pBApo-CMV

Simple mammalian expression vector with a CMV promoter.

Sequence Author: TaKaRa

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EcoRV (3444) ClaI (3440) BspQI - SapI (3195) AflIII - PciI (3078) PspFI (2778) BseYI (2774) AlwNI (2669) AhdI (2190) BpmI (2121) BsrFI (2105) NmeAIII (2043) TsoI (1792) ApoI - EcoRI (1) BtgZI (333) BsaAI - SnaBI (339) BtgI - NcoI - StyI (359) CMV promoter BamHI (581) XbaI (587) SalI (593) AccI (594) HincII (595) PstI - SbfI (603) BfuAI - BspMI (606) SphI (609) HindIII (611) PasI (741) EcoRV (793) ClaI (795) KasI (963) NarI (964) SfoI (965) PluTI (967) BstAPI (1020) PfoI (1151) EcoO109I (1210) SspI (1385) XmnI (1590) ScaI (1709) pBApo-CMV 3447 bp
EcoRV  (3444)
2 sites
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.
ClaI  (3440)
2 sites
A T C G A T T A G C T A
BspQI  (3195)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3195)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
AflIII  (3078)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (3078)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2778)
1 site
C C C A G C G G G T C G
BseYI  (2774)
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.
AlwNI  (2669)
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  (2190)
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.
BpmI  (2121)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BsrFI  (2105)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
NmeAIII  (2043)
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).
TsoI  (1792)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
ApoI  (1)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (1)
1 site
G A A T T C C T T A A G
BtgZI  (333)
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.
BsaAI  (339)
1 site
Y A C G T R R T G C A Y
SnaBI  (339)
1 site
T A C G T A A T G C A T
BtgI  (359)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (359)
1 site
C C A T G G G G T A C C
StyI  (359)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BamHI  (581)
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.
XbaI  (587)
1 site
T C T A G A A G A T C T
SalI  (593)
1 site
G T C G A C C A G C T G
AccI  (594)
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.
HincII  (595)
1 site
G T Y R A C C A R Y T G
PstI  (603)
1 site
C T G C A G G A C G T C
SbfI  (603)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (606)
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  (606)
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.
SphI  (609)
1 site
G C A T G C C G T A C G
HindIII  (611)
1 site
A A G C T T T T C G A A
PasI  (741)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
EcoRV  (793)
2 sites
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.
ClaI  (795)
2 sites
A T C G A T T A G C T A
KasI  (963)
1 site
G G C G C C C C G C G G
NarI  (964)
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.
SfoI  (965)
1 site
G G C G C C C C G C G G
PluTI  (967)
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.
BstAPI  (1020)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PfoI  (1151)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
EcoO109I  (1210)
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.
SspI  (1385)
1 site
A A T A T T T T A T A A
XmnI  (1590)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1709)
1 site
A G T A C T T C A T G A
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1403 .. 1471  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1472 .. 2263  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2434 .. 3022  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2434 .. 3022  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
60 .. 363  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
60 .. 363  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
364 .. 567  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
364 .. 567  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AmpR promoter
1298 .. 1402  =  105 bp
AmpR promoter
1298 .. 1402  =  105 bp
HSV TK poly(A) signal
674 .. 721  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
674 .. 721  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
M13 fwd
808 .. 824  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
808 .. 824  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1403 .. 2263  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  625 .. 1017  =  393 bp
ORF:  130 amino acids  =  14.9 kDa
ORF:  1867 .. 2133  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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