pSV40-CLuc

Control vector with a strong constitutive SV40 promoter for expressing secreted Cypridina luciferase in mammalian cells.

Sequence Author: New England Biolabs

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Acc65I (1) poly(A) signal EagI - NotI (5089) DraIII (4743) XmnI (4190) BsaHI (4128) AseI (3763) BsaI (3652) AhdI (3591) BspQI - SapI (2582) KpnI (5) NheI (18) BmtI (22) PstI (27) Eco53kI (30) SacI (32) EcoRI (34) EcoRV (42) HindIII (46) SfiI (185) StuI (231) AvrII (232) AvaI - BsoBI - PaeR7I - XhoI (247) SpeI (272) XcmI (689) BstEII (800) PmlI (1187) BmgBI (1210) EcoO109I - KflI - PpuMI (1299) BclI * (1787) PflFI - Tth111I (1821) Bsu36I (1851) MluI (1953) XbaI (1961) HpaI (2097) MfeI (2106) SalI (2448) AfeI (2574) pSV40-CLuc 5256 bp
Acc65I  (1)
1 site
G G T A C C C C A T G G
EagI  (5089)
1 site
C G G C C G G C C G G C
NotI  (5089)
1 site
G C G G C C G C C G C C G G C G
DraIII  (4743)
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  (4190)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (4128)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
AseI  (3763)
1 site
A T T A A T T A A T T A
BsaI  (3652)
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  (3591)
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.
BspQI  (2582)
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  (2582)
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.
KpnI  (5)
1 site
G G T A C C C C A T G G
NheI  (18)
1 site
G C T A G C C G A T C G
BmtI  (22)
1 site
G C T A G C C G A T C G
PstI  (27)
1 site
C T G C A G G A C G T C
Eco53kI  (30)
1 site
G A G C T C C T C G A G
SacI  (32)
1 site
G A G C T C C T C G A G
EcoRI  (34)
1 site
G A A T T C C T T A A G
EcoRV  (42)
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.
HindIII  (46)
1 site
A A G C T T T T C G A A
SfiI  (185)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
StuI  (231)
1 site
A G G C C T T C C G G A
AvrII  (232)
1 site
C C T A G G G G A T C C
AvaI  (247)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (247)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (247)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (247)
1 site
C T C G A G G A G C T C
SpeI  (272)
1 site
A C T A G T T G A T C A
XcmI  (689)
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.
BstEII  (800)
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.
PmlI  (1187)
1 site
C A C G T G G T G C A C
BmgBI  (1210)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
EcoO109I  (1299)
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.
KflI  (1299)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1299)
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.
BclI  (1787)
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  (1821)
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  (1821)
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.
Bsu36I  (1851)
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.
MluI  (1953)
1 site
A C G C G T T G C G C A
XbaI  (1961)
1 site
T C T A G A A G A T C T
HpaI  (2097)
1 site
G T T A A C C A A T T G
MfeI  (2106)
1 site
C A A T T G G T T A A C
SalI  (2448)
1 site
G T C G A C C A G C T G
AfeI  (2574)
1 site
A G C G C T T C G C G A
CLuc
291 .. 1952  =  1662 bp
553 amino acids  =  61.5 kDa
Product: secreted Cypridina luciferase
contains 30 codon substitutions for efficient translation in mammalian cells
CLuc
291 .. 1952  =  1662 bp
553 amino acids  =  61.5 kDa
Product: secreted Cypridina luciferase
contains 30 codon substitutions for efficient translation in mammalian cells
AmpR
3518 .. 4378  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3518 .. 4309  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3518 .. 4378  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4310 .. 4378  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3518 .. 4378  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2759 .. 3347  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2759 .. 3347  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4510 .. 4965  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4510 .. 4965  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
52 .. 246  =  195 bp
SV40 early promoter
SV40 promoter
52 .. 246  =  195 bp
SV40 early promoter
SV40 poly(A) signal
1976 .. 2097  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1976 .. 2097  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4379 .. 4483  =  105 bp
AmpR promoter
4379 .. 4483  =  105 bp
pause site
5158 .. 5249  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5158 .. 5249  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
MCS
1 .. 51  =  51 bp
multiple cloning site
MCS
1 .. 51  =  51 bp
multiple cloning site
poly(A) signal
5096 .. 5144  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5096 .. 5144  =  49 bp
synthetic polyadenylation signal
SV40 ori
98 .. 233  =  136 bp
SV40 origin of replication
SV40 ori
98 .. 233  =  136 bp
SV40 origin of replication
ORF:  1045 .. 1356  =  312 bp
ORF:  103 amino acids  =  11.3 kDa
ORF:  291 .. 1952  =  1662 bp
ORF:  553 amino acids  =  61.5 kDa
ORF:  3648 .. 3914  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1660 .. 1884  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  2062 .. 2547  =  486 bp
ORF:  161 amino acids  =  18.0 kDa
ORF:  1548 .. 1916  =  369 bp
ORF:  122 amino acids  =  13.8 kDa
ORF:  3518 .. 4378  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pSV40-CLuc.dna file

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