pBAC-2cp

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

Sequence Author: MilliporeSigma (Novagen)

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BsmI (85) DraIII (5119) ScaI (4447) NmeAIII (4115) BpmI (4037) BmrI (4007) AlwNI (3490) PspFI (3382) BseYI (3378) PciI (3074) PmeI (2954) SgrAI (2666) BstXI (315) NsiI (332) MluI (547) BbvCI - Bpu10I (564) baculovirus recombination region (lef2/ORF603) BglII (1161) BsaBI (1192) NcoI (1264) ATG 6xHis SacII (1294) thrombin site PflMI (1355) NheI (1365) BmtI (1369) enterokinase site BseRI - TspMI - XmaI (1397) SmaI - SrfI (1399) StuI * (1417) BamHI (1421) EcoRI (1427) Eco53kI (1435) SacI (1437) HindIII (1440) EagI - NotI (1447) PaeR7I - PspXI - XhoI (1455) 6xHis AvrII (1482) BlpI (1491) SphI (1501) SnaBI (1578) SwaI (1668) ZraI (1857) AatII (1859) BmgBI (2082) AgeI (2426) AleI (2657) pBAC™-2cp 5411 bp
BsmI  (85)
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.
DraIII  (5119)
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  (4447)
1 site
A G T A C T T C A T G A
NmeAIII  (4115)
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  (4037)
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.
BmrI  (4007)
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.
AlwNI  (3490)
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.
PspFI  (3382)
1 site
C C C A G C G G G T C G
BseYI  (3378)
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.
PciI  (3074)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PmeI  (2954)
1 site
G T T T A A A C C A A A T T T G
SgrAI  (2666)
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.
BstXI  (315)
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.
NsiI  (332)
1 site
A T G C A T T A C G T A
MluI  (547)
1 site
A C G C G T T G C G C A
BbvCI  (564)
1 site
C C T C A G C G G A G T C G
Bpu10I  (564)
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.
BglII  (1161)
1 site
A G A T C T T C T A G A
BsaBI  (1192)
1 site
G A T N N N N A T C C T A N N N N T A G
NcoI  (1264)
1 site
C C A T G G G G T A C C
SacII  (1294)
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.
PflMI  (1355)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
NheI  (1365)
1 site
G C T A G C C G A T C G
BmtI  (1369)
1 site
G C T A G C C G A T C G
BseRI  (1397)
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.
TspMI  (1397)
1 site
C C C G G G G G G C C C
XmaI  (1397)
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.
SmaI  (1399)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (1399)
1 site
G C C C G G G C C G G G C C C G
StuI  (1417)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BamHI  (1421)
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  (1427)
1 site
G A A T T C C T T A A G
Eco53kI  (1435)
1 site
G A G C T C C T C G A G
SacI  (1437)
1 site
G A G C T C C T C G A G
HindIII  (1440)
1 site
A A G C T T