsaI

SacI

SalI

SapI

Sau96I

Sau3AI

ScrFI

SfaNI

SfcI

SgfI

SgrAI

SmaI

SphI

SspI

StyI

TaqI

TaqII

TfiI

1169

2769

2205

166

851

4513

1178

2910

431

2172

2443

4064

563

3239

2505

4670

1797

3250

2583

ThaI

TseI

Tsp45I

3702

Tsp509I

Tth111I

Tth111II

3810

4112

1629

4654

UbaJI

VspI

XbaI

XcmI

XhoI

XmnI

20

1

8

21

5

1

3

1

2

38

27

7

# Sites

1

2

9

1

1

4

1

3

3

3

1

1

1

14

21

22

23

4

1

1

1

1

2

2

15

6

9

Locations

2010

7054689

384672

36034658

1968

2230

7852153

4426

17231816

5213944

12703030

190

179

3108

759

5062

1555

5070

TB074 12/98

3118

2549

2815

4819

4261

4912

623

3538

11072218

4117

4

2139

466

2018

521

1982

2230

580

5164

1334

2342

2509

1043

4081

2107

1610

228

2801

336

3315

3726

1415

2835

2238

3613

4745

1449

5127

2917

846

296

166

166

741

900

528

3772

658

53

192

256

3250

1573

3332

5132

560

431

3108

3298

2029

4338

556

846

3371

5082

2196

563

4239

3444

2478

1797

2230

1161

3384

1701

4314

1758

4671

369

4426

442

4300

598

4351

57

1031

5031

1802

4337

348936805346

4919

296

1249

2104

4393

1922

2274

4565

3126

2778

4656

4680

3199

3719

1304

4573

2969

962

3853

384

335

979

158

2782

2132

5300

266328762971

20842751

1655

4262

1808

1495

4815

2685

4389

1867

1513

38143821

721

1022

181

173

1629

46254814

2421

581

560

4297

1945

1080

566

4298

2413

1763

1758

4699

3430

2384

3

1123

3439

4633

3623

5104

4

Enzymes that do not cut pET28a(+):

AatIIAflIIAgeI

BaeIBsaIBseRI

Bsu36IDraIEam1105I

MscIMunINspV

PmlIPstIRleAI

ScaISexAISfiI

SrfISse8387IStuI

AscI

BspMI

FseI

PacI

RsrII

SnaBI

SunI

AvrII

BsrGI

KpnI

PmeI

SacII

SpeI

SwaI

1351

4442

2495

17382865

1175

2797

84

2815

446

296

238

433

231

12406

264

2934

467

1153

3566

581

1723

3811

1763

1816

2

4825

809

5271

20212181

166

4083

4276

598

4542

256928613228

Novagen

FAX

608-238-1388

E

-

MAIL

novatech@

2024年4月15日发(作者:蓝高超)

pET-28a-c(+) Vectors

pET-28a DNA

pET-28b DNA

pET-28c DNA

Cat. No.

69864-3

69865-3

69866-3

TB074 12/98

The pET-28a-c(+) vectors carry an N-terminal His•Tag

®

/thrombin/T7•Tag

®

configuration plus

an optional C-terminal His•Tag sequence. 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 poly-

merase is shown below. The f1 origin is oriented so that infection with helper phage will produce

virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-

stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).

Call 137-649-32037 to buy the vector in China.

pET-28a(+) sequence landmarks

Pvu I

(4426)

Sgf I

(4426)

Sma I

(4300)

K

a

n

(

3

9

9

5

-

4

8

0

7

)

T7 promoter

T7 transcription start

His•Tag coding sequence

T7•Tag coding sequence

Multiple cloning sites

(BamH I - XhoI)

His•Tag coding sequence

T7 terminator

lacIcoding sequence

pBR322 origin

Kan coding sequence

f1 origin

370-386

369

270-287

207-239

Dra III

(5127)

Bpu1102 I

(80)

Xho I

(158)

Not I

(166)

Eag I

(166)

Hind III

(173)

Sal I

(179)

Sac I

(190)

EcoR I

(192)

BamH I

(198)

Nhe I

(231)

