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pGL4.75[hRluc/CMV]

Co-transfection vector for expressing Renilla luciferase under control of the strong CMV promoter.

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pGL4.75[hRluc CMV].dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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BsgI (4279) AarI - BfuAI - BspMI (4251) RVprimer3 (4230 .. 4249) BsmBI (4078) BstZ17I (3746) SacII (3630) PvuI (3606) AhdI (3236) BstEII (3161) BstXI - PstI (3158) AleI (3156) NotI (3134) BspHI (3034) AlwNI (2730) BaeGI - Bme1580I (2632) ApaLI (2628) BciVI (2517) DrdI (2422) NspI (2318) AflIII - PciI (2314) BspQI - SapI (2198) AfeI (2190) BbsI (2143) SfiI (8) BsrGI (109) AseI (173) NdeI (400) BtgZI (500) BsaAI - SnaBI (506) Eco53kI (732) SacI (734) SfiI (818) AvaI - BsoBI (880) BmeT110I (881) BstAPI - DraIII (1047) NsiI (1078) PfoI * (1139) EcoRV (1344) XmnI (1393) BsaI (1399) NruI (1520) PflFI - Tth111I (1555) PasI (1633) XbaI (1796) FseI (1815) ApoI (1901) AanI - PsiI (1936) HpaI (1956) MfeI (1965) BamHI (2058) SalI (2064) PshAI (2129) RVprimer4 (2115 .. 2134) pGL4.75[hRluc/CMV] 4281 bp
BsgI  (4279)
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).
AarI  (4251)
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.
BfuAI  (4251)
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  (4251)
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.
BsmBI  (4078)
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.
BstZ17I  (3746)
1 site
G T A T A C C A T A T G
SacII  (3630)
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.
PvuI  (3606)
1 site
C G A T C G G C T A G C
AhdI  (3236)
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.
BstEII  (3161)
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.
BstXI  (3158)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
PstI  (3158)
1 site
C T G C A G G A C G T C
AleI  (3156)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (3134)
1 site
G C G G C C G C C G C C G G C G
BspHI  (3034)
1 site
T C A T G A A G T A C T
AlwNI  (2730)
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.
BaeGI  (2632)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (2632)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
ApaLI  (2628)
1 site
G T G C A C C A C G T G
BciVI  (2517)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
DrdI  (2422)
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.
NspI  (2318)
1 site
R C A T G Y Y G T A C R
AflIII  (2314)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2314)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2198)
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  (2198)
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.
AfeI  (2190)
1 site
A G C G C T T C G C G A
BbsI  (2143)
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.
SfiI  (8)
2 sites
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BsrGI  (109)
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.
AseI  (173)
1 site
A T T A A T T A A T T A
NdeI  (400)
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.
BtgZI  (500)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (506)
1 site
Y A C G T R R T G C A Y
SnaBI  (506)
1 site
T A C G T A A T G C A T
Eco53kI  (732)
1 site
G A G C T C C T C G A G
SacI  (734)
1 site
G A G C T C C T C G A G
SfiI  (818)
2 sites
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvaI  (880)
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  (880)
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.
BmeT110I  (881)
1 site
C Y C G R G G R G C Y C
BstAPI  (1047)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
DraIII  (1047)
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.
NsiI  (1078)
1 site
A T G C A T T A C G T A
PfoI  (1139)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
EcoRV  (1344)
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.
XmnI  (1393)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaI  (1399)
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.
NruI  (1520)
1 site
T C G C G A A G C G C T
PflFI  (1555)
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  (1555)
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.
PasI  (1633)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
XbaI  (1796)
1 site
T C T A G A A G A T C T