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Plasmid Files

pRL-null

Promoterless mammalian vector encoding wild-type Renilla luciferase.

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pRL-null Sequence and MappRL-null.dna
Map and Sequence File   
Sequence Author:  Promega
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 Eco53kI (10) PaeR7I - XhoI (5) BglII (1) DrdI (3207) PspFI (3009) BseYI (3005) AlwNI (2900) AhdI (2421) BpmI (2352) BsrFI (2336) BglI (2303) NmeAIII (2274) FspI (2198) PvuI (2052) BanII - SacI (12) HindIII (15) NdeI (24) NsiI (30) SphI (32) SpeI (34) KasI (40) NarI (41) SfoI (42) PluTI (44) SalI (47) AccI (48) MluI (51) EcoRI (58) TspMI - XmaI (65) SmaI (67) PstI (77) BfuAI - BspMI (91) AflII (264) T7 promoter NheI (299) BmtI (303) BstBI (315) BstAPI (441) PciI (491) XcmI (958) BsrGI (1007) BsaAI (1041) XbaI (1246) EagI - NotI (1253) HpaI (1396) MfeI (1405) BspDI - ClaI (1491) BsaBI * (1497) BamHI (1498) SspI (1616) pRL-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.
BpmI  (2352)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BsrFI  (2336)
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.
BglI  (2303)
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.
NmeAIII  (2274)
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  (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
BanII  (12)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
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.
NsiI  (30)
1 site
A T G C A T T A C G T A
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.
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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.
AflII  (264)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
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
BstBI  (315)
1 site
T T C G A A A A G C T T
BstAPI  (441)
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  (491)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
XcmI  (958)
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  (1007)
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.
BsaAI  (1041)
1 site
Y A C G T R R T G C A Y
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
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
BsaBI  (1497)
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.
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.
SspI  (1616)
1 site
A A T A T T T T A T A A
Rluc
309 .. 1244  =  936 bp
311 amino acids  =  36.0 kDa
Product: luciferase from the anthozoan coelenterate
Renilla reniformis (sea pansy)
Rluc
309 .. 1244  =  936 bp
311 amino acids  =  36.0 kDa
Product: luciferase from the anthozoan coelenterate
Renilla reniformis (sea pansy)
AmpR
1634 .. 2494  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
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