pBacPAK8

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

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsInsect Cell VectorspBacPAK8
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
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.
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
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
2 segments
   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
2 segments
   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
ORF:  71 .. 451  =  381 bp
ORF:  126 amino acids  =  14.4 kDa
ORF:  3683 .. 4543  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4147 .. 4413  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  489 .. 1094  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  1353 .. 2606  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
ORF:  533 .. 763  =  231 bp
ORF:  76 amino acids  =  9.3 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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