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Plasmid Files

pET-3c

Bacterial expression vector with a BamHI cloning site. For alternative reading frames, use pET‑3a or pET‑3b.

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pET-3c Sequence and MappET-3c.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 ApoI - EcoRI (4636) AatII (4565) ZraI (4563) SspI (4447) ScaI (4123) PvuI (4013) PstI (3888) AhdI (3643) AlwNI (3166) AflIII - PciI (2750) BspQI - SapI (2634) BstZ17I (2521) BsaAI (2502) PflFI - Tth111I (2495) PfoI (2392) BsmBI (2391) BspDI - ClaI (24) HindIII (29) BlpI (458) BamHI (510) NdeI (548) RBS XbaI (586) T7 promoter BglII (644) SgrAI (685) SphI (841) EcoNI (901) SalI (926) PshAI (991) EagI (1214) NruI (1249) BstAPI (1326) BfuAI - BspMI (1329) BsmI (1634) AvaI - BsoBI (1700) BmeT110I (1701) MscI * (1721) Bpu10I (1856) BsgI * (1910) PvuII (2341) pET-3c 4638 bp
ApoI  (4636)
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  (4636)
1 site
G A A T T C C T T A A G
AatII  (4565)
1 site
G A C G T C C T G C A G
ZraI  (4563)
1 site
G A C G T C C T G C A G
SspI  (4447)
1 site
A A T A T T T T A T A A
ScaI  (4123)
1 site
A G T A C T T C A T G A
PvuI  (4013)
1 site
C G A T C G G C T A G C
PstI  (3888)
1 site
C T G C A G G A C G T C
AhdI  (3643)
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  (3166)
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  (2750)
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  (2750)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2634)
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  (2634)
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  (2521)
1 site
G T A T A C C A T A T G
BsaAI  (2502)
1 site
Y A C G T R R T G C A Y
PflFI  (2495)
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  (2495)
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  (2392)
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  (2391)
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.
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.
NdeI  (548)
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  (586)
1 site
T C T A G A A G A T C T
BglII  (644)
1 site
A G A T C T T C T A G A
SgrAI  (685)
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  (841)
1 site
G C A T G C C G T A C G
EcoNI  (901)
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  (926)
1 site
G T C G A C C A G C T G
PshAI  (991)
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  (1214)
1 site
C G G C C G G C C G G C
NruI  (1249)
1 site
T C G C G A A G C G C T
BstAPI  (1326)
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  (1329)
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  (1329)
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  (1634)
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  (1700)
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  (1700)
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.
BmeT110I  (1701)
1 site
C Y C G R G G R G C Y C
MscI  (1721)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
Bpu10I  (1856)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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  (1910)
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  (2341)
1 site
C A G C T G G T C G A C
AmpR
3570 .. 4430  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3570 .. 4361  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3570 .. 4430  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4362 .. 4430  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3570 .. 4430  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2811 .. 3399  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2811 .. 3399  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rop
2190 .. 2381  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2190 .. 2381  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
4431 .. 4535  =  105 bp
AmpR promoter
4431 .. 4535  =  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
611 .. 629  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
611 .. 629  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
558 .. 563  =  6 bp
ribosome binding site
RBS
558 .. 563  =  6 bp
ribosome binding site
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