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Plasmid Files

pAcAB4

Baculovirus transfer vector for expressing up to four proteins simultaneously, for use with the BD BaculoGold™ system.

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pAcAB4 Sequence and MappAcAB4.dna
Map and Sequence File   
Sequence Author:  BD Biosciences
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 BspQI - SapI (9991) AlwNI (9465) BpmI (8917) ScaI (8505) NdeI (7813) EagI (7361) FspAI (6684) SgrDI (6531) SgrAI (6252) SnaBI (5164) BamHI (5094) polyhedrin promoter PasI (485) BtgI (867) SacII (870) BstEII (922) StyI (1500) PaeR7I - PspXI - XhoI (1900) SphI (2134) BbvCI - Bpu10I (3404) baculovirus recombination region (lef2/ORF603) NgoMIV (3769) NaeI (3771) BlpI (4440) BglII (4454) PpuMI (4568) StuI (4706) XbaI (4709) polyhedrin promoter TspMI - XmaI (4955) SmaI (4957) SpeI (4961) pAcAB4 10,230 bp
BspQI  (9991)
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  (9991)
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.
AlwNI  (9465)
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.
BpmI  (8917)
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.
ScaI  (8505)
1 site
A G T A C T T C A T G A
NdeI  (7813)
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.
EagI  (7361)
1 site
C G G C C G G C C G G C
FspAI  (6684)
1 site
R T G C G C A Y Y A C G C G T R
SgrDI  (6531)
1 site
C G T C G A C G G C A G C T G C
SgrAI  (6252)
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.
SnaBI  (5164)
1 site
T A C G T A A T G C A T
BamHI  (5094)
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.
PasI  (485)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
BtgI  (867)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (870)
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.
BstEII  (922)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
StyI  (1500)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PaeR7I  (1900)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1900)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1900)
1 site
C T C G A G G A G C T C
SphI  (2134)
1 site
G C A T G C C G T A C G
BbvCI  (3404)
1 site
C C T C A G C G G A G T C G
Bpu10I  (3404)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
NgoMIV  (3769)
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  (3771)
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.
BlpI  (4440)
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.
BglII  (4454)
1 site
A G A T C T T C T A G A
PpuMI  (4568)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
StuI  (4706)
1 site
A G G C C T T C C G G A
XbaI  (4709)
1 site
T C T A G A A G A T C T
TspMI  (4955)
1 site
C C C G G G G G G C C C
XmaI  (4955)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (4957)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SpeI  (4961)
1 site
A C T A G T T G A T C A
baculovirus recombination region (ORF1629)
5101 .. 6458  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
5101 .. 6458  =  1358 bp
contains part of ORF1629
AmpR
8199 .. 9059  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8199 .. 8267  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8199 .. 9059  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8268 .. 9059  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8199 .. 9059  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
baculovirus recombination region (lef2/ORF603)
3169 .. 3996  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
3169 .. 3996  =  828 bp
contains ORF603 and part of lef2
ori
9230 .. 9818  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
9230 .. 9818  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 poly(A) signal
4012 .. 4133  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4012 .. 4133  =  122 bp
SV40 polyadenylation signal
p10 promoter
4462 .. 4571  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
4462 .. 4571  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
4838 .. 4947  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
4838 .. 4947  =  110 bp
baculovirus promoter for expression in insect cells
AmpR promoter
8094 .. 8198  =  105 bp
AmpR promoter
8094 .. 8198  =  105 bp
polyhedrin promoter
4722 .. 4813  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4722 .. 4813  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
5000 .. 5091  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
5000 .. 5091  =  92 bp
promoter for the baculovirus polyhedrin gene
ORF1629
5117 .. 6458  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
5117 .. 6458  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
3331 .. 3936  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
3331 .. 3936  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
3169 .. 3293  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
3169 .. 3293  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
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