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pCMV-Gaussia Luc

Control vector for constitutive high-level expression of secreted Gaussia luciferase under control of the CMV promoter.

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pCMV-Gaussia Luc.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Pierce)
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AarI (5453) EcoRV (5159) ApaI (4819) PspOMI (4815) AflII (4806) DrdI (4473) MauBI (3877) ScaI (3360) FspI (3102) AfeI (2930) BstZ17I (2765) SpeI (1) NdeI (236) SnaBI (342) Eco53kI (568) SacI (570) BamHI (634) BsaBI * (677) BbsI (723) NotI (1206) BseRI (1771) AvrII (1775) TspMI - XmaI (1796) SmaI (1798) BclI * (1826) PflFI - Tth111I (1946) BsiWI (1960) RsrII (2020) BstEII (2038) EarI (2063) BsmBI (2296) DraIII (2475) NgoMIV (2576) NaeI (2578) BsmI (2713) pCMV-Gaussia Luc 5483 bp
AarI  (5453)
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  (5159)
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  (4819)
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  (4815)
1 site
G G G C C C C C C G G G
AflII  (4806)
1 site
C T T A A G G A A T T C
DrdI  (4473)
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  (3877)
1 site
C G C G C G C G G C G C G C G C
ScaI  (3360)
1 site
A G T A C T T C A T G A
FspI  (3102)
1 site
T G C G C A A C G C G T
AfeI  (2930)
1 site
A G C G C T T C G C G A
BstZ17I  (2765)
1 site
G T A T A C C A T A T G
SpeI  (1)
1 site
A C T A G T T G A T C A
NdeI  (236)
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.
SnaBI  (342)
1 site
T A C G T A A T G C A T
Eco53kI  (568)
1 site
G A G C T C C T C G A G
SacI  (570)
1 site
G A G C T C C T C G A G
BamHI  (634)
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  (677)
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.
BbsI  (723)
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.
NotI  (1206)
1 site
G C G G C C G C C G C C G G C G
BseRI  (1771)
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.
AvrII  (1775)
1 site
C C T A G G G G A T C C
TspMI  (1796)
1 site
C C C G G G G G G C C C
XmaI  (1796)
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  (1798)
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  (1826)
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  (1946)
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  (1946)
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  (1960)
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  (2020)
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  (2038)
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.
EarI  (2063)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI recognition sequence.
Sticky ends from different EarI sites may not be compatible.
BsmBI  (2296)
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  (2475)
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  (2576)
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  (2578)
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.
BsmI  (2713)
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.
AmpR
2807 .. 3667  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2807 .. 3598  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2807 .. 3667  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3599 .. 3667  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2807 .. 3667  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
1904 .. 2503  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1904 .. 2503  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
3931 .. 4519  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3931 .. 4519  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hGLuc
647 .. 1204  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
hGLuc
647 .. 1204  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
SV40 promoter
1461 .. 1790  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1461 .. 1790  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
63 .. 366  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
63 .. 366  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
367 .. 570  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
367 .. 570  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
2633 .. 2754  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2633 .. 2754  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
1227 .. 1338  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1227 .. 1338  =  112 bp
bovine growth hormone polyadenylation signal
pause site
5392 .. 5483  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5392 .. 5483  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
EM7 promoter
1838 .. 1885  =  48 bp
synthetic bacterial promoter
EM7 promoter
1838 .. 1885  =  48 bp
synthetic bacterial promoter
T7 promoter
615 .. 633  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
615 .. 633  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
5261 .. 5277  =  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
5261 .. 5277  =  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
1641 .. 1776  =  136 bp
SV40 origin of replication
SV40 ori
1641 .. 1776  =  136 bp
SV40 origin of replication
ORF:  526 .. 774  =  249 bp
ORF:  82 amino acids  =  9.7 kDa
ORF:  647 .. 1204  =  558 bp
ORF:  185 amino acids  =  19.9 kDa
ORF:  1904 .. 2503  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  2937 .. 3203  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  2064 .. 2390  =  327 bp
ORF:  108 amino acids  =  12.0 kDa
ORF:  2807 .. 3667  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  577 .. 915  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
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