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Plasmid Files

pBI-CMV2

Bidirectional promoter vector for expressing a gene together with the fluorescent protein AcGFP1.

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pBI-CMV2 Sequence and MappBI-CMV2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SacI (3780) Eco53kI (3778) EcoRI (3743) PluTI (3723) SfoI (3721) NarI (3720) KasI (3719) MscI (3662) BstEII (3548) Bpu10I (3540) PfoI * (3450) BssHII (3405) DraIII (3289) EcoNI (3110) PpuMI (3103) PflMI (3040) ApaI (3015) PspOMI (3011) BglII (3005) PstI (2996) XbaI (2986) SspI (2676) EarI (2667) XmnI (2471) ScaI (2352) PvuI (2242) FspI (2094) BsaI (1933) StuI (3793) 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-CMV2 3822 bp
SacI  (3780)
1 site
G A G C T C C T C G A G
Eco53kI  (3778)
1 site
G A G C T C C T C G A G
EcoRI  (3743)
1 site
G A A T T C C T T A A G
PluTI  (3723)
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  (3721)
1 site
G G C G C C C C G C G G
NarI  (3720)
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  (3719)
1 site
G G C G C C C C G C G G
MscI  (3662)
1 site
T G G C C A A C C G G T
BstEII  (3548)
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.
Bpu10I  (3540)
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.
PfoI  (3450)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BssHII  (3405)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
DraIII  (3289)
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.
EcoNI  (3110)
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.
PpuMI  (3103)
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.
PflMI  (3040)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
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
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
BsaI  (1933)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
StuI  (3793)
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
AcGFP1
3017 .. 3736  =  720 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
AcGFP1
3017 .. 3736  =  720 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
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
3777 .. 3814  =  38 bp
human cytomegalovirus (CMV) immediate early
promoter
minimal CMV promoter
3777 .. 3814  =  38 bp
human cytomegalovirus (CMV) immediate early
promoter
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