Resources
Plasmid Files

pBacPAK8

Baculovirus vector for strong constitutive expression of proteins in insect cells. For an alternative MCS, use pBacPAK9.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pBacPAK8 Sequence and MappBacPAK8.dna
Map and Sequence File   
Sequence Author:  Clontech
Download Free Trial Get SnapGene Viewer

 BspQI - SapI (5475) PciI (5358) PspFI (5058) BseYI (5054) AlwNI (4949) BmrI (4430) BpmI (4401) BglI (4352) NmeAIII (4323) PvuI (4101) ScaI (3989) EcoO109I (3490) PfoI (3431) DraIII (3074) BsmI (83) BstXI (313) NsiI (330) MluI (545) BbvCI - Bpu10I (562) BseRI (678) baculovirus recombination region (lef2/ORF603) EcoRV (1157) Bac1 Primer (1181 .. 1200) BsaBI (1182) BamHI (1252) PstI - SbfI (1262) StuI (1264) PaeR7I - PspXI - XhoI (1266) BstBI (1273) XbaI (1278) BglII (1284) Acc65I (1290) KpnI (1294) Eco53kI (1298) SacI (1300) EcoRI (1302) TspMI - XmaI (1307) SmaI (1309) EagI - NotI (1313) PacI (1324) Bac2 Primer (1355 .. 1374) SnaBI (1400) SwaI (1490) HindIII (1763) BmgBI (1904) AgeI (2248) AleI (2479) SgrAI (2488) pBacPAK8 5538 bp
BspQI  (5475)
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  (5475)
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.
PciI  (5358)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (5058)
1 site
C C C A G C G G G T C G
BseYI  (5054)
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.
AlwNI  (4949)
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.
BmrI  (4430)
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.
BpmI  (4401)
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.
BglI  (4352)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
NmeAIII  (4323)
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).
PvuI  (4101)
1 site
C G A T C G G C T A G C
ScaI  (3989)
1 site
A G T A C T T C A T G A
EcoO109I  (3490)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PfoI  (3431)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
DraIII  (3074)
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.
BsmI  (83)
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.
BstXI  (313)
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  (330)
1 site
A T G C A T T A C G T A
MluI  (545)
1 site
A C G C G T T G C G C A
BbvCI  (562)
1 site
C C T C A G C G G A G T C G
Bpu10I  (562)
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.
BseRI  (678)
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  (1157)
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.
BsaBI  (1182)
1 site
G A T N N N N A T C C T A N N N N T A G
BamHI  (1252)
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.
PstI  (1262)
1 site
C T G C A G G A C G T C
SbfI  (1262)
1 site
C C T G C A G G G G A C G T C C
StuI  (1264)
1 site
A G G C C T T C C G G A
PaeR7I  (1266)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1266)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1266)
1 site
C T C G A G G A G C T C
BstBI  (1273)
1 site
T T C G A A A A G C T T
XbaI  (1278)
1 site
T C T A G A A G A T C T
BglII  (1284)
1 site
A G A T C T T C T A G A
Acc65I  (1290)
1 site
G G T A C C C C A T G G
KpnI  (1294)
1 site
G G T A C C C C A T G G
Eco53kI  (1298)
1 site
G A G C T C C T C G A G
SacI  (1300)
1 site
G A G C T C C T C G A G
EcoRI  (1302)
1 site
G A A T T C C T T A A G
TspMI  (1307)
1 site
C C C G G G G G G C C C
XmaI  (1307)
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  (1309)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EagI  (1313)
1 site
C G G C C G G C C G G C
NotI  (1313)
1 site
G C G G C C G C C G C C G G C G
PacI  (1324)
1 site
T T A A T T A A A A T T A A T T
SnaBI  (1400)
1 site
T A C G T A A T G C A T
SwaI  (1490)
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.
HindIII  (1763)
1 site
A A G C T T T T C G A A
BmgBI  (1904)
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.
AgeI  (2248)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AleI  (2479)
1 site
C A C N N N N G T G G T G N N N N C A C
SgrAI  (2488)
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.
Bac1 Primer
20-mer  /  30% GC
1 binding site
1181 .. 1200  =  20 annealed bases
Tm  =  49°C
Bac2 Primer
20-mer  /  50% GC
1 binding site
1355 .. 1374  =  20 annealed bases
Tm  =  59°C
ORF1629
1353 .. 2694  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
1353 .. 2694  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
AmpR
3683 .. 4543  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3683 .. 3751  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3683 .. 4543  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3752 .. 4543  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3683 .. 4543  =  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)
327 .. 1154  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
327 .. 1154  =  828 bp
contains ORF603 and part of lef2
ori
4714 .. 5302  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4714 .. 5302  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2841 .. 3296  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2841 .. 3296  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
3578 .. 3682  =  105 bp
AmpR promoter
3578 .. 3682  =  105 bp
polyhedrin promoter
1158 .. 1249  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
1158 .. 1249  =  92 bp
promoter for the baculovirus polyhedrin gene
MCS
1252 .. 1327  =  76 bp
multiple cloning site
MCS
1252 .. 1327  =  76 bp
multiple cloning site
ORF603
489 .. 1094  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
489 .. 1094  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
327 .. 451  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
327 .. 451  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
stop codons
1321 .. 1331  =  11 bp
stop codons in all three reading frames
stop codons
1321 .. 1331  =  11 bp
stop codons in all three reading frames
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter