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Plasmid Files

pBI-CMV4

Bidirectional promoter vector for expressing a gene together with the fluorescent protein DsRed2.

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pBI-CMV4 Sequence and MappBI-CMV4.dna
Map and Sequence File   
Sequence Author:  Clontech
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 Eco53kI (3735) EcoRI (3700) FspAI (3644) BstEII (3547) SbfI (3356) BbsI (3268) SexAI * (3170) BstXI (3154) MscI (3146) BsgI (3115) AleI (3065) ApaI (3015) PspOMI (3011) BglII (3005) Bsu36I (2999) XbaI (2986) SspI (2676) EarI (2667) XmnI (2471) ScaI (2352) PvuI (2242) NmeAIII (2020) SacI (3737) PaeR7I - PspXI - XhoI (1) NdeI (240) BtgZI (340) 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) AlwNI (1395) BsrFI (1952) pBI-CMV4 3779 bp
Eco53kI  (3735)
1 site
G A G C T C C T C G A G
EcoRI  (3700)
1 site
G A A T T C C T T A A G
FspAI  (3644)
1 site
R T G C G C A Y Y A C G C G T R
BstEII  (3547)
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.
SbfI  (3356)
1 site
C C T G C A G G G G A C G T C C
BbsI  (3268)
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.
SexAI  (3170)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BstXI  (3154)
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.
MscI  (3146)
1 site
T G G C C A A C C G G T
BsgI  (3115)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AleI  (3065)
1 site
C A C N N N N G T G G T G N N N N C A C
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.
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
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).
SacI  (3737)
1 site
G A G C T C C T C G A G
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.
BtgZI  (340)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
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.
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.
BsrFI  (1952)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
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
DsRed2
3016 .. 3693  =  678 bp
225 amino acids  =  25.8 kDa
Product: improved tetrameric variant of DsRed
fluorescent protein
mammalian codon-optimized
DsRed2
3016 .. 3693  =  678 bp
225 amino acids  =  25.8 kDa
Product: improved tetrameric variant of DsRed
fluorescent protein
mammalian 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
3734 .. 3771  =  38 bp
human cytomegalovirus (CMV) immediate early
promoter
minimal CMV promoter
3734 .. 3771  =  38 bp
human cytomegalovirus (CMV) immediate early
promoter
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