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pFastBac™/HBM-TOPO® (linearized)

Linearized baculovirus expression and secretion vector, for TOPO® cloning of blunt PCR products and addition of a cleavable C-terminal 6xHis tag.

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pFastBac HBM-TOPO (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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SnaBI (4587) BbsI (4463) BsrGI (4116) PflFI - Tth111I (3934) BsmBI (3888) BseRI (3667) EcoRV (3531) SacII (3474) BstXI (3423) MscI (3415) BspQI - SapI (3085) PspFI (2668) BseYI (2664) AlwNI (2559) BamHI (4738) AgeI (4744) BspDI * - ClaI * (4815) End (4824) Start (0) PaeR7I - PspXI - XhoI (49) SphI (59) Acc65I (61) KpnI (65) HindIII (67) MfeI (182) HpaI (195) BclI * (331) AvrII (346) NgoMIV (833) NaeI (835) BanII (867) BsaHI (1540) PvuI (1711) FspI (1857) BpmI (2011) pFastBac™/HBM-TOPO® 4824 bp
SnaBI  (4587)
1 site
T A C G T A A T G C A T
BbsI  (4463)
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.
BsrGI  (4116)
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.
PflFI  (3934)
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  (3934)
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.
BsmBI  (3888)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BseRI  (3667)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
EcoRV  (3531)
1 site
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.
SacII  (3474)
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.
BstXI  (3423)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
MscI  (3415)
1 site
T G G C C A A C C G G T
BspQI  (3085)
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  (3085)
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.
PspFI  (2668)
1 site
C C C A G C G G G T C G
BseYI  (2664)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2559)
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.
BamHI  (4738)
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.
AgeI  (4744)
1 site
A C C G G T T G G C C A
BspDI  (4815)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4815)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
End  (4824)
0 sites
Start  (0)
0 sites
PaeR7I  (49)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (49)
1 site
V C T C G A G B B G A G C T C V
XhoI  (49)
1 site
C T C G A G G A G C T C
SphI  (59)
1 site
G C A T G C C G T A C G
Acc65I  (61)
1 site
G G T A C C C C A T G G
KpnI  (65)
1 site
G G T A C C C C A T G G
HindIII  (67)
1 site
A A G C T T T T C G A A
MfeI  (182)
1 site
C A A T T G G T T A A C
HpaI  (195)
1 site
G T T A A C C A A T T G
BclI  (331)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
AvrII  (346)
1 site
C C T A G G G G A T C C
NgoMIV  (833)
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  (835)
1 site