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pBApo-CMV Neo

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

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pBApo-CMV Neo.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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ClaI (4546) BsaBI * (4537) MfeI (4445) PsiI (4416) RsrII (4150) NaeI (4136) NgoMIV (4134) BssHII (4031) BanII (3999) PflFI - Tth111I (3752) MscI (3716) EagI (3540) PpuMI (3491) BmeT110I (3430) AbsI - AvaI - BsoBI - PaeR7I - PspXI - XhoI (3429) AvrII (3407) StuI (3406) SfiI (3360) SexAI * (3174) AflIII - PciI (3078) AlwNI (2669) AhdI (2190) EcoRV (4550) EcoRI (1) SnaBI (339) BamHI (581) XbaI (587) SalI (593) AccI (594) SbfI (603) HindIII (611) PasI (741) EcoRV (793) ClaI (795) SspI (1385) XmnI (1590) ScaI (1709) AseI (2015) BpmI (2121) pBApo-CMV Neo 4553 bp
ClaI  (4546)
2 sites		 A T C G A T T A G C T A
BsaBI  (4537)
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  (4445)
1 site		 C A A T T G G T T A A C
PsiI  (4416)
1 site		 T T A T A A A A T A T T
RsrII  (4150)
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.
NaeI  (4136)
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  (4134)
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.
BssHII  (4031)
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.
BanII  (3999)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
PflFI  (3752)
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  (3752)
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  (3716)
1 site		 T G G C C A A C C G G T
EagI  (3540)
1 site		 C G G C C G G C C G G C
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.
BmeT110I  (3430)
1 site		 C Y C G R G G R G C Y C
AbsI  (3429)
1 site		 C C T C G A G G G G A G C T C C
AvaI  (3429)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (3429)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°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
StuI  (3406)
1 site		 A G G C C T T C C G G A
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.
SexAI  (3174)
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.
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  (4550)
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
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
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.
SbfI  (603)
1 site		 C C T G C A G G G G A C G T C C
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
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
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.
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
NeoR/KanR
3506 .. 4300  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3506 .. 4300  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
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
4302 .. 4436  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4302 .. 4436  =  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 .. 4300  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3678 .. 4064  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3815 .. 4453  =  639 bp
ORF:  212 amino acids  =  23.6 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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