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pCMV-Gaussia-Dura Luc

Control vector for constitutive high-level expression of secreted Gaussia-Dura luciferase under control of the CMV promoter.

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pCMV-Gaussia-Dura Luc.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Pierce)
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AarI (5452) EcoRV (5158) ApaI (4818) PspOMI (4814) AflII (4805) DrdI (4472) MauBI (3876) ScaI (3359) FspI (3101) AfeI (2929) BstZ17I (2764) SpeI (1) NdeI (236) SnaBI (342) Eco53kI (568) SacI (570) BamHI (634) BsaBI * (676) BbsI (722) NotI (1205) BseRI (1770) AvrII (1774) TspMI - XmaI (1795) SmaI (1797) BclI * (1825) PflFI - Tth111I (1945) BsiWI (1959) RsrII (2019) BstEII (2037) EarI (2062) BsmBI (2295) DraIII (2474) NgoMIV (2575) NaeI (2577) BsmI (2712) pCMV-Gaussia-Dura Luc 5482 bp
AarI  (5452)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
EcoRV  (5158)
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.
ApaI  (4818)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4814)
1 site
G G G C C C C C C G G G
AflII  (4805)
1 site
C T T A A G G A A T T C
DrdI  (4472)
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.
MauBI  (3876)
1 site
C G C G C G C G G C G C G C G C
ScaI  (3359)
1 site
A G T A C T T C A T G A
FspI  (3101)
1 site
T G C G C A A C G C G T
AfeI  (2929)
1 site
A G C G C T T C G C G A
BstZ17I  (2764)
1 site
G T A T A C C A T A T G
SpeI  (1)
1 site
A C T A G T T G A T C A
NdeI  (236)
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.
SnaBI  (342)
1 site
T A C G T A A T G C A T
Eco53kI  (568)
1 site
G A G C T C C T C G A G
SacI  (570)
1 site
G A G C T C C T C G A G
BamHI  (634)
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.
BsaBI  (676)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BbsI  (722)
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.
NotI  (1205)
1 site
G C G G C C G C C G C C G G C G
BseRI  (1770)
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.
AvrII  (1774)
1 site
C C T A G G G G A T C C
TspMI  (1795)
1 site
C C C G G G G G G C C C
XmaI  (1795)
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.
SmaI  (1797)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BclI  (1825)
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.
PflFI  (1945)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1945)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsiWI  (1959)
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.
RsrII  (2019)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstEII  (2037)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
EarI  (2062)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI recognition sequence.
Sticky ends from different EarI sites may not be compatible.
BsmBI  (2295)
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.
DraIII  (2474)
1 site
C A C N N N G T G G