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Plasmid Files

pBABE-Neo

Retroviral expression vector derived from the Moloney murine leukemia virus. Contains a neomycin (G418) resistance marker.

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pBABE-Neo Sequence and MappBABE-Neo.dna
Map and Sequence File   
Sequence Author:  Cell Biolabs
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 SspI (5219) XmnI (5014) ScaI (4895) PvuI (4785) NotI (4294) AflIII - PciI (3515) BbsI (3263) BmtI (2645) AarI (8) PfoI (490) SpeI (616) ZraI (699) AatII (701) Bsu36I (737) BstEII (1035) BsrGI (1230) BamHI (1355) SnaBI (1374) EcoRI (1379) SalI (1397) AccI (1398) SfiI (1680) StuI (1726) AvrII (1727) HindIII (1743) RsrII (2414) BspDI * - ClaI * (2567) NheI (2641) pBABE-Neo 5330 bp
SspI  (5219)
1 site
A A T A T T T T A T A A
XmnI  (5014)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (4895)
1 site
A G T A C T T C A T G A
PvuI  (4785)
1 site
C G A T C G G C T A G C
NotI  (4294)
1 site
G C G G C C G C C G C C G G C G
AflIII  (3515)
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  (3515)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BbsI  (3263)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BmtI  (2645)
1 site
G C T A G C C G A T C G
AarI  (8)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
PfoI  (490)
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.
SpeI  (616)
1 site
A C T A G T T G A T C A
ZraI  (699)
1 site
G A C G T C C T G C A G
AatII  (701)
1 site
G A C G T C C T G C A G
Bsu36I  (737)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BstEII  (1035)
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.
BsrGI  (1230)
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.
BamHI  (1355)
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.
SnaBI  (1374)
1 site
T A C G T A A T G C A T
EcoRI  (1379)
1 site
G A A T T C C T T A A G
SalI  (1397)
1 site
G T C G A C C A G C T G
AccI  (1398)
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.
SfiI  (1680)
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.
StuI  (1726)
1 site
A G G C C T T C C G G A
AvrII  (1727)
1 site
C C T A G G G G A T C C
HindIII  (1743)
1 site
A A G C T T T T C G A A
RsrII  (2414)
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.
BspDI  (2567)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2567)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
NheI  (2641)
1 site
G C T A G C C G A T C G
AmpR
4342 .. 5202  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4342 .. 5133  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4342 .. 5202  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5134 .. 5202  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4342 .. 5202  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
1770 .. 2564  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1770 .. 2564  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
LTR
2611 .. 3204  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
LTR
2611 .. 3204  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
ori
3576 .. 4164  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3576 .. 4164  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
LTR (truncated)
8 .. 479  =  472 bp
long terminal repeat from Moloney murine leukemia
virus
LTR (truncated)
8 .. 479  =  472 bp
long terminal repeat from Moloney murine leukemia
virus
gag (truncated)
934 .. 1350  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
934 .. 1350  =  417 bp
truncated MMLV gag gene lacking the start codon
SV40 promoter
1413 .. 1742  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1413 .. 1742  =  330 bp
SV40 enhancer and early promoter
MMLV Ψ
542 .. 741  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
542 .. 741  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
AmpR promoter
5203 .. 5307  =  105 bp
AmpR promoter
5203 .. 5307  =  105 bp
MCS
1355 .. 1402  =  48 bp
multiple cloning site
MCS
1355 .. 1402  =  48 bp
multiple cloning site
splice donor mutation
541 .. 541  =  1 bp
T-to-C mutation that blocks RNA splicing
splice donor mutation
541 .. 541  =  1 bp
T-to-C mutation that blocks RNA splicing
SV40 ori
1593 .. 1728  =  136 bp
SV40 origin of replication
SV40 ori
1593 .. 1728  =  136 bp
SV40 origin of replication
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