pET-3d

Bacterial expression vector with a BamHI cloning site. Same reading frame as pET-3c but with an NcoI site at the start codon.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-3d
ApoI - EcoRI (4635) AatII (4564) ZraI (4562) SspI (4446) ScaI (4122) PvuI (4012) PstI (3887) AhdI (3642) AlwNI (3165) AflIII - PciI (2749) BspQI - SapI (2633) BstZ17I (2520) BsaAI (2501) PflFI - Tth111I (2494) PfoI (2391) BsmBI - Esp3I (2390) BspDI - ClaI (24) HindIII (29) BlpI (458) BamHI (510) NcoI (546) RBS XbaI (585) T7 promoter BglII (643) SgrAI (684) SphI (840) EcoNI (900) SalI (925) PshAI (990) EagI (1213) NruI (1248) BstAPI (1325) BfuAI - BspMI (1328) BsmI (1633) AvaI - BsoBI (1699) MscI * (1720) Bpu10I (1855) BsgI * (1909) PvuII (2340) pET-3d 4637 bp
ApoI  (4635)
1 site		 R A A T T Y Y T T A A R
ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (4635)
1 site		 G A A T T C C T T A A G
AatII  (4564)
1 site		 G A C G T C C T G C A G
ZraI  (4562)
1 site		 G A C G T C C T G C A G
SspI  (4446)
1 site		 A A T A T T T T A T A A
ScaI  (4122)
1 site		 A G T A C T T C A T G A
PvuI  (4012)
1 site		 C G A T C G G C T A G C
PstI  (3887)
1 site		 C T G C A G G A C G T C
AhdI  (3642)
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.
AlwNI  (3165)
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.
AflIII  (2749)
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  (2749)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2633)
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  (2633)
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.
BstZ17I  (2520)
1 site		 G T A T A C C A T A T G
BsaAI  (2501)
1 site		 Y A C G T R R T G C A Y
PflFI  (2494)
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  (2494)
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.
PfoI  (2391)
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.
BsmBI  (2390)
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  (2390)
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.
BspDI  (24)
1 site		 A T C G A T T A G C T A
ClaI  (24)
1 site		 A T C G A T T A G C T A
HindIII  (29)
1 site		 A A G C T T T T C G A A
BlpI  (458)
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.
BamHI  (510)
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.
NcoI  (546)
1 site		 C C A T G G G G T A C C
XbaI  (585)
1 site		 T C T A G A A G A T C T
BglII  (643)
1 site		 A G A T C T T C T A G A
SgrAI  (684)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SphI  (840)
1 site		 G C A T G C C G T A C G
EcoNI  (900)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SalI  (925)
1 site		 G T C G A C C A G C T G
PshAI  (990)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
EagI  (1213)
1 site		 C G G C C G G C C G G C
NruI  (1248)
1 site		 T C G C G A A G C G C T
BstAPI  (1325)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
BfuAI  (1328)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1328)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BsmI  (1633)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
AvaI  (1699)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1699)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
MscI  (1720)
1 site		 T G G C C A A C C G G T
* Blocked by Dcm methylation.
Bpu10I  (1855)
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.
BsgI  (1909)
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).
PvuII  (2340)
1 site		 C A G C T G G T C G A C
AmpR
3569 .. 4429  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   3569 .. 4360  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3569 .. 4429  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   4361 .. 4429  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3569 .. 4429  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2810 .. 3398  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2810 .. 3398  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
2189 .. 2380  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2189 .. 2380  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
4430 .. 4534  =  105 bp
AmpR promoter
4430 .. 4534  =  105 bp
T7 terminator
404 .. 451  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
404 .. 451  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 tag
517 .. 549  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 tag
517 .. 549  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 promoter
610 .. 628  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
610 .. 628  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
557 .. 562  =  6 bp
ribosome binding site
RBS
557 .. 562  =  6 bp
ribosome binding site
ORF:  86 .. 451  =  366 bp
ORF:  121 amino acids  =  12.9 kDa
ORF:  1454 .. 1792  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  2156 .. 2380  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  765 .. 1550  =  786 bp
ORF:  261 amino acids  =  27.5 kDa
ORF:  3699 .. 3965  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  570 .. 839  =  270 bp
ORF:  89 amino acids  =  9.3 kDa
ORF:  897 .. 1355  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  713 .. 1063  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
ORF:  3569 .. 4429  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1789 .. 2157  =  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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