pcDNA4 HisMax-TOPO (linearized)

Linearized mammalian vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products and expression of N-terminally 6xHis-tagged proteins.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTA and GC Cloning VectorspcDNA4 HisMax-TOPO (linearized)
SgrDI (4088) SspI (3971) ScaI (3647) PvuI (3537) FspI (3389) AhdI (3167) PciI (2277) BspQI - SapI (2161) BstZ17I (1898) BsmI (1846) FseI (1538) BmgBI (1396) SgrAI (1376) BglII (4102) MfeI (4251) Bpu10I (4270) NruI (4298) MluI (4318) SpeI (4339) NdeI (4574) SnaBI (4680) Eco53kI (4906) SacI (4908) AflII (4998) HindIII (5001) BlpI (5004) SrfI (5051) ATG 6xHis T7 tag (gene 10 leader) Xpress™ tag End (5274) Start (1) BamHI (16) EcoRI (31) PstI (40) EcoRV (43) NotI (58) PaeR7I - PspXI - XhoI (64) XbaI (70) PspOMI (76) ApaI (80) BbsI (296) StuI (1132) MscI (1265) MauBI (1298) pcDNA™4/HisMax-TOPO® 5273 bp
SgrDI  (4088)
1 site		 C G T C G A C G G C A G C T G C
SspI  (3971)
1 site		 A A T A T T T T A T A A
ScaI  (3647)
1 site		 A G T A C T T C A T G A
PvuI  (3537)
1 site		 C G A T C G G C T A G C
FspI  (3389)
1 site		 T G C G C A A C G C G T
AhdI  (3167)
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.
PciI  (2277)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2161)
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  (2161)
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.
BstZ17I  (1898)
1 site		 G T A T A C C A T A T G
BsmI  (1846)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
FseI  (1538)
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.
BmgBI  (1396)
1 site		 C A C G T C G T G C A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
SgrAI  (1376)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BglII  (4102)
1 site		 A G A T C T T C T A G A
MfeI  (4251)
1 site		 C A A T T G G T T A A C
Bpu10I  (4270)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
NruI  (4298)
1 site		 T C G C G A A G C G C T
MluI  (4318)
1 site		 A C G C G T T G C G C A
SpeI  (4339)
1 site		 A C T A G T T G A T C A
NdeI  (4574)
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.
SnaBI  (4680)
1 site		 T A C G T A A T G C A T
Eco53kI  (4906)
1 site		 G A G C T C C T C G A G
SacI  (4908)
1 site		 G A G C T C C T C G A G
AflII  (4998)
1 site		 C T T A A G G A A T T C
HindIII  (5001)
1 site		 A A G C T T T T C G A A
BlpI  (5004)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
SrfI  (5051)
1 site		 G C C C G G G C C G G G C C C G
End  (5274)
0 sites
Start  (1)
0 sites
BamHI  (16)
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.
EcoRI  (31)
1 site		 G A A T T C C T T A A G
PstI  (40)
1 site		 C T G C