TB051 12/98
The pET-23a-d(+) vectors carry an N-terminal T7Tag sequence plus an optional C-terminal
HisTag sequence. These vectors differ from pET-21a-d(+) by the plain T7 promoter instead
of the T7lac promoter and by the absence of the lacI gene. 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. 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).
Cat. No.
69745-3
69746-3
69747-3
69748-3
pET-23a DNA
pET-23b DNA
pET-23c DNA
pET-23d DNA
303-319
302
207-239
158-203
140-157
26-72
1450
2211-3068
3200-3655
Sty I(57)
Bpu1102 I(80)
NgoA IV(3525)
Drd I(3429)
Dra III(3424)
f1
5)
0-365
320
(
n
igi
or
BsaH I(2818)
Sca I(2761)
Pvu II(979)
1 130
Pst I(2526)
(3666bp)
Tth111 I(1133)
Bst1107 I(1159)
45
68
0)
Pvu I(2651)
pET-23a(+)
Ap (22
Xho I(158)
Not I(166)
Eag I(166)
Hind III(173)
Sal I(179)
Hinc II(181)
Sac I(190)
EcoR I(192)
BamH I(198)
Nhe I(231)
Nde I(238)
Xba I(276)
Bgl II(334)
Msc I(359)
Dsa I(360)
Bpu10 I(494)
Bbs I(506)
Bsg I(548)
Eco47 III(642)
(1
T7 promoter
T7 transcription start
T7Tag coding sequence
Multiple cloning sites
(BamH I - Xho I)
HisTag coding sequence
T7 terminator
pBR322 origin
bla coding sequence
f1 origin
Bgl I(2401)
or
Sap I(1272)
Afl III(1388)
BspLU11 I(1388)
Ahd I(2281)
AlwN I(1804)
Bgl II
rbs
Xba I
AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGAGACCACAACGGTTTCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGA
Nde I Nhe I
pET-23a
T7Tag
Hinc II
Sal I Hind III
Eag I
Not I
Xho I
HisTag
TATACATATGGCTAGCATGACTGGTGGACAGCAAATGGGTCGCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGA
MetAlaSerMetThrGlyGlyGlnGlnMetGlyArgGlySerGluPheGluLeuArgArgGlnAlaCysGlyArgThrArgAlaProProProProProLeu
pET-23d
Nco I
...TACCATGGCTAGC...
MetAlaSer...
pET-23b
...GGTCGGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGA
...GlyArgAspProAsnSerSerSerValAspLysLeuAlaAlaAlaLeuGluHisHisHisHisHisHisEnd
pET-23c,d
...GGTCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGA
...GlyArgIleArgIleArgAlaProSerThrSerLeuArgProHisSerSerThrThrThrThrThrThrGlu
T7 terminator
Bpu1102 I
GATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG
T7 terminator primer #69337-3
TECHNICAL SUPPORT
800-207-0144
# Sites
2
5
49
1
18
13
7
Alw44I
AlwNI
ApoI
AvaI
AvaII
BamHI
BanI
BanII
BbsI
BbvI
BccI
Bce83I
BcefI
BcgI
3
1
3
2
5
1
2
2
1
21
5
5
2
7
BfaI
BglI
BglII
BpmI
Bpu10I
Bpu1102I
BsaI
BsaAI
BsaBI
BsaHI
BsaJI
BsaWI
BsaXI
BsbI
BscGI
1
1
2
1
1
2
2
2
1
3
5
1
2
10
BsgI
BsiI
BsiEI
BslI
BsmAI
BsmBI
BsmFI
BsoFI
Bsp24I
1
2
5
12
4
1
2
33
6
Bsp1286I 8
BspEI
BspGI
BspLU11I
BsrI
BsrBI
BsrDI
BsrFI
Bst1107I
BstYI
2
1
1
15
3
2
2
1
10
