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pGEM®-7Zf(+)

Cloning vector for in vitro RNA synthesis, ssDNA production, and generation of unidirectional deletions. Identical to pGEM®‑7Zf(-) except for the orientation of the f1 origin.

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pGEM-7Zf(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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PspOMI (10) NaeI (2689) NgoMIV (2687) BtgZI (2587) BsaAI - DraIII (2586) PsiI (2458) XmnI (1991) ScaI (1872) TatI (1870) NmeAIII (1540) ApaI (14) BseRI - ZraI (18) AatII (20) SphI (26) XbaI (31) PaeR7I - PspXI - XhoI (37) EcoRI (43) Acc65I (49) KpnI (53) TspMI - XmaI (54) SmaI (56) BstBI (61) BspDI - ClaI (67) HindIII (72) BamHI (78) BspEI * (81) Eco53kI (89) SacI (91) MluI (96) BstXI (100) NsiI (109) BspQI - SapI (383) PciI (499) BseYI (803) PspFI (807) AlwNI (915) AhdI (1392) BsaI (1453) BpmI (1462) pGEM®-7Zf(+) 2997 bp
PspOMI  (10)
1 site
G G G C C C C C C G G G
NaeI  (2689)
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  (2687)
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.
BtgZI  (2587)
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.
BsaAI  (2586)
1 site
Y A C G T R R T G C A Y
DraIII  (2586)
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  (2458)
1 site
T T A T A A A A T A T T
XmnI  (1991)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1872)
1 site
A G T A C T T C A T G A
TatI  (1870)
1 site
W G T A C W W C A T G W
NmeAIII  (1540)
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).
ApaI  (14)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BseRI  (18)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
ZraI  (18)
1 site
G A C G T C C T G C A G
AatII  (20)
1 site
G A C G T C C T G C A G
SphI  (26)
1 site
G C A T G C C G T A C G
XbaI  (31)
1 site
T C T A G A A G A T C T
PaeR7I  (37)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (37)
1 site
V C T C G A G B B G A G C T C V
XhoI  (37)
1 site
C T C G A G G A G C T C
EcoRI  (43)
1 site
G A A T T C C T T A A G
Acc65I  (49)
1 site
G G T A C C C C A T G G
KpnI  (53)
1 site
G G T A C C C C A T G G
TspMI  (54)
1 site
C C C G G G G G G C C C
XmaI  (54)
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.
SmaI  (56)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BstBI  (61)
1 site
T T C G A A A A G C T T
BspDI  (67)
1 site
A T C G A T T A G C T A
ClaI  (67)
1 site
A T C G A T T A G C T A
HindIII  (72)
1 site
A A G C T T T T C G A A
BamHI  (78)
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.
BspEI  (81)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
Eco53kI  (89)
1 site
G A G C T C C T C G A G
SacI  (91)
1 site
G A G C T C C T C G A G
MluI  (96)
1 site
A C G C G T T G C G C A
BstXI  (100)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
NsiI  (109)
1 site
A T G C A T T A C G T A
BspQI  (383)
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  (383)
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.
PciI  (499)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (803)
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  (807)
1 site
C C C A G C G G G T C G
AlwNI  (915)
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  (1392)
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  (1453)
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  (1462)
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
10 .. 110  =  101 bp
multiple cloning site
MCS
10 .. 110  =  101 bp
multiple cloning site
AmpR
1319 .. 2179  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1319 .. 2110  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1319 .. 2179  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2111 .. 2179  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1319 .. 2179  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
560 .. 1148  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
560 .. 1148  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2362 .. 2817  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2362 .. 2817  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
2180 .. 2284  =  105 bp
AmpR promoter
2180 .. 2284  =  105 bp
lac promoter
206 .. 236  =  31 bp
   Segment 3:  -10  
   206 .. 212  =  7 bp
promoter for the E. coli lac operon
lac promoter
206 .. 236  =  31 bp
   Segment 2:  
   213 .. 230  =  18 bp
promoter for the E. coli lac operon
lac promoter
206 .. 236  =  31 bp
   Segment 1:  -35  
   231 .. 236  =  6 bp
promoter for the E. coli lac operon
lac promoter
206 .. 236  =  31 bp
3 segments
promoter for the E. coli lac operon
SP6 promoter
122 .. 140  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
122 .. 140  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
T7 promoter
2981 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2981 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
158 .. 174  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
158 .. 174  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
182 .. 198  =  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
182 .. 198  =  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
2958 .. 2974  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2958 .. 2974  =  17 bp
common sequencing primer, one of multiple similar variants
lacZα
2794 .. 162  =  366 bp
121 amino acids  =  13.7 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2794 .. 162  =  366 bp
121 amino acids  =  13.7 kDa
Product: LacZα fragment of β-galactosidase
ORF:  1449 .. 1715  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2794 .. 162  =  366 bp
ORF:  121 amino acids  =  13.7 kDa
ORF:  2829 .. 110  =  279 bp
ORF:  92 amino acids  =  9.9 kDa
ORF:  2076 .. 2357  =  282 bp
ORF:  93 amino acids  =  10.9 kDa
ORF:  1319 .. 2179  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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