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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 Sequence and MappMCS-Cypridina Luc.dna
Map and Sequence File   
Sequence Author:  Thermo Scientific (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) MCS 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) EM7 promoter 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

The sticky ends produced by AflII are hard to ligate.
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
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