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pCMV-GLuc 2

Control vector with a strong constitutive CMV promoter for expressing secreted Gaussia luciferase in mammalian cells.

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pCMV-GLuc 2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  New England Biolabs
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SgrDI (5800) SspI (5683) ScaI (5359) PciI (3986) BstZ17I (3607) BsmI (3555) PfoI (3410) BstBI (3317) RsrII (3151) BssHII (3032) BglII (12) MfeI (161) Bpu10I (180) MluI (228) SpeI (249) NdeI (484) SnaBI (590) HindIII (889) Acc65I (895) KpnI (899) BamHI (907) BbsI (996) SacII (1051) NotI (1479) XbaI (1535) DraIII (1901) SexAI * (2192) BseRI (2421) StuI (2424) AvrII (2425) AvaI - BsoBI - TspMI - XmaI (2446) BmeT110I (2447) SmaI (2448) BclI * (2476) KasI (2634) NarI (2635) SfoI (2636) PluTI (2638) PstI (2688) PflFI - Tth111I (2753) pCMV-GLuc 2 5802 bp
SgrDI  (5800)
1 site
C G T C G A C G G C A G C T G C
SspI  (5683)
1 site
A A T A T T T T A T A A
ScaI  (5359)
1 site
A G T A C T T C A T G A
PciI  (3986)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3607)
1 site
G T A T A C C A T A T G
BsmI  (3555)
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.
PfoI  (3410)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstBI  (3317)
1 site
T T C G A A A A G C T T
RsrII  (3151)
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.
BssHII  (3032)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BglII  (12)
1 site
A G A T C T T C T A G A
MfeI  (161)
1 site
C A A T T G G T T A A C
Bpu10I  (180)
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.
MluI  (228)
1 site
A C G C G T T G C G C A
SpeI  (249)
1 site
A C T A G T T G A T C A
NdeI  (484)
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  (590)
1 site
T A C G T A A T G C A T
HindIII  (889)
1 site
A A G C T T T T C G A A
Acc65I  (895)
1 site
G G T A C C C C A T G G
KpnI  (899)
1 site
G G T A C C C C A T G G
BamHI  (907)
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.
BbsI  (996)
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.
SacII  (1051)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
NotI  (1479)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1535)
1 site
T C T A G A A G A T C T
DraIII  (1901)
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.
SexAI  (2192)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BseRI  (2421)
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.
StuI  (2424)
1 site
A G G C C T T C C G G A
AvrII  (2425)
1 site
C C T A G G G G A T C C
AvaI  (2446)
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  (2446)
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  (2446)
1 site
C C C G G G G G G C C C
XmaI  (2446)
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.
BmeT110I  (2447)
1 site
C Y C G R G G R G C Y C
SmaI  (2448)
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  (2476)
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.
KasI  (2634)
1 site
G G C G C C C C G C G G
NarI  (2635)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2636)
1 site
G G C G C C C C G C G G
PluTI  (2638)
1 site
G G C G C C C C G C G