pBACgus-3

Baculovirus transfer plasmid with the polyhedrin promoter, encoding β-glucuronidase plus an N-terminal signal sequence-6xHis-thrombin-S-Tag-enterokinase cassette.

Sequence Author: MilliporeSigma (Novagen)

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DraIII (7331) ScaI (6659) NmeAIII (6327) BpmI (6249) PspFI (5594) BseYI (5590) PmeI (5166) SgrAI (4878) AgeI (4638) BmgBI (4294) AatII (4071) ZraI (4069) SwaI (3880) SphI (3713) BlpI (3703) BstXI (315) NsiI (332) BbvCI - Bpu10I (564) baculovirus recombination region (lef2/ORF603) BstBI (1882) MscI (2356) PshAI (2404) BglII (3310) GP64 signal sequence NcoI (3476) 6xHis SacII (3506) thrombin site PflMI (3567) NheI (3577) BmtI (3581) enterokinase site BseRI - TspMI - XmaI (3609) SmaI - SrfI (3611) StuI * (3629) BamHI (3633) EcoRI (3639) Eco53kI (3647) SacI (3649) HindIII (3652) EagI - NotI (3659) PaeR7I - PspXI - XhoI (3667) 6xHis AvrII (3694) pBACgus-3 7623 bp
DraIII  (7331)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
ScaI  (6659)
1 site
A G T A C T T C A T G A
NmeAIII  (6327)
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).
BpmI  (6249)
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.
PspFI  (5594)
1 site
C C C A G C G G G T C G
BseYI  (5590)
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.
PmeI  (5166)
1 site
G T T T A A A C C A A A T T T G
SgrAI  (4878)
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.
AgeI  (4638)
1 site
A C C G G T T G G C C A
BmgBI  (4294)
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.
AatII  (4071)
1 site
G A C G T C C T G C A G
ZraI  (4069)
1 site
G A C G T C C T G C A G
SwaI  (3880)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
SphI  (3713)
1 site
G C A T G C C G T A C G
BlpI  (3703)
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.
BstXI  (315)
1 site
C C A N N N N N N T G G G G T N N