pSF-PromMCS-RLuc

Vector for measuring promoter activity using Renilla luciferase.

Sequence Author: Oxford Genetics

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HomePlasmidsLuciferase VectorspSF-PromMCS-RLuc
AsiSI - PvuI (5) PmeI (4591) RsrII (4284) BsrDI (4001) PflFI - Tth111I (3886) RBS PmeI (3543) PpuMI - SanDI (3477) AscI (3386) FseI (3240) NaeI (3238) NgoMIV (3236) SwaI (3134) BspHI (2965) AlwNI (2661) BssSI (2418) SwaI (2241) 5' β-globin insulator BglII (232) ScaI (241) pause site AarI (306) SalI (337) AccI (338) SpeI (345) TspMI - XmaI (353) SmaI (355) PspOMI (361) ApaI (365) BstBI (370) BglII (377) EagI - NotI (419) HindIII (430) Eco53kI (440) SacI (442) EcoRI (446) Acc65I (458) KpnI (462) BstAPI (656) Bpu10I (1116) BseRI - BsgI (1405) stop codons BspDI - ClaI (1442) BamHI (1451) StuI (1461) NheI (1467) BmtI (1471) stop codons HpaI (1637) MfeI (1646) BglI (1806) PstI - SbfI (1983) 3' β-globin insulator BstEII (2089) PacI (2115) pSF-PromMCS- RLuc 4710 bp
AsiSI  (5)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (5)
1 site		 C G A T C G G C T A G C
PmeI  (4591)
2 sites		 G T T T A A A C C A A A T T T G
RsrII  (4284)
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.
BsrDI  (4001)
1 site		 G C A A T G N N C G T T A C
Sticky ends from different BsrDI sites may not be compatible.
PflFI  (3886)
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  (3886)
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.
PmeI  (3543)
2 sites		 G T T T A A A C C A A A T T T G
PpuMI  (3477)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
SanDI  (3477)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different SanDI sites may not be compatible.
AscI  (3386)
1 site		 G G C G C G C C C C G C G C G G
FseI  (3240)
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.
NaeI  (3238)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (3236)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
SwaI  (3134)
2 sites		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
BspHI  (2965)
1 site		 T C A T G A A G T A C T
AlwNI  (2661)
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.
BssSI  (2418)
1 site		 C A C G A G G T G C T C
SwaI  (2241)
2 sites		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
BglII  (232)
2 sites		 A G A T C T T C T A G A
ScaI  (241)
1 site		 A G T A C T T C A T G A
AarI  (306)
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.
SalI  (337)
1 site		 G T C G A C C A G C T G
AccI  (338)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SpeI  (345)
1 site		 A C T A G T T G A T C A
TspMI  (353)
1 site		 C C C G G G G G G C C C
XmaI  (353)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (355)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
PspOMI  (361)
1 site		 G G G C C C C C C G G G
ApaI  (365)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BstBI  (370)
1 site		 T T C G A A A A G C T T
BglII  (377)
2 sites		 A G A T C T T C T A G A
EagI  (419)
1 site		 C G G C C G G C C G G C
NotI  (419)
1 site		 G C G G C C G C C G C C G G C G
HindIII  (430)
1 site		 A A G C T T T T C G A A
Eco53kI  (440)
1 site		 G A G C T C C T C G A G
SacI  (442)
1 site		 G A G C T C C T C G A G
EcoRI  (446)
1 site		 G A A T T C C T T A A G
Acc65I  (458)
1 site		 G G T A C C C C A T G G
KpnI  (462)
1 site		 G G T A C C C C A T G G
BstAPI  (656)
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.
Bpu10I  (1116)
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.
BseRI  (1405)
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.
BsgI  (1405)
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).
BspDI  (1442)
1 site		 A T C G A T T A G C T A
ClaI  (1442)
1 site		 A T C G A T T A G C T A
BamHI  (1451)
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.
StuI  (1461)
1 site		 A G G C C T T C C G G A
NheI  (1467)
1 site		 G C T A G C C G A T C G
BmtI  (1471)
1 site		 G C T A G C C G A T C G
HpaI  (1637)
1 site		 G T T A A C C A A T T G
MfeI  (1646)
1 site		 C A A T T G G T T A A C
BglI  (1806)
1 site		 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.
PstI  (1983)
1 site		 C T G C A G G A C G T C
SbfI  (1983)
1 site		 C C T G C A G G G G A C G T C C
BstEII  (2089)
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.
PacI  (2115)
1 site		 T T A A T T A A A A T T A A T T
RLuc
468 .. 1406  =  939 bp
312 amino acids  =  36.1 kDa
Product: Renilla luciferase
codon-optimized synthetic gene
RLuc
468 .. 1406  =  939 bp
312 amino acids  =  36.1 kDa
Product: Renilla luciferase
codon-optimized synthetic gene
NeoR/KanR
3640 .. 4434  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3640 .. 4434  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2306 .. 2894  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2306 .. 2894  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rrnG terminator
1828 .. 1964  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
1828 .. 1964  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
4447 .. 4583  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
4447 .. 4583  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
1516 .. 1637  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1516 .. 1637  =  122 bp
SV40 polyadenylation signal
pause site
245 .. 336  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
245 .. 336  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
5' β-globin insulator
18 .. 89  =  72 bp
insulator upstream of the human β-globin locus (Farrell et al., 2002)
5' β-globin insulator
18 .. 89  =  72 bp
insulator upstream of the human β-globin locus (Farrell et al., 2002)
3' β-globin insulator
2009 .. 2080  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
3' β-globin insulator
2009 .. 2080  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
T7 terminator
1752 .. 1798  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1752 .. 1798  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
MCS
337 .. 374  =  38 bp
multiple cloning site
MCS
337 .. 374  =  38 bp
multiple cloning site
stop codons
1426 .. 1436  =  11 bp
stop codons in all three reading frames
stop codons
1426 .. 1436  =  11 bp
stop codons in all three reading frames
stop codons
1474 .. 1484  =  11 bp
stop codons in all three reading frames
stop codons
1474 .. 1484  =  11 bp
stop codons in all three reading frames
RBS
454 .. 459  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
454 .. 459  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
3627 .. 3632  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
3627 .. 3632  =  6 bp
Shine-Dalgarno ribosome binding site
ORF:  3640 .. 4434  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1853 .. 2284  =  432 bp
ORF:  143 amino acids  =  16.3 kDa
ORF:  3812 .. 4198  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  4472 .. 196  =  435 bp
ORF:  144 amino acids  =  16.1 kDa
ORF:  468 .. 1406  =  939 bp
ORF:  312 amino acids  =  36.1 kDa
ORF:  450 .. 1322  =  873 bp
ORF:  290 amino acids  =  34.7 kDa
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pGL4.44[luc2P AP1 RE Hygro]
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pGL2-Enhancer
pGL4.45[luc2P ISRE Hygro]
pMetLuc2-Control
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pGL2-Promoter
pGL4.47[luc2P SIE Hygro]
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pGL4.73[hRluc SV40]
pNL2.1[Nluc Hygro]
pTK-Cypridina Luc
pGL4.15[luc2P Hygro]
pGL4.74[hRluc TK]
pNL2.2[NlucP Hygro]
pTK-Gaussia Luc
pGL4.16[luc2CP Hygro]
pGL4.75[hRluc CMV]
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pTK-Gaussia-Dura Luc
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