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Plasmid Files

pIEx™-1

Vector for high-level expression in insect cells of proteins with a cleavable N-terminal 10xHis-S-Tag cassette plus a C-terminal HSV tag.

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pIEx-1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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BspQI - SapI (3648) PciI (3531) DrdI (3429) PspFI (3231) BseYI (3227) AhdI (2643) BsaI (2577) BpmI (2574) NmeAIII (2496) ScaI (2162) XmnI (2043) BspDI * - ClaI * (207) EcoRV (239) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BclI * (778) BsiWI (784) MluI (872) BtgI - NcoI (1082) XcmI (1120) PflMI (1164) PshAI (1238) BseRI (1242) BamHI (1260) MfeI (1266) Eco53kI (1275) BanII - SacI (1277) BsrGI (1280) AscI (1288) PstI - SbfI (1298) SalI (1300) Acc65I (1306) AgeI (1309) KpnI (1310) BstBI (1315) HindIII (1318) NotI (1325) BstZ17I (1340) PmlI (1345) BbsI (1379) PaeR7I - XhoI (1387) DraIII (1398) Bsu36I (1408) PacI (1437) PflFI - Tth111I (1661) BtgZI (1684) XbaI (1716) ZraI (1722) AatII (1724) pIEx™-1 3897 bp
BspQI  (3648)
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  (3648)
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  (3531)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (3429)
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  (3231)
1 site
C C C A G C G G G T C G
BseYI  (3227)
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.
AhdI  (2643)
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  (2577)
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  (2574)
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.
NmeAIII  (2496)
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  (2162)
1 site
A G T A C T T C A T G A
XmnI  (2043)
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.
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
BtgI  (1082)
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  (1082)
1 site
C C A T G G G G T A C C
XcmI  (1120)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PflMI  (1164)
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.
PshAI  (1238)
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.