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Plasmid Files

pIEx™-10

Vector for high-level expression in insect cells of secreted proteins with a cleavable N-terminal Strep-Tag® II plus a C-terminal 10xHis tag.

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pIEx-10.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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BmeT110I (3797) AvaI - BsoBI - TspMI - XmaI (3796) BspQI - SapI (3552) PciI (3435) DrdI (3333) AlwNI (3026) AhdI (2547) BsaI (2481) BsrFI (2462) NmeAIII (2400) ScaI (2066) XmnI (1947) SmaI (3798) BspDI * - ClaI * (207) EcoRV (239) PsiI (409) StyI (568) NheI (606) BmtI (610) SphI (614) BsmI (775) BclI * (778) BsiWI (784) MluI (872) BstBI (1172) Acc65I (1200) BsgI (1202) KpnI (1204) BamHI (1206) PshAI (1216) SalI (1219) AccI (1220) HindIII (1225) NotI (1232) Eco53kI (1243) BanII - BseRI - SacI (1245) DraIII (1302) Bsu36I (1312) PacI (1341) PflFI - Tth111I (1565) BtgZI (1588) XbaI (1620) pIEx™-10 3801 bp
BmeT110I  (3797)
1 site
C Y C G R G G R G C Y C
AvaI  (3796)
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  (3796)
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.
TspMI  (3796)
1 site
C C C G G G G G G C C C
XmaI  (3796)
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.
BspQI  (3552)
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  (3552)
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  (3435)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (3333)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AlwNI  (3026)
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  (2547)
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  (2481)
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.
BsrFI  (2462)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
NmeAIII  (2400)
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).
ScaI  (2066)
1 site
A G T A C T T C A T G A
XmnI  (1947)
1 site
G A A N N N N T T C C T T N N N N A A G
SmaI  (3798)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BspDI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoRV  (239)
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.
PsiI  (409)
1 site
T T A T A A A A T A T T
StyI  (568)
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  (606)
1 site
G C T A G C C G A T C G
BmtI  (610)
1 site
G C T A G C C G A T C G
SphI  (614)
1 site
G C A T G C C G T A C G
BsmI  (775)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BclI  (778)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BsiWI  (784)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
MluI  (872)
1 site
A C G C G T T G C G C A
BstBI  (1172)
1 site
T T C G A A A A G C T T
Acc65I  (1200)
1 site
G G T A C C C C A T G G
BsgI  (1202)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

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).
KpnI  (1204)
1 site
G G T A C C C C A T G G
BamHI  (1206)
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.
PshAI  (1216)
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.
SalI  (1219)
1 site
G T C G A C C A G C T G
AccI  (1220)
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.
HindIII  (1225)
1 site
A A G C T T T T C G A A
NotI  (1232)
1 site
G C G G C C G C C G C C G G C G
Eco53kI  (1243)
1 site
G A G C T C C T C G A G
BanII  (1245)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BseRI  (1245)
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.
SacI  (1245)
1 site
G A G C T C C T C G A G
DraIII  (1302)
1 site
C A C N N N G T G G T G N N N C