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Plasmid Files

pBABE-Hygro

Retroviral expression vector derived from the Moloney murine leukemia virus. Contains a hygromycin resistance marker.

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pBABE-Hygro Sequence and MappBABE-Hygro.dna
Map and Sequence File   
Sequence Author:  Cell Biolabs
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 SspI (5447) XmnI (5242) FspI (4865) NotI (4522) AflIII - PciI (3743) BspQI - SapI (3627) BbsI (3491) BmtI (2873) NheI (2869) AarI (8) PfoI (490) splice donor mutation SpeI (616) Bsu36I (737) BstEII (1035) BsrGI (1230) NgoMIV (1348) NaeI (1350) BamHI (1355) BsaAI - SnaBI (1374) EcoRI (1379) SalI (1397) AccI (1398) SfiI (1680) StuI (1726) AvrII (1727) HindIII (1743) PshAI (1797) AsiSI (2134) RsrII (2178) NdeI (2220) BtgZI (2442) SacII (2550) BspDI - ClaI (2795) pBABE-Hygro 5558 bp
SspI  (5447)
1 site
A A T A T T T T A T A A
XmnI  (5242)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (4865)
1 site
T G C G C A A C G C G T
NotI  (4522)
1 site
G C G G C C G C C G C C G G C G
AflIII  (3743)
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  (3743)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3627)
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  (3627)
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.
BbsI  (3491)
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  (2873)
1 site
G C T A G C C G A T C G
NheI  (2869)
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
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.
NgoMIV  (1348)
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.
NaeI  (1350)
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.
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.
BsaAI  (1374)
1 site
Y A C G T R R T G C A Y
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
PshAI  (1797)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AsiSI  (2134)
1 site
G C G A T C G C C G C T A G C G
RsrII  (2178)
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.
NdeI  (2220)
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.
BtgZI  (2442)
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.
SacII  (2550)
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.
BspDI  (2795)
1 site
A T C G A T T A G C T A
ClaI  (2795)
1 site
A T C G A T T A G C T A
HygR
1770 .. 2792  =  1023 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
1770 .. 2792  =  1023 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
4570 .. 5430  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4570 .. 5361  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4570 .. 5430  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5362 .. 5430  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4570 .. 5430  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
LTR
2839 .. 3432  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
LTR
2839 .. 3432  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
ori
3804 .. 4392  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3804 .. 4392  =  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
5431 .. 5535  =  105 bp
AmpR promoter
5431 .. 5535  =  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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