pDrive

Bacterial vector for UA cloning and expression of a PCR product.

Sequence Author: Qiagen

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lac operator BspQI - SapI (3842) PciI (3725) PspFI (3425) BseYI (3421) AlwNI (3316) PflMI (2875) Bpu10I (2629) BsmBI (2628) AsiSI (2612) EcoNI (2524) SmaI (2486) TspMI - XmaI (2484) BspDI - ClaI (2303) NruI (2269) AhdI (1962) T7 promoter Acc65I (266) KpnI (270) SphI (278) PstI (283) MluI (285) SnaBI (293) BamHI (299) EcoRI (306) EcoRV (316) EcoRI (321) SalI (327) AccI (328) HincII (329) HindIII (333) PaeR7I - XhoI (339) AvrII - StyI (345) NheI (350) BmtI (354) XbaI (356) AleI - PmlI (367) BstXI (369) EcoO109I - PspOMI (374) ApaI (378) Eco53kI (382) SacI (384) EagI - NotI (387) SP6 promoter AanI - PsiI (684) DraIII (812) BtgZI (813) NgoMIV (913) NaeI (915) BsaHI (1422) TatI (1479) ScaI (1481) NmeAIII (1815) BpmI (1893) BsaI (1896) pDrive 3851 bp
BspQI  (3842)
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  (3842)
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  (3725)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (3425)
1 site
C C C A G C G G G T C G
BseYI  (3421)
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.
AlwNI  (3316)
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.
PflMI  (2875)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
Bpu10I  (2629)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
BsmBI  (2628)
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.
BsmBI-v2 is an improved version of BsmBI.
AsiSI  (2612)
1 site
G C G A T C G C C G C T A G C G
EcoNI  (2524)
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.
SmaI  (2486)
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  (2484)
1 site
C C C G G G G G G C C C
XmaI  (2484)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
BspDI  (2303)
1 site
A T C G A T T A G C T A
ClaI  (2303)
1 site
A T C G A T T A G C T A
NruI  (2269)
1 site
T C G C G A A G C G C T
AhdI  (1962)
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.
Acc65I  (266)
1 site
G G T A C C C C A T G G
KpnI  (270)
1 site
G G T A C C C C A T G G
SphI  (278)
1 site
G C A T G C C G T A C G
PstI  (283)
1 site
C T G C A G G A C G T C
MluI  (285)
1 site
A C G C G T T G C G C A
SnaBI  (293)
1 site
T A C G T A A T G C A T
BamHI  (299)
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.
EcoRI  (306)
2 sites
G A A T T C C T T A A G
EcoRV  (316)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
EcoRI  (321)
2 sites
G A A T T C C T T A A G
SalI  (327)
1 site
G T C G A C C A G C T G
AccI  (328)
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  (329)
1 site
G T Y R A C C A R Y T G
HindIII  (333)
1 site
A A G C T T T T C G A A
PaeR7I  (339)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (339)
1 site
C T C G A G G A G C T C
AvrII  (345)
1 site
C C T A G G G G A T C C
StyI  (345)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NheI  (350)
1 site
G C T A G C C G A T C G
BmtI  (354)
1 site
G C T A G C C G A T C G
XbaI  (356)
1 site
T C T A G A A G A T C T
AleI  (367)
1 site
C A C N N N N G T G G T G N N N N C A C
PmlI  (367)
1 site
C A C G T G G T G C A C
BstXI  (369)
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.
EcoO109I  (374)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (374)
1 site
G G G C C C C C C G G G
ApaI  (378)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
Eco53kI  (382)
1 site
G A G C T C C T C G A G
SacI  (384)
1 site
G A G C T C C T C G A G
EagI  (387)
1 site
C G G C C G G C C G G C
NotI  (387)
1 site
G C G G C C G C C G C C G G C G
AanI  (684)
1 site
T T A T A A A A T A T T
PsiI  (684)
1 site
T T A T A A A A T A T T
DraIII  (812)
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.
BtgZI  (813)
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.
NgoMIV  (913)
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.
NaeI  (915)
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.
BsaHI  (1422)
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.
TatI  (1479)
1 site
W G T A C W W C A T G W
ScaI  (1481)
1 site
A G T A C T T C A T G A
NmeAIII  (1815)
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).
BpmI  (1893)
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.
BsaI  (1896)
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.
AmpR
1175 .. 2035  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1175 .. 1243  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1175 .. 2035  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1244 .. 2035  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1175 .. 2035  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
KanR
2181 .. 2996  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
2181 .. 2996  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
3081 .. 3669  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3081 .. 3669  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
588 .. 1043  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
588 .. 1043  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
MCS
266 .. 393  =  128 bp
multiple cloning site
MCS
266 .. 393  =  128 bp
multiple cloning site
AmpR promoter
1070 .. 1174  =  105 bp
AmpR promoter
1070 .. 1174  =  105 bp
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 1:  -35  
   142 .. 147  =  6 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 2:  
   148 .. 165  =  18 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 3:  -10  
   166 .. 172  =  7 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
promoter for the E. coli lac operon
T7 promoter
239 .. 257  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
239 .. 257  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
399 .. 417  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
399 .. 417  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
lac operator
180 .. 196  =  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
180 .. 196  =  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 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
431 .. 447  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
431 .. 447  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1175 .. 2035  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  216 .. 596  =  381 bp
ORF:  126 amino acids  =  14.5 kDa
ORF:  2181 .. 2996  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  1639 .. 1905  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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