Cac8I
CjeI
CjePI
CviJI
CviRI
DdeI
16
12
12
56
13
10
DpnI
DraI
DraIII
19
3
1
Locations
180
1158
897
1038
1340
2580
3264
1388
159
2867
1202
1804
192
158
352
198
2229
190
506
2318
21
1890
160
2786
70
2136
2401
334
915
494
80
301
1140
333
2818
57
2
3372
1104
44
2061
548
1561
169
190
2952
1702
382
3226
338
394
3237
1206
1706
2948
673
2419
2641
3461
3499
2442
1479
3450
194
2820
232
2471
2729
1777
228
277
3575
3414
2018
965
402
3431
2886
1883
2342
3424
585
3331
364
2282
1548
1594
749
2306
1741
1082
2828
Enzyme
DrdI
DrdII
DsaI
EaeI
EagI
Eam1105I
EarI
EciI
Eco47III
Eco57I
EcoO109I
EcoRI
EcoRII
FauI
# Sites
3
1
1
3
1
1
2
3
1
2
3
1
4
8
FokI
FspI
GdiII
HaeI
HaeII
2
2
4
6
HaeIII
HgaI
13
6
HgiEII
HhaI
Hin4I
HincII
HindIII
HinfI
1
23
2
1
1
9
HphI
MaeII
MaeIII
MboII
12
15
8
MmeI
MnlI
MscI
MseI
MslI
3
19
1
22
6
MspI
MspA1I
17
6
MwoI
NciI
18
7
NdeI
NgoAIV
NheI
NlaIII
NlaIV
NotI
NspI
Pfl1108I
PleI
1
1
1
15
15
1
3
1
6
Psp5II
Psp1406I
PstI
PvuI
PvuII
RcaI
RsaI
SacI
SalI
SapI
Sau96I
2
4
1
1
1
2
2
1
1
1
10
999
2351
360
577
TB051 12/98
2572
1715
3518
2945
1304
1728
2651
301
1029
659
1029
2342
3118
201
2091
159
2867
2
914
1388
233
1881
1913
2059
190
2952
577
382
3499
1206
1706
1321
2342
2361
1159
132
2040
3122
2516
3525
3568
198
2126
334
2138
2800
3639
80
1196
101
1663
136
2072
2147
3424
2166
2858
580
2906
494
2238
2029
2923
656
2778
Locations
1081 1496
3429
360
166
357
166
2281
1272 3076
1462 1608
642
1936 2948
53
352
192
354
1414
318
403
1101 3561
607
669
2247 2428
369
2503
166
2669
359
1403
561
644
3583
3379
2669
2436
Enzyme
Sau3AI
ScaI
ScrFI
SfaNI
SfcI
SspI
StyI
TaqI
# Sites
19
1
11
13
5
2
1
7
TaqII
TfiI
ThaI
TseI
Tsp45I
3
17
21
6
Tsp509I
Tth111I
Tth111II
UbaJI
VspI
XbaI
XhoI
XmnI
1
4
12
2
1
1
2
394
1535
684
3630
747
2715
1548
870
1091
933
1074
1414
1266
1866
1636
1499
2077
3575
924
3641
1974
1081
2807
2280
181
173
309
1759
433
2767
2354
438
2276
1008
2973
942
3351
1017
3008
1288
3373
2124
3425
1363
506
2954
1603
1259
3063
1787
2050
3564
3401
2121
2876
375
3051
570
961
2533
2692
84
2916
979
1098
1730
1975
398
2464
238
3525
231
726
2815
1032
1067
1768
1025
1392
1282
1767
2270
3359
394
2507
2880
3209
394
2419
673
2641
2351
3116
2761
352
2402
2761
302
3085
57
159
2932
1290
3328
438
1653
3216
1844
2522
3643
180
3457
2629
190
331
1488
2814
2967
2984
942
1363
827
3597
192
2709
1133
849
1040
1135
2537
2748
268
3200
318
3226
2148
3237
2454
1978
1985
2017
317
276
158
946
2453
2880
359
166
733
2299
317
3367
352
713
2526
2651
979
2108
1194
190
179
1272
53
2323
Locations
860
3415
ApaBI
BmgI
BssHII
EcoNI
MluI
NsiI
PmlI
SexAI
SnaBI
StuI