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Plasmid Files

pGLuc-Basic 2

Mammalian promoter reporter vector encoding secreted Gaussia luciferase.

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pGLuc-Basic 2 Sequence and MappGLuc-Basic 2.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 AatII (4957) SgrDI (4956) ZraI (4955) SspI (4839) ScaI (4515) TatI (4513) BglI (4155) BsaI (4096) AflIII - PciI (3142) BstZ17I (2763) BsmI (2711) PfoI (2566) BglII (12) EcoRI (20) EcoRV (32) PaeR7I - PspXI - XhoI (36) HindIII (45) Acc65I (51) KpnI (55) Eco53kI (59) SacI (61) BamHI (63) BbsI (152) NruI (181) SacII (207) NotI (635) XbaI (691) DraIII (1057) SexAI * (1348) BseRI (1577) StuI (1580) AvrII (1581) TspMI - XmaI (1602) SmaI (1604) BclI * (1632) KasI (1790) NarI (1791) SfoI (1792) PluTI (1794) PflFI - Tth111I (1909) BssHII (2188) RsrII (2307) BstBI (2473) pGLuc-Basic 2 4958 bp
AatII  (4957)
1 site
G A C G T C C T G C A G
SgrDI  (4956)
1 site
C G T C G A C G G C A G C T G C
ZraI  (4955)
1 site
G A C G T C C T G C A G
SspI  (4839)
1 site
A A T A T T T T A T A A
ScaI  (4515)
1 site
A G T A C T T C A T G A
TatI  (4513)
1 site
W G T A C W W C A T G W
BglI  (4155)
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.
BsaI  (4096)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AflIII  (3142)
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  (3142)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (2763)
1 site
G T A T A C C A T A T G
BsmI  (2711)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
PfoI  (2566)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BglII  (12)
1 site
A G A T C T T C T A G A
EcoRI  (20)
1 site
G A A T T C C T T A A G
EcoRV  (32)
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.
PaeR7I  (36)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (36)
1 site
V C T C G A G B B G A G C T C V
XhoI  (36)
1 site
C T C G A G G A G C T C
HindIII  (45)
1 site
A A G C T T T T C G A A
Acc65I  (51)
1 site
G G T A C C C C A T G G
KpnI  (55)
1 site
G G T A C C C C A T G G
Eco53kI  (59)
1 site
G A G C T C C T C G A G
SacI  (61)
1 site
G A G C T C C T C G A G
BamHI  (63)
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.
BbsI  (152)
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.
NruI  (181)
1 site
T C G C G A A G C G C T
SacII  (207)
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.
NotI  (635)
1 site
G C G G C C G C C G C C G G C G
XbaI  (691)
1 site
T C T A G A A G A T C T
DraIII  (1057)
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.
SexAI  (1348)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BseRI  (1577)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
StuI  (1580)
1 site
A G G C C T T C C G G A
AvrII  (1581)
1 site
C C T A G G G G A T C C
TspMI  (1602)
1 site
C C C G G G G G G C C C
XmaI  (1602)
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  (1604)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BclI  (1632)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
KasI  (1790)
1 site
G G C G C C C C G C G G
NarI  (1791)
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  (1792)
1 site
G G C G C C C C G C G G
PluTI  (1794)
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.
PflFI  (1909)
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  (1909)
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.
BssHII  (2188)
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.
RsrII  (2307)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstBI  (2473)
1 site
T T C G A A A A G C T T
AmpR
3962 .. 4822  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3962 .. 4753  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3962 .. 4822  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4754 .. 4822  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3962 .. 4822  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
1663 .. 2457  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1663 .. 2457  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
ori
3203 .. 3791  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3203 .. 3791  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hGLuc
76 .. 633  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
hGLuc
76 .. 633  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
f1 ori
824 .. 1252  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
824 .. 1252  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
1266 .. 1596  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
1266 .. 1596  =  331 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2631 .. 2752  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2631 .. 2752  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4823 .. 4927  =  105 bp
AmpR promoter
4823 .. 4927  =  105 bp
MCS
12 .. 68  =  57 bp
multiple cloning site
MCS
12 .. 68  =  57 bp
multiple cloning site
poly(A) signal
642 .. 690  =  49 bp
synthetic polyadenylation signal
poly(A) signal
642 .. 690  =  49 bp
synthetic polyadenylation signal
SV40 ori
1447 .. 1582  =  136 bp
SV40 origin of replication
SV40 ori
1447 .. 1582  =  136 bp
SV40 origin of replication
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