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Plasmid Files

pBI-CMV5

Mammalian vector for expressing a protein of interest together with secreted Metridia luciferase.

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pBI-CMV5 Sequence and MappBI-CMV5.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SacI (3717) Eco53kI (3715) EcoRI (3680) AleI (3651) EcoNI (3632) BclI * (3511) XcmI (3476) SmaI (3366) TspMI - XmaI (3364) Bpu10I (3345) BsrGI (3310) AarI - BfuAI - BspMI (3307) PluTI (3249) SfoI (3247) NarI (3246) KasI (3245) ApaI (3015) PspOMI (3011) BglII (3005) Bsu36I (2999) PstI (2996) XbaI (2986) SspI (2676) EarI (2667) XmnI (2471) ScaI (2352) PvuI (2242) FspI (2094) NmeAIII (2020) StuI (3730) PaeR7I - PspXI - XhoI (1) NdeI (240) BsaAI - SnaBI (346) BamHI (602) MluI (620) NheI (626) BmtI (630) EagI - NotI (633) BspDI - ClaI (641) HindIII (646) SalI (652) AccI (653) EcoRV (660) PciI (979) NspI (983) DrdI (1087) BseYI (1283) PspFI (1287) AlwNI (1395) AhdI (1872) pBI-CMV5 3759 bp
SacI  (3717)
1 site
G A G C T C C T C G A G
Eco53kI  (3715)
1 site
G A G C T C C T C G A G
EcoRI  (3680)
1 site
G A A T T C C T T A A G
AleI  (3651)
1 site
C A C N N N N G T G G T G N N N N C A C
EcoNI  (3632)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BclI  (3511)
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.
XcmI  (3476)
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.
SmaI  (3366)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3364)
1 site
C C C G G G G G G C C C
XmaI  (3364)
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.
Bpu10I  (3345)
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.
BsrGI  (3310)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
AarI  (3307)
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.
BfuAI  (3307)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (3307)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PluTI  (3249)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (3247)
1 site
G G C G C C C C G C G G
NarI  (3246)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (3245)
1 site
G G C G C C C C G C G G
ApaI  (3015)
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  (3011)
1 site
G G G C C C C C C G G G
BglII  (3005)
1 site
A G A T C T T C T A G A
Bsu36I  (2999)
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.
PstI  (2996)
1 site
C T G C A G G A C G T C
XbaI  (2986)
1 site
T C T A G A A G A T C T
SspI  (2676)
1 site
A A T A T T T T A T A A
EarI  (2667)
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.
XmnI  (2471)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2352)
1 site
A G T A C T T C A T G A
PvuI  (2242)
1 site
C G A T C G G C T A G C
FspI  (2094)
1 site
T G C G C A A C G C G T
NmeAIII  (2020)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
StuI  (3730)
1 site
A G G C C T T C C G G A
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1)
1 site
C T C G A G G A G C T C
NdeI  (240)
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.
BsaAI  (346)
1 site
Y A C G T R R T G C A Y
SnaBI  (346)
1 site
T A C G T A A T G C A T
BamHI  (602)
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.
MluI  (620)
1 site
A C G C G T T G C G C A
NheI  (626)
1 site
G C T A G C C G A T C G
BmtI  (630)
1 site
G C T A G C C G A T C G
EagI  (633)
1 site
C G G C C G G C C G G C
NotI  (633)
1 site
G C G G C C G C C G C C G G C G
BspDI  (641)
1 site
A T C G A T T A G C T A
ClaI  (641)
1 site
A T C G A T T A G C T A
HindIII  (646)
1 site
A A G C T T T T C G A A
SalI  (652)
1 site
G T C G A C C A G C T G
AccI  (653)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
EcoRV  (660)
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.
PciI  (979)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (983)
1 site
R C A T G Y Y G T A C R
DrdI  (1087)
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.
BseYI  (1283)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
PspFI  (1287)
1 site
C C C A G C G G G T C G
AlwNI  (1395)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (1872)
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.
AmpR
1799 .. 2659  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1799 .. 2590  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1799 .. 2659  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   2591 .. 2659  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1799 .. 2659  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
MetLuc
3017 .. 3676  =  660 bp
219 amino acids  =  23.9 kDa
   Segment 2:  
   3017 .. 3625  =  609 bp
   202 amino acids  =  22.0 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
3017 .. 3676  =  660 bp
219 amino acids  =  23.9 kDa
   Segment 1:  signal sequence  
   3626 .. 3676  =  51 bp
   17 amino acids  =  1.9 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
3017 .. 3676  =  660 bp
219 amino acids  =  23.9 kDa
2 segments
Product: secreted Metridia luciferase
human codon-optimized
ori
1040 .. 1628  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1040 .. 1628  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
67 .. 370  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
67 .. 370  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
371 .. 574  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
371 .. 574  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
AmpR promoter
2660 .. 2764  =  105 bp
AmpR promoter
2660 .. 2764  =  105 bp
SV40 poly(A) signal
777 .. 858  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
777 .. 858  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2797 .. 2878  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2797 .. 2878  =  82 bp
SV40 polyadenylation signal
MCS
602 .. 663  =  62 bp
multiple cloning site
MCS
602 .. 663  =  62 bp
multiple cloning site
minimal CMV promoter
3714 .. 3748  =  35 bp
human cytomegalovirus (CMV) immediate early
promoter
minimal CMV promoter
3714 .. 3748  =  35 bp
human cytomegalovirus (CMV) immediate early
promoter
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