pGL4.82[hRluc Puro]

Promoterless vector for measuring the activity of promoter and enhancer sequences with a Renilla luciferase assay.

Sequence Author: Promega

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HomePlasmidsLuciferase VectorspGL4.82[hRluc Puro]
BsgI (4682) AarI - BfuAI - BspMI (4654) RVprimer3 (4633 .. 4652) poly(A) signal BsmBI (4481) SpeI (4468) BstZ17I (4149) SacII (4033) PvuI (4009) AhdI (3639) BstEII (3564) BstXI - PstI (3561) AleI (3559) NotI (3537) BspHI (3437) ApaLI (3031) BciVI (2920) BssSI (2890) DrdI (2825) AflIII - PciI (2717) AfeI (2593) BbsI (2546) RVprimer4 (2518 .. 2537) SalI (2467) BstBI (2453) AgeI (2388) BglI - SfiI (8) Acc65I (14) KpnI (18) Bpu10I (19) Eco53kI (23) SacI (25) NheI (27) BmtI (31) AbsI - PaeR7I - PspXI - XhoI (33) BglII (46) BglI - SfiI (59) HindIII (65) BglI (100) PfoI * (380) XmnI (634) BsaI (640) ZraI (795) PflFI - Tth111I (796) AatII (797) PasI (874) FseI (1056) ApoI (1142) AanI - PsiI (1177) HpaI (1197) MfeI (1206) BamHI (1299) FspI (1337) PvuII (1409) StuI (1732) BtgZI (1831) BlpI (2070) SgrAI (2238) pGL4.82[hRluc/Puro] 4684 bp
BsgI  (4682)
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  (4654)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI  (4654)
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  (4654)
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  (4481)
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.
BsmBI-v2 is an improved version of BsmBI.
SpeI  (4468)
1 site		 A C T A G T T G A T C A
BstZ17I  (4149)
1 site		 G T A T A C C A T A T G
SacII  (4033)
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  (4009)
1 site		 C G A T C G G C T A G C
AhdI  (3639)
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  (3564)
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  (3561)
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  (3561)
1 site		 C T G C A G G A C G T C
AleI  (3559)
1 site		 C A C N N N N G T G G T G N N N N C A C
NotI  (3537)
1 site		 G C G G C C G C C G C C G G C G
BspHI  (3437)
1 site		 T C A T G A A G T A C T
ApaLI  (3031)
1 site		 G T G C A C C A C G T G
BciVI  (2920)
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.
BssSI  (2890)
1 site		 C A C G A G G T G C T C
DrdI  (2825)
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.
AflIII  (2717)
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  (2717)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AfeI  (2593)
1 site		 A G C G C T T C G C G A
BbsI  (2546)
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.
SalI  (2467)
1 site		 G T C G A C C A G C T G
BstBI  (2453)
1 site		 T T C G A A A A G C T T
AgeI  (2388)
1 site		 A C C G G T T G G C C A
BglI  (8)
3 sites		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
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.
Acc65I  (14)
1 site		 G G T A C C C C A T G G
KpnI  (18)
1 site		 G G T A C C C C A T G G
Bpu10I  (19)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
Eco53kI  (23)
1 site		 G A G C T C C T C G A G
SacI  (25)
1 site		 G A G C T C C T C G A G
NheI  (27)
1 site		 G C T A G C C G A T C G
BmtI  (31)
1 site		 G C T A G C C G A T C G
AbsI  (33)
1 site		 C C T C G A G G G G A G C T C C
PaeR7I  (33)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (33)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (33)
1 site		 C T C G A G G A G C T C
BglII  (46)
1 site		 A G A T C T T C T A G A
BglI  (59)
3 sites		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
SfiI  (59)
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.
HindIII  (65)
1 site		 A A G C T T T T C G A A
BglI  (100)
3 sites		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
PfoI  (380)
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.
XmnI  (634)
1 site		 G A A N N N N T T C C T T N N N N A A G
BsaI  (640)
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.
ZraI  (795)
1 site		 G A C G T C C T G C A G
PflFI  (796)
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  (796)
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.
AatII  (797)
1 site		 G A C G T C C T G C A G
PasI  (874)
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.
FseI  (1056)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
ApoI  (1142)
1 site		 R A A T T Y Y T T A A R
ApoI is typically used at 50°C, but is 50% active at 37°C.
AanI  (1177)
1 site		 T T A T A A A A T A T T
PsiI  (1177)
1 site		 T T A T A A A A T A T T
HpaI  (1197)
1 site		 G T T A A C C A A T T G
MfeI  (1206)
1 site		 C A A T T G G T T A A C
BamHI  (1299)
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.
FspI  (1337)
1 site		 T G C G C A A C G C G T
PvuII  (1409)
1 site		 C A G C T G G T C G A C
StuI  (1732)
1 site		 A G G C C T T C C G G A
BtgZI  (1831)
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.
BlpI  (2070)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
SgrAI  (2238)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
RVprimer3
20-mer  /  50% GC
1 binding site
4633 .. 4652  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
2518 .. 2537  =  20 annealed bases
Tm  =  62°C
hRluc
100 .. 1035  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
100 .. 1035  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
AmpR
3566 .. 4426  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3566 .. 4357  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3566 .. 4426  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4358 .. 4426  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3566 .. 4426  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
1779 .. 2378  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1779 .. 2378