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Plasmid Files

pRL-TK

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

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pRL-TK Sequence and MappRL-TK.dna
Map and Sequence File   
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

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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
   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
   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
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