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Plasmid Files

pET-9a

Bacterial expression vector with a BamHI cloning site. For alternative reading frames, use pET‑9b or pET‑9c.

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pET-9a Sequence and MappET-9a.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 EcoRI (4339) AsiSI - PvuI (3954) SspI (3879) SmaI (3828) TspMI - XmaI (3826) PaeR7I - PspXI - XhoI (3552) AcuI (3300) AlwNI (3168) BciVI (2955) BssS α I (2925) AflIII - PciI (2752) BsrBI (2685) BspQI - SapI (2636) TatI (2556) BstZ17I (2523) BsaAI (2504) PflFI - Tth111I (2497) PfoI (2394) PvuII (2343) BlpI (458) BamHI (510) NdeI (550) RBS XbaI (588) BsaI (613) T7 promoter BglII (646) SgrAI (687) SphI (843) SalI (928) HincII (930) PshAI (993) EagI (1216) BstAPI (1328) BfuAI - BspMI (1331) MscI * (1723) BsgI * (1912) XmnI (2310) pET-9a 4341 bp
EcoRI  (4339)
1 site
G A A T T C C T T A A G
AsiSI  (3954)
1 site
G C G A T C G C C G C T A G C G
PvuI  (3954)
1 site
C G A T C G G C T A G C
SspI  (3879)
1 site
A A T A T T T T A T A A
SmaI  (3828)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3826)
1 site
C C C G G G G G G C C C
XmaI  (3826)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
PaeR7I  (3552)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3552)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3552)
1 site
C T C G A G G A G C T C
AcuI  (3300)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (3168)
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.
BciVI  (2955)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
BssSαI  (2925)
1 site
C A C G A G G T G C T C
AflIII  (2752)
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  (2752)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BsrBI  (2685)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures
up to 50°C.
BspQI  (2636)
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  (2636)
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.
TatI  (2556)
1 site
W G T A C W W C A T G W
BstZ17I  (2523)
1 site
G T A T A C C A T A T G
BsaAI  (2504)
1 site
Y A C G T R R T G C A Y
PflFI  (2497)
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  (2497)
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  (2394)
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.
PvuII  (2343)
1 site
C A G C T G G T C G A C
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  (550)
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  (588)
1 site
T C T A G A A G A T C T
BsaI  (613)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BglII  (646)
1 site
A G A T C T T C T A G A
SgrAI  (687)
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  (843)
1 site
G C A T G C C G T A C G
SalI  (928)
1 site
G T C G A C C A G C T G
HincII  (930)
1 site
G T Y R A C C A R Y T G
PshAI  (993)
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  (1216)
1 site
C G G C C G G C C G G C
BstAPI  (1328)
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  (1331)
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  (1331)
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.
MscI  (1723)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BsgI  (1912)
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).
XmnI  (2310)
1 site
G A A N N N N T T C C T T N N N N A A G
KanR
3523 .. 4338  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
3523 .. 4338  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
2813 .. 3401  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2813 .. 3401  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rop
2192 .. 2383  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2192 .. 2383  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
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
519 .. 551  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 tag
519 .. 551  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 promoter
613 .. 631  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
613 .. 631  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
560 .. 565  =  6 bp
ribosome binding site
RBS
560 .. 565  =  6 bp
ribosome binding site
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