pET-23d(+)

Bacterial vector for expression of N-terminally T7-tagged proteins. Same reading frame as pET-23c(+) but with an NcoI site at the start codon.

Sequence Author: MilliporeSigma (Novagen)

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: pET & Duet Vectors (Novagen) | More Plasmid Sets
No matches
BlpI (80) NaeI (3524) NgoMIV (3522) DraIII (3421) PsiI (3293) BsaHI (2815) ScaI (2758) PvuI (2648) PstI (2523) NmeAIII (2426) BglI (2398) AhdI (2278) 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 (229) BmtI (233) NcoI (234) RBS XbaI (273) T7 promoter BglII (331) MscI * (356) FspAI (366) Bpu10I (491) BbsI (503) BsgI * (545) AfeI (639) PvuII (976) BsmBI - Esp3I (1026) PfoI (1027) PflFI - Tth111I (1130) BstZ17I (1156) BspQI - SapI (1269) AflIII - PciI (1385) BseYI (1689) PspFI (1693) AlwNI (1801) pET-23d(+) 3663 bp
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.
NaeI  (3524)
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  (3522)
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  (3421)
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  (3293)
1 site
T T A T A A A A T A T T
BsaHI  (2815)
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  (2758)
1 site
A G T A C T T C A T G A
PvuI  (2648)
1 site
C G A T C G G C T A G C
PstI  (2523)
1 site
C T G C A G G A C G T C
NmeAIII  (2426)
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  (2398)
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  (2278)
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.
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  (229)
1 site
G C T A G C C G A T C G
BmtI  (233)
1 site
G C T A G C C G A T C G
NcoI  (234)
1 site
C C A T G G G G T A C C
XbaI  (273)
1 site
T C T A G A A G A T C T
BglII  (331)
1 site
A G A T C T T C T A G A
MscI  (356)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
FspAI  (366)
1 site
R T G C G C A Y Y A C G C G T R
Bpu10I  (491)
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  (503)
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  (545)
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  (639)
1 site
A G C G C T T C G C G A
PvuII  (976)
1 site
C A G C T G G T C G A C
BsmBI  (1026)
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  (1026)
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  (1027)
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  (1130)
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  (1130)
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  (1156)
1 site
G T A T A C C A T A T G
BspQI  (1269)
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  (1269)
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  (1385)
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  (1385)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1689)
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  (1693)
1 site
C C C A G C G G G T C G
AlwNI  (1801)
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
2205 .. 3065  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2205 .. 2996  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2205 .. 3065  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   2997 .. 3065  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2205 .. 3065  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1446 .. 2034  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1446 .. 2034  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3197 .. 3652  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3197 .. 3652  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
825 .. 1016  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
825 .. 1016  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
3066 .. 3170  =  105 bp
AmpR promoter
3066 .. 3170  =  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
205 .. 237  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 tag
205 .. 237  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 promoter
298 .. 316  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
298 .. 316  =  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
245 .. 250  =  6 bp
ribosome binding site
RBS
245 .. 250  =  6 bp
ribosome binding site
ORF:  2335 .. 2601  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  792 .. 1016  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  2205 .. 3065  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  425 .. 793  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
Click here to try SnapGene

Download pET-23d(+).dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

The maps, notes, and annotations in the zip file on this page are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as ’’www.snapgene.com/resources’’. Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Discover the most user-friendly molecular biology experience.