phRL-null

Promoterless mammalian vector encoding humanized Renilla luciferase.

Sequence Author: Promega

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Eco53kI (10) PaeR7I - XhoI (5) BglII (1) DrdI (3207) PspFI (3009) BseYI (3005) AlwNI (2900) AhdI (2421) BsrFI (2336) AseI (2246) FspI (2198) PvuI (2052) SspI (1616) SacI (12) HindIII (15) NdeI (24) NspI - SphI (32) SpeI (34) KasI (40) NarI (41) SfoI (42) PluTI (44) SalI (47) AccI (48) AflIII - MluI (51) EcoRI (58) TspMI - XmaI (65) SmaI (67) PstI (77) BfuAI - BspMI (91) BbsI (175) AflII (264) T7 promoter NheI (299) BmtI (303) NcoI (307) BstAPI - DraIII (497) PfoI * (589) EcoRV (794) Bsu36I (895) NruI (970) ZraI (1004) PflFI - Tth111I (1005) AatII (1006) PasI (1083) EcoO109I (1154) XbaI (1246) EagI - NotI (1253) PsiI (1376) HpaI (1396) MfeI (1405) BspDI - ClaI (1491) BamHI (1498) phRL-null 3320 bp
Eco53kI  (10)
1 site
G A G C T C C T C G A G
PaeR7I  (5)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (5)
1 site
C T C G A G G A G C T C
BglII  (1)
1 site
A G A T C T T C T A G A
DrdI  (3207)
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.
PspFI  (3009)
1 site
C C C A G C G G G T C G
BseYI  (3005)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2900)
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.
AhdI  (2421)
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.
BsrFI  (2336)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
AseI  (2246)
1 site
A T T A A T T A A T T A
FspI  (2198)
1 site
T G C G C A A C G C G T
PvuI  (2052)
1 site
C G A T C G G C T A G C
SspI  (1616)
1 site
A A T A T T T T A T A A
SacI  (12)
1 site
G A G C T C C T C G A G
HindIII  (15)
1 site
A A G C T T T T C G A A
NdeI  (24)
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.
NspI  (32)
1 site
R C A T G Y Y G T A C R
SphI  (32)
1 site
G C A T G C C G T A C G
SpeI  (34)
1 site
A C T A G T T G A T C A
KasI  (40)
1 site
G G C G C C C C G C G G
NarI  (41)
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  (42)
1 site
G G C G C C C C G C G G
PluTI  (44)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SalI  (47)
1 site
G T C G A C C A G C T G
AccI  (48)
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.
AflIII  (51)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (51)
1 site
A C G C G T T G C G C A
EcoRI  (58)
1 site
G A A T T C C T T A A G
TspMI  (65)
1 site
C C C G G G G G G C C C
XmaI  (65)
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  (67)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PstI  (77)
1 site
C T G C A G G A C G T C
BfuAI  (91)
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  (91)
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.
BbsI  (175)
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.
AflII  (264)
1 site
C T T A A G G A A T T C
NheI  (299)
1 site
G C T A G C C G A T C G
BmtI  (303)
1 site
G C T A G C C G A T C G
NcoI  (307)
1 site
C C A T G G G G T A C C
BstAPI  (497)
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  (497)
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.
PfoI  (589)
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  (794)
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.
Bsu36I  (895)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
NruI  (970)
1 site
T C G C G A A G C G C T
ZraI  (1004)
1 site
G A C G T C C T G C A G
PflFI  (1005)
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  (1005)
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  (1006)
1 site
G A C G T C C T G C A G
PasI  (1083)
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.
EcoO109I  (1154)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
XbaI  (1246)
1 site
T C T A G A A G A T C T
EagI  (1253)
1 site
C G G C C G G C C G G C
NotI  (1253)
1 site
G C G G C C G C C G C C G G C G
PsiI  (1376)
1 site
T T A T A A A A T A T T
HpaI  (1396)
1 site
G T T A A C C A A T T G
MfeI  (1405)
1 site
C A A T T G G T T A A C
BspDI  (1491)
1 site
A T C G A T T A G C T A
ClaI  (1491)
1 site
A T C G A T T A G C T A
BamHI  (1498)
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.
hRluc
309 .. 1244  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
309 .. 1244  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
AmpR
1634 .. 2494  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1634 .. 1702  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1634 .. 2494  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1703 .. 2494  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1634 .. 2494  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2665 .. 3253  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2665 .. 3253  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
chimeric intron
104 .. 236  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
chimeric intron
104 .. 236  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
SV40 poly(A) signal
1275 .. 1396  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1275 .. 1396  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1529 .. 1633  =  105 bp
AmpR promoter
1529 .. 1633  =  105 bp
MCS
1 .. 78  =  78 bp
multiple cloning site
MCS
1 .. 78  =  78 bp
multiple cloning site
T7 promoter
281 .. 299  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
281 .. 299  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1634 .. 2494  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  309 .. 1244  =  936 bp
ORF:  311 amino acids  =  36.0 kDa
ORF:  2098 .. 2364  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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