pGEM-11Zf(+)

Vector for standard cloning, efficient in vitro RNA synthesis, and ssDNA production. Identical to pGEM®‑11Zf(-) except for the orientation of the f1 origin.

Sequence Author: Promega

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T7 promoter M13 fwd NaeI (2913) NgoMIV (2911) BtgZI (2811) BsaAI - DraIII (2810) PsiI (2682) BstAPI (2534) NdeI (2533) PfoI (2395) AatII (2284) ZraI (2282) XmnI (1961) ScaI (1842) MscI (11) SfiI (17) Eco53kI (25) SacI (27) EcoRI (29) SalI (35) AccI (36) HincII (37) AbsI - AvaI - BsoBI - PaeR7I - PspXI - XhoI (41) BamHI (47) PspOMI (53) ApaI (57) XbaI (59) NotI (67) SphI (76) NsiI (78) HindIII (80) SP6 promoter lac operator BspQI - SapI (353) AflIII - PciI (469) BseYI (773) PspFI (777) AlwNI (885) AhdI (1362) BsaI (1423) BpmI (1432) pGEM®-11Zf(+) 3221 bp
NaeI  (2913)
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  (2911)
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  (2811)
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  (2810)
1 site
Y A C G T R R T G C A Y
DraIII  (2810)
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  (2682)
1 site
T T A T A A A A T A T T
BstAPI  (2534)
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.
NdeI  (2533)
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.
PfoI  (2395)
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.
AatII  (2284)
1 site
G A C G T C C T G C A G
ZraI  (2282)
1 site
G A C G T C C T G C A G
XmnI  (1961)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1842)
1 site
A G T A C T T C A T G A
MscI  (11)
1 site
T G G C C A A C C G G T
SfiI  (17)
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.
Eco53kI  (25)
1 site
G A G C T C C T C G A G
SacI  (27)
1 site
G A G C T C C T C G A G
EcoRI  (29)
1 site
G A A T T C C T T A A G
SalI  (35)
1 site
G T C G A C C A G C T G
AccI  (36)
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  (37)
1 site
G T Y R A C C A R Y T G
AbsI  (41)
1 site
C C T C G A G G G G A G C T C C
AvaI  (41)
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  (41)
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.
PaeR7I  (41)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (41)
1 site
V C T C G A G B B G A G C T C V
XhoI  (41)
1 site
C T C G A G G A G C T C
BamHI  (47)
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.
PspOMI  (53)
1 site
G G G C C C C C C G G G
ApaI  (57)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
XbaI  (59)
1 site
T C T A G A A G A T C T
NotI  (67)
1 site
G C G G C C G C C G C C G G C G
SphI  (76)
1 site
G C A T G C C G T A C G
NsiI  (78)
1 site
A T G C A T T A C G T A
HindIII  (80)
1 site
A A G C T T T T C G A A
BspQI  (353)
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  (353)
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  (469)
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  (469)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (773)
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  (777)
1 site
C C C A G C G G G T C G
AlwNI  (885)
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  (1362)
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  (1423)
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  (1432)
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
9 .. 85  =  77 bp
multiple cloning site
MCS
9 .. 85  =  77 bp
multiple cloning site
AmpR
1289 .. 2149  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1289 .. 2080  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1289 .. 2149  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2081 .. 2149  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1289 .. 2149  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
530 .. 1118  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
530 .. 1118  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2586 .. 3041  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2586 .. 3041  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
2150 .. 2254  =  105 bp
AmpR promoter
2150 .. 2254  =  105 bp
lac promoter
176 .. 206  =  31 bp
3 segments
   Segment 3:  -10  
   176 .. 182  =  7 bp
promoter for the E. coli lac operon
lac promoter
176 .. 206  =  31 bp
3 segments
   Segment 2:  
   183 .. 200  =  18 bp
promoter for the E. coli lac operon
lac promoter
176 .. 206  =  31 bp
3 segments
   Segment 1:  -35  
   201 .. 206  =  6 bp
promoter for the E. coli lac operon
lac promoter
176 .. 206  =  31 bp
3 segments
promoter for the E. coli lac operon
SP6 promoter
92 .. 110  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
92 .. 110  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
T7 promoter
3205 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3205 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
128 .. 144  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
128 .. 144  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
152 .. 168  =  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
152 .. 168  =  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
3182 .. 3198  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
3182 .. 3198  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1419 .. 1685  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1289 .. 2149  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3018 .. 132  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
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