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Plasmid Files

pBI-CMV3

Bidirectional promoter vector for expressing a gene together with the fluorescent protein ZsGreen1.

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pBI-CMV3 Sequence and MappBI-CMV3.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SacI (3756) Eco53kI (3754) EcoRI (3719) BclI * (3625) SgrAI (3620) PflFI - Tth111I (3476) NaeI (3445) NgoMIV (3443) PluTI (3408) SfoI (3406) NarI (3405) KasI (3404) BseRI (3353) BmgBI (3220) BfuAI - BspMI (3196) BsrGI (3157) ApaI (3015) EcoO109I (3012) PspOMI (3011) BglII (3005) PstI (2996) XbaI (2986) SspI (2676) EarI (2667) XmnI (2471) ScaI (2352) PvuI (2242) FspI (2094) BsaI (1933) StuI (3769) PaeR7I - PspXI - XhoI (1) NdeI (240) BsaAI - SnaBI (346) BamHI (602) PvuII (616) MluI (620) NheI (626) BmtI (630) EagI - NotI (633) BspDI - ClaI (641) HindIII (646) EcoRV (660) PciI (979) DrdI (1087) BseYI (1283) PspFI (1287) AlwNI (1395) pBI-CMV3 3798 bp
SacI  (3756)
1 site
G A G C T C C T C G A G
Eco53kI  (3754)
1 site
G A G C T C C T C G A G
EcoRI  (3719)
1 site
G A A T T C C T T A A G
BclI  (3625)
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.
SgrAI  (3620)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
PflFI  (3476)
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  (3476)
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.
NaeI  (3445)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (3443)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
PluTI  (3408)
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  (3406)
1 site
G G C G C C C C G C G G
NarI  (3405)
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  (3404)
1 site
G G C G C C C C G C G G
BseRI  (3353)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
BmgBI  (3220)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BfuAI  (3196)
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  (3196)
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.
BsrGI  (3157)
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.
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.
EcoO109I  (3012)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
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  (3769)
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.
PvuII  (616)
1 site
C A G C T G G T C G A C
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
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.
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.
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
ZsGreen1
3017 .. 3712  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
3017 .. 3712  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green 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
3753 .. 3790  =  38 bp
human cytomegalovirus (CMV) immediate early
promoter
minimal CMV promoter
3753 .. 3790  =  38 bp
human cytomegalovirus (CMV) immediate early
promoter
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