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Plasmid Files

pMCS-Cypridina Luc

Secreted Cypridina luciferase vector for cloning a transcriptional regulatory element.

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pMCS-Cypridina Luc.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Pierce)
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SpeI (1) AarI (5959) AflII (5312) DrdI (4979) MauBI (4383) NruI (4310) PvuI (3756) FspI (3608) AfeI (3436) SalI (3276) BstZ17I (3271) EcoRI (7) Acc65I (13) KpnI (17) Eco53kI (21) SacI (23) PaeR7I - XhoI (25) HindIII (31) BamHI (37) BbsI (58) Bpu10I (211) BstXI (402) BspDI - ClaI (473) BsrDI (530) PstI (865) NheI (937) BmtI (941) AleI (997) BlpI (1003) Bsu36I (1310) KflI - PpuMI (1365) NotI (1712) NcoI (2188) AvrII (2281) PflFI - Tth111I (2452) BsiWI (2466) RsrII (2526) BstEII (2544) EarI (2569) SacII (2624) BsmBI (2802) DraIII (2981) pMCS-Cypridina Luc 5989 bp
SpeI  (1)
1 site
A C T A G T T G A T C A
AarI  (5959)
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.
AflII  (5312)
1 site
C T T A A G G A A T T C
DrdI  (4979)
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  (4383)
1 site
C G C G C G C G G C G C G C G C
NruI  (4310)
1 site
T C G C G A A G C G C T
PvuI  (3756)
1 site
C G A T C G G C T A G C
FspI  (3608)
1 site
T G C G C A A C G C G T
AfeI  (3436)
1 site
A G C G C T T C G C G A
SalI  (3276)
1 site
G T C G A C C A G C T G
BstZ17I  (3271)
1 site
G T A T A C C A T A T G
EcoRI  (7)
1 site
G A A T T C C T T A A G
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.
BbsI  (58)
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.
Bpu10I  (211)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BstXI  (402)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BspDI  (473)
1 site
A T C G A T T A G C T A
ClaI  (473)
1 site
A T C G A T T A G C T A
BsrDI  (530)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PstI  (865)
1 site
C T G C A G G A C G T C
NheI  (937)
1 site
G C T A G C C G A T C G
BmtI  (941)
1 site
G C T A G C C G A T C G
AleI  (997)
1 site
C A C N N N N G T G G T G N N N N C A C
BlpI  (1003)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
Bsu36I  (1310)
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.
KflI  (1365)
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  (1365)
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.
NotI  (1712)
1 site
G C G G C C G C C G C C G G C G
NcoI  (2188)
1 site
C C A T G G G G T A C C
AvrII  (2281)
1 site
C C T A G G G G A T C C
PflFI  (2452)
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  (2452)
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  (2466)
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  (2526)
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  (2544)
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  (2569)
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.
SacII  (2624)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BsmBI  (2802)
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  (2981)
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.
hCluc
49 .. 1710  =  1662 bp
553 amino acids  =  61.5 kDa
Product: secreted Cypridina luciferase
human codon-optimized
hCluc
49 .. 1710  =  1662 bp
553 amino acids  =  61.5 kDa
Product: secreted Cypridina luciferase
human codon-optimized
AmpR
3313 .. 4173  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3313 .. 4104  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3313 .. 4173  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4105 .. 4173  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3313 .. 4173  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
2410 .. 3009  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2410 .. 3009  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4437 .. 5025  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4437 .. 5025  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
1967 .. 2296  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1967 .. 2296  =  330 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
3139 .. 3260  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3139 .. 3260  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
1733 .. 1844  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1733 .. 1844  =  112 bp
bovine growth hormone polyadenylation signal
pause site
5898 .. 5989  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5898 .. 5989  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
EM7 promoter
2344 .. 2391  =  48 bp
synthetic bacterial promoter
EM7 promoter
2344 .. 2391  =  48 bp
synthetic bacterial promoter
MCS
1 .. 42  =  42 bp
multiple cloning site
MCS
1 .. 42  =  42 bp
multiple cloning site
lac operator
5767 .. 5783  =  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
5767 .. 5783  =  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
2147 .. 2282  =  136 bp
SV40 origin of replication
SV40 ori
2147 .. 2282  =  136 bp
SV40 origin of replication
ORF:  49 .. 1710  =  1662 bp
ORF:  553 amino acids  =  61.5 kDa
ORF:  2410 .. 3009  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  3443 .. 3709  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  171 .. 788  =  618 bp
ORF:  205 amino acids  =  24.3 kDa
ORF:  969 .. 1313  =  345 bp
ORF:  114 amino acids  =  13.1 kDa
ORF:  2570 .. 2896  =  327 bp
ORF:  108 amino acids  =  12.0 kDa
ORF:  3313 .. 4173  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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