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
-
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
•
E
-
novatech@