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pBI-CMV2

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

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pBI-CMV2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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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