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Plasmid Files

pMCS-Gaussia Luc

Secreted Gaussia luciferase vector for cloning a transcriptional regulatory element.

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pMCS-Gaussia Luc Sequence and MappMCS-Gaussia Luc.dna
Map and Sequence File   
Sequence Author:  Thermo Scientific (Pierce)
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 EcoRI (7) SpeI (1) AarI (4856) EcoRV (4562) ApaI (4222) PspOMI (4218) AflII (4209) DrdI (3876) MauBI (3280) ScaI (2763) FspI (2505) Acc65I (13) KpnI (17) Eco53kI (21) SacI (23) PaeR7I - XhoI (25) HindIII (31) BamHI (37) BsaBI * (80) BbsI (126) NotI (609) AvrII (1178) TspMI - XmaI (1199) SmaI (1201) BclI * (1229) PflFI - Tth111I (1349) BsiWI (1363) RsrII (1423) BstEII (1441) EarI (1466) BsmBI (1699) DraIII (1878) NgoMIV (1979) NaeI (1981) BsmI (2116) BstZ17I (2168) AfeI (2333) pMCS-Gaussia Luc 4886 bp
EcoRI  (7)
1 site
G A A T T C C T T A A G
SpeI  (1)
1 site
A C T A G T T G A T C A
AarI  (4856)
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  (4562)
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  (4222)
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  (4218)
1 site
G G G C C C C C C G G G
AflII  (4209)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
DrdI  (3876)
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  (3280)
1 site
C G C G C G C G G C G C G C G C
ScaI  (2763)
1 site
A G T A C T T C A T G A
FspI  (2505)
1 site
T G C G C A A C G C G T
Acc65I  (13)
1 site
G G T A C C C C A T G G
KpnI  (17)
1 site
G G T A C C C C A T G G
Eco53kI  (21)
1 site
G A G C T C C T C G A G
SacI  (23)
1 site
G A G C T C C T C G A G
PaeR7I  (25)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (25)
1 site
C T C G A G G A G C T C
HindIII  (31)
1 site
A A G C T T T T C G A A
BamHI  (37)
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  (80)
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  (126)
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  (609)
1 site
G C G G C C G C C G C C G G C G
AvrII  (1178)
1 site
C C T A G G G G A T C C
TspMI  (1199)
1 site
C C C G G G G G G C C C
XmaI  (1199)
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  (1201)
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  (1229)
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  (1349)
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  (1349)
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  (1363)
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  (1423)
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  (1441)
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  (1466)
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  (1699)
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  (1878)
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  (1979)
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  (1981)
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  (2116)
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.
BstZ17I  (2168)
1 site
G T A T A C C A T A T G
AfeI  (2333)
1 site
A G C G C T T C G C G A
AmpR
2210 .. 3070  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2210 .. 3001  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2210 .. 3070  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3002 .. 3070  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2210 .. 3070  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
1307 .. 1906  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1307 .. 1906  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
3334 .. 3922  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3334 .. 3922  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hGLuc
50 .. 607  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
hGLuc
50 .. 607  =  558 bp
185 amino acids  =  19.9 kDa
Product: secreted Gaussia luciferase
human codon-optimized
SV40 promoter
864 .. 1193  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
864 .. 1193  =  330 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2036 .. 2157  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2036 .. 2157  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
630 .. 741  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
630 .. 741  =  112 bp
bovine growth hormone polyadenylation signal
pause site
4795 .. 4886  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
4795 .. 4886  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
EM7 promoter
1241 .. 1288  =  48 bp
synthetic bacterial promoter
EM7 promoter
1241 .. 1288  =  48 bp
synthetic bacterial promoter
MCS
1 .. 42  =  42 bp
multiple cloning site
MCS
1 .. 42  =  42 bp
multiple cloning site
lac operator
4664 .. 4680  =  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
4664 .. 4680  =  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
1044 .. 1179  =  136 bp
SV40 origin of replication
SV40 ori
1044 .. 1179  =  136 bp
SV40 origin of replication
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