pET-23a(+)

Bacterial vector for expression of N-terminally T7-tagged proteins. For other reading frames, use pET-23b(+) or pET-23c(+).

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-23a(+)
StyI (57) NaeI (3527) NgoMIV (3525) DraIII (3424) PsiI (3296) BsaHI (2818) ScaI (2761) PvuI (2651) PstI (2526) NmeAIII (2429) BglI (2401) AhdI (2281) BlpI (80) 6xHis PaeR7I - PspXI - XhoI (158) EagI - NotI (166) HindIII (173) SalI (179) MCS HincII (181) Eco53kI (188) SacI (190) EcoRI (192) BamHI (198) T7 tag NheI (231) BmtI (235) NdeI (238) RBS XbaI (276) T7 promoter BglII (334) MscI * (359) BtgI (360) FspAI (369) Bpu10I (494) BbsI (506) BsgI * (548) AfeI (642) PvuII (979) BsmBI - Esp3I (1029) PfoI (1030) PflFI - Tth111I (1133) BstZ17I (1159) BspQI - SapI (1272) AflIII - PciI (1388) BseYI (1692) PspFI (1696) AlwNI (1804) pET-23a(+) 3666 bp
StyI  (57)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
NaeI  (3527)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (3525)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
DraIII  (3424)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
PsiI  (3296)
1 site		 T T A T A A A A T A T T
BsaHI  (2818)
1 site		 G R C G Y C C Y G C R G
BsaHI is typically used at 37°C, but is even more active at 60°C.
ScaI  (2761)
1 site		 A G T A C T T C A T G A
PvuI  (2651)
1 site		 C G A T C G G C T A G C
PstI  (2526)
1 site		 C T G C A G G A C G T C
NmeAIII  (2429)
1 site		 G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19
Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BglI  (2401)
1 site		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
AhdI  (2281)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BlpI  (80)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
PaeR7I  (158)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (158)
1 site		 C T C G A G G A G C T C
EagI  (166)
1 site		 C G G C C G G C C G G C
NotI  (166)
1 site		 G C G G C C G C C G C C G G C G
HindIII  (173)
1 site		 A A G C T T T T C G A A
SalI  (179)
1 site		 G T C G A C C A G C T G
HincII  (181)
1 site		 G T Y R A C C A R Y T G
Eco53kI  (188)
1 site		 G A G C T C C T C G A G
SacI  (190)
1 site		 G A G C T C C T C G A G
EcoRI  (192)
1 site		 G A A T T C C T T A A G
BamHI  (198)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
NheI  (231)
1 site		 G C T A G C C G A T C G
BmtI  (235)
1 site		 G C T A G C C G A T C G
NdeI  (238)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
XbaI  (276)
1 site		 T C T A G A A G A T C T
BglII  (334)
1 site		 A G A T C T T C T A G A
MscI  (359)
1 site		 T G G C C A A C C G G T
* Blocked by Dcm methylation.
BtgI  (360)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
FspAI  (369)
1 site		 R T G C G C A Y Y A C G C G T R
Bpu10I  (494)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BbsI  (506)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsgI  (548)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
* Blocked by EcoKI methylation.
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AfeI  (642)
1 site		 A G C G C T T C G C G A
PvuII  (979)
1 site		 C A G C T G G T C G A C
BsmBI  (1029)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (1029)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
PfoI  (1030)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
PflFI  (1133)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1133)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BstZ17I  (1159)
1 site		 G T A T A C C A T A T G
BspQI  (1272)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (1272)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
AflIII  (1388)
1 site		 A C R Y G T T G Y R C A
Sticky ends from different AflIII sites may not be compatible.
PciI  (1388)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BseYI  (1692)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI  (1696)
1 site		 C C C A G C G G G T C G
AlwNI  (1804)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
AmpR
2208 .. 3068  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2208 .. 2999  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2208 .. 3068  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   3000 .. 3068  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2208 .. 3068  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1449 .. 2037  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1449 .. 2037  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3200 .. 3655  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3200 .. 3655  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
828 .. 1019  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
828 .. 1019  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
3069 .. 3173  =  105 bp
AmpR promoter
3069 .. 3173  =  105 bp
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
MCS
158 .. 203  =  46 bp
multiple cloning site
MCS
158 .. 203  =  46 bp
multiple cloning site
T7 tag
207 .. 239  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 tag
207 .. 239  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 promoter
301 .. 319  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
301 .. 319  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
248 .. 253  =  6 bp
ribosome binding site
RBS
248 .. 253  =  6 bp
ribosome binding site
ORF:  2338 .. 2604  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  795 .. 1019  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  2208 .. 3068  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  428 .. 796  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
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Individual Sequences & Maps

pACYCDuet-1
pET-24a(+)
pET-3a
pET-51b(+) Ek_LIC (linearized)
pCDF-1b
pET-24b(+)
pET-3b
pET-52b(+)
pCDF-2 Ek_LIC
pET-24c(+)
pET-3c
pET-52b(+) 3C_LIC
pCDF-2 Ek_LIC (linearized)
pET-24d(+)
pET-3d
pET-52b(+) 3C_LIC (linearized)
pCDFDuet-1
pET-25b(+)
pET-40b(+)
pET-53-DEST
pCOLA-2-DEST
pET-26b(+)
pET-41 Ek_LIC
pET-54-DEST
pCOLA-3-DEST
pET-27b(+)
pET-41 Ek_LIC (linearized)
pET-55-DEST
pCOLADuet-1
pET-28a(+)
pET-41a(+)
pET-56-DEST
pET-11a
pET-28b(+)
pET-41b(+)
pET-57-DEST
pET-11b
pET-28c(+)
pET-41c(+)
pET-58-DEST
pET-11c
pET-29a(+)
pET-42a(+)
pET-59-DEST
pET-11d
pET-29b(+)
pET-42b(+)
pET-60-DEST
pET-14b
pET-29c(+)
pET-42c(+)
pET-61-DEST
pET-15b
pET-30 Ek_LIC
pET-43.1 Ek_LIC
pET-62-DEST
pET-16b
pET-30 Ek_LIC (linearized)
pET-43.1 Ek_LIC (linearized)
pET-9a
pET-17b
pET-30 Xa_LIC
pET-43.1a(+)
pET-9b
pET-19b
pET-30 Xa_LIC (linearized)
pET-43.1b(+)
pET-9c
pET-20b(+)
pET-30a(+)
pET-43.1c(+)
pET-9d
pET-21(+)
pET-30b(+)
pET-44a(+)
pETBlue-1
pET-21a(+)
pET-30c(+)
pET-44b(+)
pETBlue-2
pET-21b(+)
pET-31b(+)
pET-44c(+)
pETcoco-1
pET-21c(+)
pET-32 Ek_LIC
pET-45b(+)
pETcoco-2
pET-21d(+)
pET-32 Ek_LIC (linearized)
pET-46 Ek_LIC
pETDuet-1
pET-22b(+)
pET-32 Xa_LIC
pET-46 Ek_LIC (linearized)
pLacI
pET-23(+)
pET-32 Xa_LIC (linearized)
pET-47b(+)
pLysE
pET-23a(+)
pET-32a(+)
pET-48b(+)
pLysS
pET-23b(+)
pET-32b(+)
pET-49b(+)
pRSF-1b
pET-23c(+)
pET-32c(+)
pET-50b(+)
pRSF-2 Ek_LIC
pET-23d(+)
pET-33b(+)
pET-51b(+)
pRSF-2 Ek_LIC (linearized)
pET-24(+)
pET-39b(+)
pET-51b(+) Ek_LIC
pRSFDuet-1

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