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pIEx™-5

Vector for high-level expression in insect cells of secreted proteins with a C-terminal S-Tag-8xHis cassette.

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pIEx-5.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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SmaI (3840) TspMI - XmaI (3838) BspQI - SapI (3594) PciI (3477) DrdI (3375) PspFI (3177) BseYI (3173) AlwNI (3068) AhdI (2589) BsaI (2523) BpmI (2520) ScaI (2108) XmnI (1989) BspDI * - ClaI * (207) EcoRV (239) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BclI * (778) BseRI (781) BsiWI (784) MluI (872) BsrGI (1085) BbsI (1092) BsaBI (1111) BbvCI - Bpu10I (1133) BtgI - NcoI (1145) HpaI (1157) BmgBI (1162) Eco53kI (1170) BanII - SacI (1172) BamHI (1176) MfeI (1182) BglII (1190) BfuAI - BspMI (1197) AscI (1198) PstI - SbfI (1208) SalI (1210) AccI (1211) Acc65I (1216) AgeI (1219) KpnI (1220) BstBI (1225) HindIII (1228) NotI (1235) PaeR7I - PspXI - XhoI (1312) DraIII (1344) Bsu36I (1354) PacI (1383) PflFI - Tth111I (1607) XbaI (1662) ZraI (1668) AatII (1670) pIEx™-5 3843 bp
SmaI  (3840)
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  (3838)
1 site
C C C G G G G G G C C C
XmaI  (3838)
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  (3594)
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  (3594)
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  (3477)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (3375)
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.
PspFI  (3177)
1 site
C C C A G C G G G T C G
BseYI  (3173)
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  (3068)
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  (2589)
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  (2523)
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  (2520)
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.
ScaI  (2108)
1 site
A G T A C T T C A T G A
XmnI  (1989)
1 site
G A A N N N N T T C C T T N N N N A A G
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
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.
BseRI  (781)
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.
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
BsrGI  (1085)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BbsI  (1092)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsaBI  (1111)
1 site
G A T N N N N A T C C T A N N N N T A G
BbvCI  (1133)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1133)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
BtgI  (1145)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1145)
1 site
C C A T G G G G T A C C
HpaI  (1157)
1 site
G T T A A C C A A T T G
BmgBI  (1162)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
Eco53kI  (1170)
1 site
G A G C T C C T C G A G
BanII  (1172)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (1172)
1 site
G A G C T C C T C G A G
BamHI  (1176)
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.
MfeI  (1182)
1 site
C A A T T G G T T A A C
BglII  (1190)
1 site
A G A T C T T C T A G A
BfuAI  (1197)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1197)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
AscI  (1198)
1 site
G G C G C G C C C C G C G C G G
PstI  (1208)
1 site
C T G C A G G A C G T C
SbfI  (1208)
1 site
C C T G C A G G G G A C G T C C
SalI  (1210)
1 site
G T C G A C C A G C T G
AccI  (1211)
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.
Acc65I  (1216)
1 site
G G T A C C C C A T G G
AgeI  (1219)
1 site
A C C G G T T G G C C A
KpnI  (1220)
1 site
G G T A C C C C A T G G
BstBI  (1225)
1 site
T T C G A A A A G C T T
HindIII  (1228)
1 site
A A G C T T T T C G A A
NotI  (1235)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1312)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1312)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1312)
1 site
C T C G A G G A G C T