pRL-TK

Mammalian co-reporter vector for the weak constitutive expression of wild-type Renilla luciferase.

Sequence Author: Promega

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BglII (1) AhdI (3146) BsrFI (3061) NmeAIII (2999) FspI (2923) PvuI (2777) SspI (2341) BamHI (2223) BsaBI * (2222) BspDI - ClaI (2216) MfeI (2130) HincII - HpaI (2121) MscI (128) BspQI - SapI (137) BssHII (241) TspMI - XmaI (282) SmaI (284) KasI (289) NarI (290) SfoI (291) PluTI (293) SacII (299) BanII (313) EcoO109I - PpuMI (319) AvrII (322) NcoI (330) AccI (342) BstZ17I (343) PvuII (531) EcoRI (649) MluI (713) HindIII (760) BfuAI - BspMI (816) NheI (1024) BmtI (1028) BstAPI (1166) PciI (1216) XcmI (1683) BsrGI (1732) XbaI (1971) EagI - NotI (1978) pRL-TK 4045 bp
BglII  (1)
1 site
A G A T C T T C T A G A
AhdI  (3146)
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  (3061)
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.
NmeAIII  (2999)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
FspI  (2923)
1 site
T G C G C A A C G C G T
PvuI  (2777)
1 site
C G A T C G G C T A G C
SspI  (2341)
1 site
A A T A T T T T A T A A
BamHI  (2223)
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.
BsaBI  (2222)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BspDI  (2216)
1 site
A T C G A T T A G C T A
ClaI  (2216)
1 site
A T C G A T T A G C T A
MfeI  (2130)
1 site
C A A T T G G T T A A C
HincII  (2121)
1 site
G T Y R A C C A R Y T G
HpaI  (2121)
1 site
G T T A A C C A A T T G
MscI  (128)
1 site
T G G C C A A C C G G T
BspQI  (137)
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  (137)
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.
BssHII  (241)
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.
TspMI  (282)
1 site
C C C G G G G G G C C C
XmaI  (282)
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  (284)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KasI  (289)
1 site
G G C G C C C C G C G G
NarI  (290)
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  (291)
1 site
G G C G C C C C G C G G
PluTI  (293)
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.
SacII  (299)
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.
BanII  (313)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
EcoO109I  (319)
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.
PpuMI  (319)
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.
AvrII  (322)
1 site
C C T A G G G G A T C C
NcoI  (330)
1 site
C C A T G G G G T A C C
AccI  (342)
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.
BstZ17I  (343)
1 site
G T A T A C C A T A T G
PvuII  (531)
1 site
C A G C T G G T C G A C
EcoRI  (649)
1 site
G A A T T C C T T A A G
MluI  (713)
1 site
A C G C G T T G C G C A
HindIII  (760)
1 site
A A G C T T T T C G A A
BfuAI  (816)
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  (816)
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.
NheI  (1024)
1 site
G C T A G C C G A T C G
BmtI  (1028)
1 site
G C T A G C C G A T C G
BstAPI  (1166)
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.
PciI  (1216)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
XcmI  (1683)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BsrGI  (1732)
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.
XbaI  (1971)
1 site
T C T A G A A G A T C T
EagI  (1978)
1 site
C G G C C G G C C G G C
NotI  (1978)
1 site
G C G G C C G C C G C C G G C G
Rluc
1034 .. 1969  =  936 bp
311 amino acids  =  36.0 kDa
Product: luciferase from the anthozoan coelenterate Renilla reniformis (sea pansy)
Rluc
1034 .. 1969  =  936 bp
311 amino acids  =  36.0 kDa
Product: luciferase from the anthozoan coelenterate Renilla reniformis (sea pansy)
AmpR
2359 .. 3219  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2359 .. 2427  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2359 .. 3219  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2428 .. 3219  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2359 .. 3219  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
HSV TK promoter
8 .. 759  =  752 bp
herpes simplex virus thymidine kinase promoter
HSV TK promoter
8 .. 759  =  752 bp
herpes simplex virus thymidine kinase promoter
ori
3390 .. 3978  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3390 .. 3978  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
chimeric intron
829 .. 961  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
chimeric intron
829 .. 961  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
SV40 poly(A) signal
2000 .. 2121  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2000 .. 2121  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2254 .. 2358  =  105 bp
AmpR promoter
2254 .. 2358  =  105 bp
T7 promoter
1006 .. 1024  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1006 .. 1024  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  2359 .. 3219  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  332 .. 796  =  465 bp
ORF:  154 amino acids  =  17.4 kDa
ORF:  1034 .. 1969  =  936 bp
ORF:  311 amino acids  =  36.0 kDa
ORF:  94 .. 540  =  447 bp
ORF:  148 amino acids  =  16.0 kDa
ORF:  2823 .. 3089  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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