Resources
Plasmid Files

pGEM®-9Zf(-)

Cloning vector with a compact MCS for in vitro RNA synthesis, ssDNA production, and generation of unidirectional deletions.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pGEM-9Zf(-) Sequence and MappGEM-9Zf(-).dna
Map and Sequence File   
Sequence Author:  Promega
Download Free Trial Get SnapGene Viewer

 EagI - NotI (2887) PflFI - Tth111I (2879) PsiI (2613) DraIII (2488) BsaAI (2485) BtgZI (2480) NaeI (2382) NgoMIV (2380) XmnI (1935) BsaHI (1873) ScaI (1816) TatI (1814) NmeAIII (1484) NsiI (9) SpeI (12) HindIII (18) XbaI (27) EcoRI (36) SalI - SgrDI (42) AccI (43) HincII (44) Eco53kI (50) SacI (52) SfiI (82) MscI (85) M13 rev lac operator BspQI - SapI (327) AflIII - PciI (443) NspI (447) BseYI (747) PspFI (751) AlwNI (859) AhdI (1336) BsaI (1397) BpmI (1406) pGEM®-9Zf(-) 2912 bp
EagI  (2887)
1 site
C G G C C G G C C G G C
NotI  (2887)
1 site
G C G G C C G C C G C C G G C G
PflFI  (2879)
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  (2879)
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.
PsiI  (2613)
1 site
T T A T A A A A T A T T
DraIII  (2488)
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.
BsaAI  (2485)
1 site
Y A C G T R R T G C A Y
BtgZI  (2480)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
NaeI  (2382)
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  (2380)
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.
XmnI  (1935)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (1873)
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  (1816)
1 site
A G T A C T T C A T G A
TatI  (1814)
1 site
W G T A C W W C A T G W
NmeAIII  (1484)
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).
NsiI  (9)
1 site
A T G C A T T A C G T A
SpeI  (12)
1 site
A C T A G T T G A T C A
HindIII  (18)
1 site
A A G C T T T T C G A A
XbaI  (27)
1 site
T C T A G A A G A T C T
EcoRI  (36)
1 site
G A A T T C C T T A A G
SalI  (42)
1 site
G T C G A C C A G C T G
SgrDI  (42)
1 site
C G T C G A C G G C A G C T G C
AccI  (43)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HincII  (44)
1 site
G T Y R A C C A R Y T G
Eco53kI  (50)
1 site
G A G C T C C T C G A G
SacI  (52)
1 site
G A G C T C C T C G A G
SfiI  (82)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
MscI  (85)
1 site
T G G C C A A C C G G T
BspQI  (327)
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  (327)
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  (443)
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  (443)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (447)
1 site
R C A T G Y Y G T A C R
BseYI  (747)
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  (751)
1 site
C C C A G C G G G T C G
AlwNI  (859)
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.
AhdI  (1336)
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.
BsaI  (1397)
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.
BpmI  (1406)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
MCS
5 .. 53  =  49 bp
multiple cloning site
MCS
5 .. 53  =  49 bp
multiple cloning site
AmpR
1263 .. 2123  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1263 .. 2054  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1263 .. 2123  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2055 .. 2123  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1263 .. 2123  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
504 .. 1092  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
504 .. 1092  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2255 .. 2710  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2255 .. 2710  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
2124 .. 2228  =  105 bp
AmpR promoter
2124 .. 2228  =  105 bp
lac promoter
150 .. 180  =  31 bp
   Segment 3:  -10  
   150 .. 156  =  7 bp
promoter for the E. coli lac operon
lac promoter
150 .. 180  =  31 bp
   Segment 2:  
   157 .. 174  =  18 bp
promoter for the E. coli lac operon
lac promoter
150 .. 180  =  31 bp
   Segment 1:  -35  
   175 .. 180  =  6 bp
promoter for the E. coli lac operon
lac promoter
150 .. 180  =  31 bp
3 segments
promoter for the E. coli lac operon
T7 promoter
54 .. 72  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
54 .. 72  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
2896 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
2896 .. 2  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 rev
102 .. 118  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
102 .. 118  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
126 .. 142  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
126 .. 142  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
M13 fwd
2851 .. 2867  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
2851 .. 2867  =  17 bp
common sequencing primer, one of multiple similar
variants
lacZα
2702 .. 101  =  312 bp
103 amino acids  =  11.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2702 .. 101  =  312 bp
103 amino acids  =  11.6 kDa
Product: LacZα fragment of β-galactosidase
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file 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.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter