pBApo-CMV Pur

Simple mammalian expression vector with a CMV promoter and a puromycin resistance marker.

Sequence Author: TaKaRa

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ClaI (4351) BsaBI * (4342) MfeI (4250) PsiI (4221) DraIII (4077) BssHII (3961) EagI (3954) MscI * (3881) SacII (3720) BstEII (3640) RsrII (3622) BsiWI (3562) SmaI (3557) TspMI - XmaI (3555) PflFI - Tth111I (3548) PpuMI (3491) AbsI - PaeR7I - PspXI - XhoI (3429) AvrII (3407) SfiI (3360) AflIII - PciI (3078) AlwNI (2669) AhdI (2190) EcoRV (4355) EcoRI (1) BtgZI (333) BsaAI - SnaBI (339) BamHI (581) XbaI (587) PstI - SbfI (603) BfuAI - BspMI (606) HindIII (611) PasI (741) EcoRV (793) ClaI (795) M13 fwd PvuII (894) SspI (1385) XmnI (1590) ScaI (1709) AseI (2015) pBApo-CMV Pur 4358 bp
ClaI  (4351)
2 sites
A T C G A T T A G C T A
BsaBI  (4342)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MfeI  (4250)
1 site
C A A T T G G T T A A C
PsiI  (4221)
1 site
T T A T A A A A T A T T
DraIII  (4077)
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.
BssHII  (3961)
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.
EagI  (3954)
1 site
C G G C C G G C C G G C
MscI  (3881)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
SacII  (3720)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BstEII  (3640)
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.
RsrII  (3622)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsiWI  (3562)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
SmaI  (3557)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3555)
1 site
C C C G G G G G G C C C
XmaI  (3555)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
PflFI  (3548)
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  (3548)
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.
PpuMI  (3491)
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.
AbsI  (3429)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (3429)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3429)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3429)
1 site
C T C G A G G A G C T C
AvrII  (3407)
1 site
C C T A G G G G A T C C
SfiI  (3360)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
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.
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.
EcoRV  (4355)
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.
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
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
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.
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
PvuII  (894)
1 site
C A G C T G G T C G A C
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
AseI  (2015)
1 site
A T T A A T T A A T T 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
PuroR
3506 .. 4105  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3506 .. 4105  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
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
SV40 promoter
3093 .. 3422  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3093 .. 3422  =  330 bp
SV40 enhancer and early promoter
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
SV40 poly(A) signal
4107 .. 4241  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4107 .. 4241  =  135 bp
SV40 polyadenylation signal
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)
MCS
581 .. 616  =  36 bp
multiple cloning site
MCS
581 .. 616  =  36 bp
multiple cloning site
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
SV40 ori
3273 .. 3408  =  136 bp
SV40 origin of replication
SV40 ori
3273 .. 3408  =  136 bp
SV40 origin of replication
ORF:  1403 .. 2263  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3080 .. 3370  =  291 bp
ORF:  96 amino acids  =  10.9 kDa
ORF:  3506 .. 4105  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  3488 .. 4258  =  771 bp
ORF:  256 amino acids  =  26.6 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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