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Plasmid Files

pXPG

Improved high-copy luciferase plasmid, for expressing cytoplasmic luciferase as a transcriptional reporter.

 
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pXPG.dna
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BamHI BsmBI PflFI - Tth111I BsaAI BstZ17I BstAPI NdeI PciI AlwNI AhdI BsaI BglI NmeAIII FspI PvuI EaeI XmnI AatII ZraI EcoRI SwaI BglII PaeR7I - XhoI Eco53kI SacI Acc65I KpnI - TspMI - XmaI SmaI SalI HindIII NcoI KasI NarI SfoI PluTI BstBI BsrGI BclI * SphI PpuMI AarI SgrAI XbaI PflMI BsgI BsaBI * BspDI * - ClaI * AleI PspOMI ApaI pXPG 6087 bp
BamHI  (1)
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.
BsmBI  (5578)
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.
PflFI  (5477)
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  (5477)
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.
BsaAI  (5471)
1 site
Y A C G T R R T G C A Y
BstZ17I  (5452)
1 site
G T A T A C C A T A T G
BstAPI  (5404)
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.
NdeI  (5400)
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.
PciI  (5221)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (4812)
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  (4333)
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.
BsaI  (4267)
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.
BglI  (4215)
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  (4186)
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  (4110)
1 site
T G C G C A A C G C G T
PvuI  (3964)
1 site
C G A T C G G C T A G C
EaeI  (3940)
1 site
Y G G C C R R C C G G Y
XmnI  (3733)
1 site
G A A N N N N T T C C T T N N N N A A G
AatII  (3414)
1 site
G A C G T C C T G C A G
ZraI  (3412)
1 site
G A C G T C C T G C A G
EcoRI  (3335)
1 site
G A A T T C C T T A A G
SwaI  (3321)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BglII  (13)
1 site
A G A T C T T C T A G A
PaeR7I  (17)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (17)
1 site
C T C G A G G A G C T C
Eco53kI  (22)
1 site
G A G C T C C T C G A G
SacI  (24)
1 site
G A G C T C C T C G A G
Acc65I  (26)
1 site
G G T A C C C C A T G G
KpnI  (30)
1 site
G G T A C C C C A T G G
TspMI  (30)
1 site
C C C G G G G G G C C C
XmaI  (30)
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  (32)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SalI  (35)
1 site
G T C G A C C A G C T G
HindIII  (41)
1 site
A A G C T T T T C G A A
NcoI  (74)
1 site
C C A T G G G G T A C C
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.
BstBI  (245)
1 site
T T C G A A A A G C T T
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.
BclI  (656)
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.
SphI  (739)
1 site
G C A T G C C G T A C G
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.
AarI  (1465)
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.
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.
XbaI  (1730)
1 site
T C T A G A A G A T C T
PflMI  (1888)
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.
BsgI  (2120)
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  (2351)
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  (2583)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2583)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AleI  (2783)
1 site
C A C N N N N G T G G T G N N N N C A C
PspOMI  (2859)
1 site
G G G C C C C C C G G G
ApaI  (2863)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
luciferase
76 .. 1728  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
enhanced luc+ version of the luciferase gene
luciferase
76 .. 1728  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
enhanced luc+ version of the luciferase gene
AmpR
3546 .. 4406  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3546 .. 3614  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3546 .. 4406  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3615 .. 4406  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3546 .. 4406  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4577 .. 5165  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4577 .. 5165  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 poly(A) signal
2450 .. 2584  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2450 .. 2584  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5730 .. 5864  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5730 .. 5864  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5958 .. 5  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5958 .. 5  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3441 .. 3545  =  105 bp
AmpR promoter
3441 .. 3545  =  105 bp
small t intron
1813 .. 1878  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
1813 .. 1878  =  66 bp
simian virus 40 (SV40) small t antigen intron
MCS
1 .. 46  =  46 bp
multiple cloning site
MCS
1 .. 46  =  46 bp
multiple cloning site
ORF:  1957 .. 2430  =  474 bp
ORF:  157 amino acids  =  18.2 kDa
ORF:  5944 .. 1728  =  1872 bp
ORF:  623 amino acids  =  68.8 kDa
ORF:  4604 .. 4831  =  228 bp
ORF:  75 amino acids  =  8.0 kDa
ORF:  2436 .. 2693  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  3546 .. 4406  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6067 .. 213  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  361 .. 588  =  228 bp
ORF:  75 amino acids  =  9.1 kDa
ORF:  2559 .. 3290  =  732 bp
ORF:  243 amino acids  =  27.0 kDa
ORF:  2729 .. 3013  =  285 bp
ORF:  94 amino acids  =  10.5 kDa
ORF:  4010 .. 4276  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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