Nde I

(238)

Nco I

(296)

Xba I

(335)

Bgl II

(401)

SgrA I

(442)

Sph I

(598)

158-203

140-157

26-72

773-1852

3286

3995-4807

4903-5358

0

3

-

5

3

5

8

)

(

4

9

n

i

g

o

r

i

1

f

The maps for pET-28b(+) and pET-28c(+)

are the same as pET-28a(+) (shown) with

the following exceptions: pET-28b(+) is a

5368bp plasmid; subtract 1bp from each site

beyond BamH I at 198. pET-28c(+) is a

5367bp plasmid; subtract 2bp from each site

beyond BamH I at 198.

Mlu I

(1123)

Bcl I

(1137)

BstE II

(1304)

Apa I

(1334)

BssH II

(1534)

EcoR V

(1573)

Hpa I

(1629)

7

3

-

1

8

5

2

)

l

a

c

I

(

7

Cla I

(4117)

Nru I

(4083)

pET-28a(+)

(5369bp)

Eco57 I

(3772)

AlwN I

(3640)

(

3

i

o

r

BssS I

(3397)

2

8

6

)

PshA I

(1968)

Bgl I

(2187)

Fsp I

(2205)

Psp5 II

(2230)

BspLU11 I

(3224)

Sap I

(3108)

Bst1107 I

(2995)

Tth111 I

(2969)

T7 promoter primer #69348-3

pET upstream primer #69214-3

T7 promoter

lac operator

rbs

Bgl

II

Xba

I

AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGA

Nde

I

Nhe

I

T7•Tag

His•Tag

Nco

I

TATACCATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGGCCTGGTGCCGCGCGGCAGCCATATGGCTAGCATGACTGGTGGACAGCAA

MetGlySerSerHisHisHisHisHisHisSerSerGlyLeuValProArgGlySerHisMetAlaSerMetThrGlyGlyGlnGln

thrombin

Eag

I

His•Tag

Bam

H I

Eco

R I

Sac

I

Not

I

Xho

I

Sal

I

Hin

d III

ATGGGTCGCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCC pET-28a(+)

MetGlyArgGlySerGluPheGluLeuArgArgGlnAlaCysGlyArgThrArgAlaProProProProProLeuArgSerGlyCysEnd

...GGTCGGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCC pET-28b(+)

...GlyArgAspProAsnSerSerSerValAspLysLeuAlaAlaAlaLeuGluHisHisHisHisHisHisEnd

...GGTCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCC pET-28c(+)

...

