pFastBac Dual

Dual-promoter vector for co-expression of two proteins in insect cells using the Bac-to-Bac® baculovirus system.

Sequence Author: Thermo Fisher (Invitrogen)

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AvrII (4978) HpaI (4827) MfeI (4814) HindIII (4699) PstI (4690) XbaI (4678) BstBI (4673) NotI (4664) SpeI (4656) SacI (4654) Eco53kI (4652) SalI - SgrDI (4644) StuI (4638) EcoRI (4628) BssHII (4621) RsrII (4613) BamHI (4606) BstZ17I (4458) PacI (4384) BbsI (4325) SmaI (4321) TspMI - XmaI (4319) PaeR7I - XhoI (4311) MCS NcoI (4305) BmtI (4302) NheI (4298) PvuII (4295) NsiI (4291) SphI (4284) KpnI (4278) Acc65I (4274) HSV TK poly(A) signal SnaBI (3982) BsrGI (3510) PflFI - Tth111I (3328) BsmBI - Esp3I (3282) BseRI (3061) EcoRV (2925) SacII (2868) BstXI (2817) NgoMIV (227) NaeI (229) BsaHI (936) PvuI (1107) FspI (1253) BpmI (1407) AlwNI (1955) BseYI (2060) PspFI (2064) BspQI - SapI (2481) MscI (2809) pFastBac™ Dual 5238 bp
AvrII  (4978)
1 site
C C T A G G G G A T C C
HpaI  (4827)
1 site
G T T A A C C A A T T G
MfeI  (4814)
1 site
C A A T T G G T T A A C
HindIII  (4699)
1 site
A A G C T T T T C G A A
PstI  (4690)
1 site
C T G C A G G A C G T C
XbaI  (4678)
1 site
T C T A G A A G A T C T
BstBI  (4673)
1 site
T T C G A A A A G C T T
NotI  (4664)
1 site
G C G G C C G C C G C C G G C G
SpeI  (4656)
1 site
A C T A G T T G A T C A
SacI  (4654)
1 site
G A G C T C C T C G A G
Eco53kI  (4652)
1 site
G A G C T C C T C G A G
SalI  (4644)
1 site
G T C G A C C A G C T G
SgrDI  (4644)
1 site
C G T C G A C G G C A G C T G C
StuI  (4638)
1 site
A G G C C T T C C G G A
EcoRI  (4628)
1 site
G A A T T C C T T A A G
BssHII  (4621)
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.
RsrII  (4613)
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.
BamHI  (4606)
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.
BstZ17I  (4458)
1 site
G T A T A C C A T A T G
PacI  (4384)
1 site
T T A A T T A A A A T T A A T T
BbsI  (4325)
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.
SmaI  (4321)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (4319)
1 site
C C C G G G G G G C C C
XmaI  (4319)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
PaeR7I  (4311)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (4311)
1 site
C T C G A G G A G C T C
NcoI  (4305)
1 site
C C A T G G G G T A C C
BmtI  (4302)
1 site
G C T A G C C G A T C G
NheI  (4298)
1 site
G C T A G C C G A T C G
PvuII  (4295)
1 site
C A G C T G G T C G A C
NsiI  (4291)
1 site
A T G C A T T A C G T A
SphI  (4284)
1 site
G C A T G C C G T A C G
KpnI  (4278)
1 site
G G T A C C C C A T G G
Acc65I  (4274)
1 site
G G T A C C C C A T G G
SnaBI  (3982)
1 site
T A C G T A A T G C A T
BsrGI  (3510)
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  (3328)
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  (3328)
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  (3282)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (3282)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BseRI  (3061)
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  (2925)
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  (2868)
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  (2817)
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.
NgoMIV  (227)
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  (229)
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.
BsaHI  (936)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
PvuI  (1107)
1 site
C G A T C G G C T A G C
FspI  (1253)
1 site
T G C G C A A C G C G T
BpmI  (1407)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AlwNI  (1955)
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.
BseYI  (2060)
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.
PspFI  (2064)
1 site
C C C A G C G G G T C G
BspQI  (2481)
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  (2481)
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.