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Plasmid Files

pTK-Gaussia-Dura Luc

Control vector for constitutive low-level expression of secreted Gaussia-Dura luciferase under control of the HSV TK promoter.

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pTK-Gaussia-Dura Luc Sequence and MappTK-Gaussia-Dura Luc.dna
Map and Sequence File   
Sequence Author:  Thermo Scientific (Pierce)
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 SpeI (1) AarI (5596) EcoRV (5302) ApaI (4962) PspOMI (4958) DrdI (4616) MauBI (4020) ScaI (3503) FspI (3245) AfeI (3073) BsmI (2856) PaeR7I - XhoI (7) HindIII (20) BspQI - SapI (155) PpuMI (337) BsaI (404) PvuII (549) BstBI (671) MluI (731) BamHI (778) BsaBI * (820) NotI (1349) BseRI (1914) BclI * (1969) PflFI - Tth111I (2089) BsiWI (2103) RsrII (2163) BstEII (2181) BsmBI (2439) DraIII (2618) NgoMIV (2719) NaeI (2721) pTK-Gaussia-Dura Luc 5626 bp
SpeI  (1)
1 site
A C T A G T T G A T C A
AarI  (5596)
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.
EcoRV  (5302)
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.
ApaI  (4962)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4958)
1 site
G G G C C C C C C G G G
DrdI  (4616)
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.
MauBI  (4020)
1 site
C G C G C G C G G C G C G C G C
ScaI  (3503)
1 site
A G T A C T T C A T G A
FspI  (3245)
1 site
T G C G C A A C G C G T
AfeI  (3073)
1 site
A G C G C T T C G C G A
BsmI  (2856)
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.
PaeR7I  (7)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (7)
1 site
C T C G A G G A G C T C
HindIII  (20)
1 site
A A G C T T T T C G A A
BspQI  (155)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (155)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PpuMI  (337)
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.
BsaI  (404)
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.
PvuII  (549)
1 site
C A G C T G G T C G A C
BstBI  (671)
1 site
T T C G A A A A G C T T
MluI  (731)
1 site
A C G C G T T G C G C A
BamHI  (778)
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.
BsaBI  (820)
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.
NotI  (1349)
1 site
G C G G C C G C C G C C G G C G
BseRI  (1914)
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.
BclI  (1969)
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.
PflFI  (2089)
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  (2089)
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.
BsiWI  (2103)
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.
RsrII  (2163)
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.
BstEII  (2181)
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.
BsmBI  (2439)
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.
DraIII  (2618)
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.
NgoMIV  (2719)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (2721)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
AmpR
2950 .. 3810  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2950 .. 3741  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2950 .. 3810  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3742 .. 3810  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2950 .. 3810  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
HSV TK promoter
26 .. 777  =  752 bp
herpes simplex virus thymidine kinase promoter
HSV TK promoter
26 .. 777  =  752 bp
herpes simplex virus thymidine kinase promoter
PuroR
2047 .. 2646  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2047 .. 2646  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4074 .. 4662  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4074 .. 4662  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
Gaussia-Dura Luc
790 .. 1347  =  558 bp
185 amino acids  =  19.9 kDa
Product: enhanced variant of secreted Gaussia
luciferase
human codon-optimized
Gaussia-Dura Luc
790 .. 1347  =  558 bp
185 amino acids  =  19.9 kDa
Product: enhanced variant of secreted Gaussia
luciferase
human codon-optimized
SV40 promoter
1604 .. 1933  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1604 .. 1933  =  330 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2776 .. 2897  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2776 .. 2897  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
1370 .. 1481  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1370 .. 1481  =  112 bp
bovine growth hormone polyadenylation signal
pause site
5535 .. 5626  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
5535 .. 5626  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
EM7 promoter
1981 .. 2028  =  48 bp
synthetic bacterial promoter
EM7 promoter
1981 .. 2028  =  48 bp
synthetic bacterial promoter
lac operator
5404 .. 5420  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
5404 .. 5420  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
SV40 ori
1784 .. 1919  =  136 bp
SV40 origin of replication
SV40 ori
1784 .. 1919  =  136 bp
SV40 origin of replication
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