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Plasmid Files

pTK-neo

Mammalian vector encoding a neomycin resistance gene, to select for stable integration of a cotransfected expression plasmid.

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pTK-neo Sequence and MappTK-neo.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 BbsI (2868) ApaLI (2548) Bsu36I (2056) BstAPI (2045) PfoI (1719) RsrII (1460) BssHII (231) TspMI - XmaI (272) SmaI (274) SacII (289) PpuMI (309) AccI (332) BstZ17I (333) ApoI - EcoRI (639) AflIII - MluI (703) StuI (752) BspDI * - ClaI * (784) BsaBI * (803) EagI (850) BmrI (893) FspI (1046) PflFI - Tth111I (1062) NmeAIII (1168) BsrDI (1177) SphI (1349) pTK-neo 2872 bp
BbsI  (2868)
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.
ApaLI  (2548)
1 site
G T G C A C C A C G T G
Bsu36I  (2056)
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.
BstAPI  (2045)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PfoI  (1719)
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.
RsrII  (1460)
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  (231)
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.
TspMI  (272)
1 site
C C C G G G G G G C C C
XmaI  (272)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (274)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SacII  (289)
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.
PpuMI  (309)
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.
AccI  (332)
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.
BstZ17I  (333)
1 site
G T A T A C C A T A T G
ApoI  (639)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (639)
1 site
G A A T T C C T T A A G
AflIII  (703)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (703)
1 site
A C G C G T T G C G C A
StuI  (752)
1 site
A G G C C T T C C G G A
BspDI  (784)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (784)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BsaBI  (803)
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.
EagI  (850)
1 site
C G G C C G G C C G G C
BmrI  (893)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
FspI  (1046)
1 site
T G C G C A A C G C G T
PflFI  (1062)
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  (1062)
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.
NmeAIII  (1168)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrDI  (1177)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
SphI  (1349)
1 site
G C A T G C C G T A C G
NeoR/KanR
816 .. 1610  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
816 .. 1610  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
HSV TK promoter
2870 .. 749  =  752 bp
herpes simplex virus thymidine kinase promoter
HSV TK promoter
2870 .. 749  =  752 bp
herpes simplex virus thymidine kinase promoter
ori
2218 .. 2806  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2218 .. 2806  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
HSV TK poly(A) signal
1842 .. 1889  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
1842 .. 1889  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
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