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pGL4.13[luc2/SV40]

SV40 promoter-containing internal control vector for strong constitutive expression of luciferase.

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pGL4.13[luc2 SV40] Sequence and MappGL4.13[luc2 SV40].dna
Map and Sequence File   
Sequence Author:  Promega
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 AarI - BfuAI - BspMI (4611) RVprimer3 (4590 .. 4609) BsmBI (4438) SpeI (4425) BstZ17I (4106) SacII (3990) PvuI (3966) Bsu36I (3952) AhdI (3596) BstEII (3521) BstXI - PstI (3518) NotI (3494) AlwNI (3090) ApaLI (2988) BciVI (2877) AflIII - PciI (2674) RVprimer4 (2475 .. 2494) PshAI (2489) SalI (2424) BamHI (2418) MfeI (2325) BglI - SfiI (8) Eco53kI (23) SacI (25) NheI (27) BmtI (31) AbsI - PaeR7I - PspXI - XhoI (33) EcoRV (41) BglII (45) BglI - SfiI (401) StuI (447) AvrII (448) HindIII (464) PspOMI (525) ApaI (529) MreI - SgrAI (562) BsrGI (989) BbvCI (1210) KasI (1446) NarI (1447) SfoI (1448) PluTI (1450) BlpI (1453) AgeI (1630) DraIII (1645) BpmI (1879) XbaI (2156) FseI (2175) ApoI (2261) PsiI (2296) pGL4.13[luc2/SV40] 4641 bp
AarI  (4611)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BfuAI  (4611)
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  (4611)
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.
BsmBI  (4438)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
SpeI  (4425)
1 site
A C T A G T T G A T C A
BstZ17I  (4106)
1 site
G T A T A C C A T A T G
SacII  (3990)
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.
PvuI  (3966)
1 site
C G A T C G G C T A G C
Bsu36I  (3952)
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.
AhdI  (3596)
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.
BstEII  (3521)
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.
BstXI  (3518)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
PstI  (3518)
1 site
C T G C A G G A C G T C
NotI  (3494)
1 site
G C G G C C G C C G C C G G C G
AlwNI  (3090)
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.
ApaLI  (2988)
1 site
G T G C A C C A C G T G
BciVI  (2877)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
AflIII  (2674)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2674)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PshAI  (2489)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SalI  (2424)
1 site
G T C G A C C A G C T G
BamHI  (2418)
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.
MfeI  (2325)
1 site
C A A T T G G T T A A C
BglI  (8)
2 sites
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.
SfiI  (8)
2 sites
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
Eco53kI  (23)
1 site
G A G C T C C T C G A G
SacI  (25)
1 site
G A G C T C C T C G A G
NheI  (27)
1 site
G C T A G C C G A T C G
BmtI  (31)
1 site
G C T A G C C G A T C G
AbsI  (33)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (33)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (33)
1 site
V C T C G A G B B G A G C T C V
XhoI  (33)
1 site
C T C G A G G A G C T C
EcoRV  (41)
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.
BglII  (45)
1 site
A G A T C T T C T A G A
BglI  (401)
2 sites
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.
SfiI  (401)
2 sites
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
StuI  (447)
1 site
A G G C C T T C C G G A
AvrII  (448)
1 site
C C T A G G G G A T C C
HindIII  (464)
1 site
A A G C T T T T C G A A
PspOMI  (525)
1 site
G G G C C C C C C G G G
ApaI  (529)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
MreI  (562)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (562)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
BsrGI  (989)
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.
BbvCI  (1210)
1 site
C C T C A G C G G A G T C G
KasI  (1446)
1 site
G G C G C C C C G C G G
NarI  (1447)
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  (1448)
1 site
G G C G C C C C G C G G
PluTI  (1450)
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.
BlpI  (1453)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
AgeI  (1630)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
DraIII  (1645)
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.
BpmI  (1879)
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.
XbaI  (2156)
1 site
T C T A G A A G A T C T
FseI  (2175)
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.
ApoI  (2261)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
PsiI  (2296)
1 site
T T A T A A A A T A T T
RVprimer3
20-mer  /  50% GC
1 binding site
4590 .. 4609  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
2475 .. 2494  =  20 annealed bases
Tm  =  61°C
luciferase
499 .. 2151  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
499 .. 2151  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
AmpR
3523 .. 4383  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3523 .. 4314  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3523 .. 4383  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4315 .. 4383  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3523 .. 4383  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2735 .. 3323  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2735 .. 3323  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 promoter
106 .. 463  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
106 .. 463  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2195 .. 2316  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2195 .. 2316  =  122 bp
SV40 polyadenylation signal
pause site
4550 .. 4641  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
4550 .. 4641  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
poly(A) signal
4488 .. 4536  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4488 .. 4536  =  49 bp
synthetic polyadenylation signal
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