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pBApo-EF1α Neo

Simple mammalian expression vector with an EF-1α promoter and a neomycin resistance marker.

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pBApo-EF1(alpha) Neo.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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EcoRV (5164) ClaI (5160) BsaBI * (5151) PsiI (5030) RsrII (4764) BssHII (4645) PflFI - Tth111I (4366) MscI (4330) EagI (4154) PpuMI (4105) AbsI - PspXI (4043) AvrII (4021) SfiI (3974) SexAI * (3788) AlwNI (3283) AhdI (2804) BpmI (2735) EcoRI (1) AgeI (89) BstXI (322) AflII (394) BglII (581) PspOMI (628) ApaI (632) FseI (719) Eco53kI (841) SacI (843) BbvCI - Bpu10I (917) DraIII (1051) BamHI (1195) XbaI (1201) SalI (1207) AccI (1208) SbfI (1217) HindIII (1225) EcoRV (1407) ClaI (1409) NdeI (1630) ZraI (1883) AatII (1885) SspI (1999) ScaI (2323) AseI (2629) pBApo-EF1 α Neo 5167 bp
EcoRV  (5164)
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  (5160)
2 sites
A T C G A T T A G C T A
BsaBI  (5151)
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.
PsiI  (5030)
1 site
T T A T A A A A T A T T
RsrII  (4764)
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.
BssHII  (4645)
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.
PflFI  (4366)
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  (4366)
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.
MscI  (4330)
1 site
T G G C C A A C C G G T
EagI  (4154)
1 site
C G G C C G G C C G G C
PpuMI  (4105)
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  (4043)
1 site
C C T C G A G G G G A G C T C C
PspXI  (4043)
1 site
V C T C G A G B B G A G C T C V
AvrII  (4021)
1 site
C C T A G G G G A T C C
SfiI  (3974)
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.
SexAI  (3788)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AlwNI  (3283)
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  (2804)
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  (2735)
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.
EcoRI  (1)
1 site
G A A T T C C T T A A G
AgeI  (89)
1 site
A C C G G T T G G C C A
BstXI  (322)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
AflII  (394)
1 site
C T T A A G G A A T T C
BglII  (581)
1 site
A G A T C T T C T A G A
PspOMI  (628)
1 site
G G G C C C C C C G G G
ApaI  (632)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
FseI  (719)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
Eco53kI  (841)
1 site
G A G C T C C T C G A G
SacI  (843)
1 site
G A G C T C C T C G A G
BbvCI  (917)
1 site
C C T C A G C G G A G T C G
Bpu10I  (917)
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.
DraIII  (1051)
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.
BamHI  (1195)
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  (1201)
1 site
T C T A G A A G A T C T
SalI  (1207)
1 site
G T C G A C C A G C T G
AccI  (1208)
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.
SbfI  (1217)
1 site
C C T G C A G G G G A C G T C C
HindIII  (1225)
1 site
A A G C T T T T C G A A
EcoRV  (1407)
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  (1409)
2 sites
A T C G A T T A G C T A
NdeI  (1630)
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.
ZraI  (1883)
1 site
G A C G T C C T G C A G
AatII  (1885)
1 site
G A C G T C C T G C A G
SspI  (1999)
1 site
A A T A T T T T A T A A
ScaI  (2323)
1 site
A G T A C T T C A T G A
AseI  (2629)
1 site
A T T A A T T A A T T A
EF-1α promoter
12 .. 1193  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
12 .. 1193  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
2017 .. 2877  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2017 .. 2085  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2017 .. 2877  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2086 .. 2877  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2017 .. 2877  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
4120 .. 4914  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
4120 .. 4914  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3048 .. 3636  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3048 .. 3636  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
3707 .. 4036  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3707 .. 4036  =  330 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
4916 .. 5050  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4916 .. 5050  =  135 bp
SV40 polyadenylation signal
AmpR promoter
1912 .. 2016  =  105 bp
AmpR promoter
1912 .. 2016  =  105 bp
HSV TK poly(A) signal
1288 .. 1335  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
1288 .. 1335  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
M13 fwd
1422 .. 1438  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
1422 .. 1438  =  17 bp
common sequencing primer, one of multiple similar variants
EF-1α intron A
242 .. 1184  =  943 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
242 .. 1184  =  943 bp
intron upstream of the start codon of human EF-1α
SV40 ori
3887 .. 4022  =  136 bp
SV40 origin of replication
SV40 ori
3887 .. 4022  =  136 bp
SV40 origin of replication
ORF:  571 .. 1008  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  2017 .. 2877  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3694 .. 3984  =  291 bp
ORF:  96 amino acids  =  10.9 kDa
ORF:  4120 .. 4914  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  650 .. 883  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  4292 .. 4678  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  286 .. 933  =  648 bp
ORF:  215 amino acids  =  23.7 kDa
ORF:  4429 .. 5067  =  639 bp
ORF:  212 amino acids  =  23.6 kDa
ORF:  1239 .. 1631  =  393 bp
ORF:  130 amino acids  =  14.9 kDa
ORF:  2481 .. 2747  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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