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Plasmid Files

pGL2-Basic

Promoterless vector for measuring the activity of promoter and enhancer sequences with a luciferase assay.

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pGL2-Basic Sequence and MappGL2-Basic.dna
Map and Sequence File   
Sequence Author:  Promega
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 KpnI (12) Acc65I (8) SmaI (3) TspMI - XmaI (1) GLprimer1 (5565 .. 5587) DraIII (5040) XmnI (4487) ScaI (4368) EaeI (4276) NmeAIII (4036) BsaI (3949) AhdI (3888) AlwNI (3411) PciI (2995) AfeI (2871) RVprimer4 (2796 .. 2815) Eco53kI (16) SacI (18) MluI (22) MCS NheI (28) BmtI (32) PaeR7I - XhoI (33) BglII (37) HindIII (47) GLprimer2 (77 .. 99) KasI (108) NarI (109) SfoI (110) PluTI (112) XbaI (123) PfoI * (183) BsiWI (230) BsrGI (566) EcoRI (663) BstEII (683) Bsu36I (689) SphI (739) XcmI (811) EcoO109I - PpuMI (1255) PacI (1400) EcoRV (1414) BspDI - ClaI (1441) BfuAI - BspMI (1474) SgrAI (1504) EcoNI (1693) PflMI (2044) StyI (2192) BsgI (2276) BsaBI * (2507) BamHI (2739) SalI (2745) AccI (2746) PshAI (2810) pGL2-Basic 5598 bp
KpnI  (12)
1 site
G G T A C C C C A T G G
Acc65I  (8)
1 site
G G T A C C C C A T G G
SmaI  (3)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (1)
1 site
C C C G G G G G G C C C
XmaI  (1)
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.
DraIII  (5040)
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.
XmnI  (4487)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (4368)
1 site
A G T A C T T C A T G A
EaeI  (4276)
1 site
Y G G C C R R C C G G Y
NmeAIII  (4036)
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).
BsaI  (3949)
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.
AhdI  (3888)
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.
AlwNI  (3411)
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.
PciI  (2995)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AfeI  (2871)
1 site
A G C G C T T C G C G A
Eco53kI  (16)
1 site
G A G C T C C T C G A G
SacI  (18)
1 site
G A G C T C C T C G A G
MluI  (22)
1 site
A C G C G T T G C G C A
NheI  (28)
1 site
G C T A G C C G A T C G
BmtI  (32)
1 site
G C T A G C C G A T C G
PaeR7I  (33)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (33)
1 site
C T C G A G G A G C T C
BglII  (37)
1 site
A G A T C T T C T A G A
HindIII  (47)
1 site
A A G C T T T T C G A A
KasI  (108)
1 site
G G C G C C C C G C G G
NarI  (109)
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  (110)
1 site
G G C G C C C C G C G G
PluTI  (112)
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.
XbaI  (123)
1 site
T C T A G A A G A T C T
PfoI  (183)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BsiWI  (230)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BsrGI  (566)
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.
EcoRI  (663)
1 site
G A A T T C C T T A A G
BstEII  (683)
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.
Bsu36I  (689)
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.
SphI  (739)
1 site
G C A T G C C G T A C G
XcmI  (811)
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.
EcoO109I  (1255)
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  (1255)
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.
PacI  (1400)
1 site
T T A A T T A A A A T T A A T T
EcoRV  (1414)
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.
BspDI  (1441)
1 site
A T C G A T T A G C T A
ClaI  (1441)
1 site
A T C G A T T A G C T A
BfuAI  (1474)
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  (1474)
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.
SgrAI  (1504)
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.
EcoNI  (1693)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PflMI  (2044)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
StyI  (2192)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BsgI  (2276)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaBI  (2507)
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  (2739)
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.
SalI  (2745)
1 site
G T C G A C C A G C T G
AccI  (2746)
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.
PshAI  (2810)
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.
GLprimer1
23-mer  /  35% GC
1 binding site
5565 .. 5587  =  23 annealed bases
Tm  =  50°C
RVprimer4
20-mer  /  65% GC
1 binding site
2796 .. 2815  =  20 annealed bases
Tm  =  61°C
GLprimer2
23-mer  /  39% GC
1 binding site
77 .. 99  =  23 annealed bases
Tm  =  57°C
luciferase
76 .. 1728  =  1653 bp
550 amino acids  =  60.7 kDa
Product: firefly luciferase
luciferase
76 .. 1728  =  1653 bp
550 amino acids  =  60.7 kDa
Product: firefly luciferase
AmpR
3815 .. 4675  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3815 .. 4606  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3815 .. 4675  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4607 .. 4675  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3815 .. 4675  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3056 .. 3644  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
3056 .. 3644  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4807 .. 5262  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4807 .. 5262  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 poly(A) signal
2609 .. 2743  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2609 .. 2743  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5452 .. 5573  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5452 .. 5573  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4676 .. 4780  =  105 bp
AmpR promoter
4676 .. 4780  =  105 bp
small t intron
1969 .. 2034  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
1969 .. 2034  =  66 bp
simian virus 40 (SV40) small t antigen intron
MCS
1 .. 52  =  52 bp
multiple cloning site
MCS
1 .. 52  =  52 bp
multiple cloning site
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