T7 terminator

Bpu

1102 I

GAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG

T7 terminator primer #69337-3

pET-28a-c(+) cloning/expression region

Novagen

ORDERING

800-526-7319

TECHNICALSUPPORT

800-207-0144

pET-28a(+) Restriction Sites

Enzyme

AccI

AceIII

# SitesLocations

21802994

78901618

31764967

AciI77

AflIII211233224

AluI22

AlwI13

Alw21I7159190

30423542

Alw44I311033038

AlwNI13640

ApaI11334

ApaBI1807

ApoI61921398

4940

AvaI21584298

AvaII516752051

BamHI1198

BanI9253445

17621892

BanII6190507

5202

BbsI412691608

BbvI27

BccI14

Bce83I6211937

3854

BcefI6642983

5153

BcgI9160194

19491983

BclI11137

BfaI770232

40265278

BglI12187

BglII1401

BmgI11332

BpmI49611450

Bpu10I223304443

Bpu1102I180

BsaAI229765127

BsaBI3400406

BsaHI5446467

BsaJI1057296

21963384

BsaWI721442

35774561

BsaXI217825075

BsbI229405034

BscGI11

BsgI39741174

BsiI13397

BsiEI51691908

BslI23

BsmI243104387

BsmAI68201225

4442

BsmBI317382865

BsmFI45842125

BsoFI48

Bsp24I12

Bsp1286I12

BspEI222413

BspGI12750

BspLU11I13224

BsrI22

BsrBI43563157

BsrDI211701536

BsrFI7433442

43805228

BssHII11534

Bst1107I12995

4

Enzyme

BstEII

BstXI

BstYI

Cac8I

CjeI

CjePI

ClaI

CviJI

CviRI

DdeI

DpnI

DraIII

DrdI

DrdII

DsaI

EaeI

EagI

EarI

EciI

Eco47III

Eco57I

EcoNI

EcoO109I

EcoRI

EcoRII

EcoRV

FauI

FokI

FspI

GdiII

HaeI

HaeII

HaeIII

HgaI

HgiEII

HhaI

Hin4I

HincII

HindIII

HinfI

HpaI

HphI

MaeII

MaeIII

MboII

MluI

MmeI

MnlI

MseI

MslI

MspI

MspA1I

5342

MwoI

NarI

NciI

NcoI

NdeI

NgoAIV

NheI

NlaIII

NlaIV

NotI

NruI

NsiI

NspI

39

4

12

1

1

4

1

26

22

1

1

2

4

# Sites

1

3

9

40

26

30

1

86

22

11

21

1

3

2

3

4

1

3

3

3

1

2

3

1

10

1

17

9

1

4

6

14

24

11

2

47

3

2

1

18

1

16

14

16

12

1

7

25

25

6

29

9

Locations

1304

9251054

132198

24163865

1177

401

3876

687

4675

1899

Enzyme

Pfl1108I

PflMI

PleI

PshAI

Psp5II

Psp1406I

PvuI

PvuII

RcaI

RsaI

SacI

SalI

SapI

Sau96I

Sau3AI

ScrFI

SfaNI

SfcI

SgfI

SgrAI

SmaI

SphI

SspI

StyI

TaqI

TaqII

TfiI

1169

2769

2205

166

851

4513

1178

2910

431

2172

2443

4064

563

3239

2505

4670

1797

3250

2583

ThaI

TseI

Tsp45I

3702

Tsp509I

Tth111I

Tth111II

3810

4112

1629

4654

UbaJI

VspI

XbaI

XcmI

XhoI

XmnI

20

1

8

21

5

1

3

1

2

38

27

7

# Sites

1

2

9

1

1

4

1

3

3

3

1

1

1

14

21

22

23

4

1

1

1

1

2

2

15

6

9

Locations

2010

7054689

384672

36034658

1968

2230

7852153

4426

17231816

5213944

12703030

190

179

3108

759

5062

1555

5070

TB074 12/98

3118

2549

2815

4819

4261

4912

623

3538

11072218

4117

4

2139

466

2018

521

1982

2230

580

5164

1334

2342

2509

1043

4081

2107

1610

228

2801

336

3315

3726

1415

2835

2238

3613

4745

1449

5127

2917

846

296

166

166

741

900

528

3772

658

53

192

256

3250

1573

3332

5132

560

431

3108

3298

2029

4338

556

846

3371

5082

2196

563

4239

3444

2478

1797

2230

1161

3384

1701

4314

1758

4671

369

4426

442

4300

598

4351

57

1031

5031

1802

4337

348936805346

4919

296

1249

2104

4393

1922

2274

4565

3126

2778

4656

4680

3199

3719

1304

4573

2969

962

3853

384

335

979

158

2782

2132

5300

266328762971

20842751

1655

4262

1808

1495

4815

2685

4389

1867

1513

38143821

721

1022

181

173

1629

46254814

2421

581

560

4297

1945

1080

566

4298

2413

1763

1758

4699

3430

2384

3

1123

3439

4633

3623

5104

4

Enzymes that do not cut pET28a(+):

AatIIAflIIAgeI

BaeIBsaIBseRI

Bsu36IDraIEam1105I

MscIMunINspV

PmlIPstIRleAI

ScaISexAISfiI

SrfISse8387IStuI

AscI

BspMI

FseI

PacI

RsrII

SnaBI

SunI

AvrII

BsrGI

KpnI

PmeI

SacII

SpeI

SwaI

1351

4442

2495

17382865

1175

2797

84

2815

446

296

238

433

231

12406

264

2934

467

1153

3566

581

1723

3811

1763

1816

2

4825

809

5271

20212181

166

4083

4276

598

4542

256928613228

Novagen

FAX

608-238-1388

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MAIL

novatech@

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