pAcAB3

Baculovirus transfer plasmid for high-level expression of up to three proteins simultaneously in insect cells.

Sequence Author: AB Vector

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BspQI - SapI (9857) AlwNI (9331) BpmI (8783) ScaI (8371) NdeI (7679) EagI (7227) FspAI (6550) SgrDI (6397) SgrAI (6118) SnaBI (5030) PasI (485) BtgI (867) SacII (870) BstEII (922) StyI (1500) PaeR7I - PspXI - XhoI (1900) SphI (2134) BclI * (2231) 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) BamHI (4960) pAcAB3 10,096 bp
BspQI  (9857)
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  (9857)
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  (9331)
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  (8783)
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  (8371)
1 site
A G T A C T T C A T G A
NdeI  (7679)
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  (7227)
1 site
C G G C C G G C C G G C
FspAI  (6550)
1 site
R T G C G C A Y Y A C G C G T R
SgrDI  (6397)
1 site
C G T C G A C G G C A G C T G C
SgrAI  (6118)
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  (5030)
1 site
T A C G T A A T G C A T
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
BclI  (2231)
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.
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

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.
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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.
BamHI  (4960)
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.
baculovirus recombination region (ORF1629)
4967 .. 6324  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4967 .. 6324  =  1358 bp
contains part of ORF1629
AmpR
8065 .. 8925  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8065 .. 8133  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8065 .. 8925  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8134 .. 8925  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8065 .. 8925  =  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
9096 .. 9684  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9096 .. 9684  =  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
4839 .. 4948  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
4839 .. 4948  =  110 bp
baculovirus promoter for expression in insect cells
AmpR promoter
7960 .. 8064  =  105 bp
AmpR promoter
7960 .. 8064  =  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
ORF1629
4983 .. 6324  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4983 .. 6324  =  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
ORF:  8065 .. 8925  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  968 .. 1240  =  273 bp
ORF:  90 amino acids  =  9.7 kDa
ORF:  2351 .. 2680  =  330 bp
ORF:  109 amino acids  =  12.4 kDa
ORF:  6617 .. 7207  =  591 bp
ORF:  196 amino acids  =  22.9 kDa
ORF:  7415 .. 7720  =  306 bp
ORF:  101 amino acids  =  11.6 kDa
ORF:  75 .. 581  =  507 bp
ORF:  168 amino acids  =  19.3 kDa
ORF:  2661 .. 3293  =  633 bp
ORF:  210 amino acids  =  23.9 kDa
ORF:  3920 .. 4345  =  426 bp
ORF:  141 amino acids  =  16.6 kDa
ORF:  613 .. 1599  =  987 bp
ORF:  328 amino acids  =  37.8 kDa
ORF:  1867 .. 2322  =  456 bp
ORF:  151 amino acids  =  17.6 kDa
ORF:  3331 .. 3936  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  6280 .. 6618  =  339 bp
ORF:  112 amino acids  =  13.3 kDa
ORF:  7360 .. 7680  =  321 bp
ORF:  106 amino acids  =  12.4 kDa
ORF:  3375 .. 3605  =  231 bp
ORF:  76 amino acids  =  9.3 kDa
ORF:  4983 .. 6236  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
ORF:  8529 .. 8795  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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