pBACgus-3

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

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspBACgus-3
DraIII (7331) ScaI (6659) NmeAIII (6327) BpmI (6249) PspFI (5594) BseYI (5590) PmeI (5166) SgrAI (4878) AgeI (4638) BmgBI (4294) AatII (4071) ZraI (4069) SwaI (3880) SphI (3713) BlpI (3703) BstXI (315) NsiI (332) BbvCI - Bpu10I (564) baculovirus recombination region (lef2/ORF603) BstBI (1882) MscI (2356) PshAI (2404) BglII (3310) GP64 signal sequence NcoI (3476) 6xHis SacII (3506) thrombin site PflMI (3567) NheI (3577) BmtI (3581) enterokinase site BseRI - TspMI - XmaI (3609) SmaI - SrfI (3611) StuI * (3629) BamHI (3633) EcoRI (3639) Eco53kI (3647) SacI (3649) HindIII (3652) EagI - NotI (3659) PaeR7I - PspXI - XhoI (3667) 6xHis AvrII (3694) pBACgus-3 7623 bp
DraIII  (7331)
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  (6659)
1 site		 A G T A C T T C A T G A
NmeAIII  (6327)
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  (6249)
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.
PspFI  (5594)
1 site		 C C C A G C G G G T C G
BseYI  (5590)
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.
PmeI  (5166)
1 site		 G T T T A A A C C A A A T T T G
SgrAI  (4878)
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.
AgeI  (4638)
1 site		 A C C G G T T G G C C A
BmgBI  (4294)
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.
AatII  (4071)
1 site		 G A C G T C C T G C A G
ZraI  (4069)
1 site		 G A C G T C C T G C A G
SwaI  (3880)
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.
SphI  (3713)
1 site		 G C A T G C C G T A C G
BlpI  (3703)
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.
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
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.
BstBI  (1882)
1 site		 T T C G A A A A G C T T
MscI  (2356)
1 site		 T G G C C A A C C G G T
PshAI  (2404)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BglII  (3310)
1 site		 A G A T C T T C T A G A
NcoI  (3476)
1 site		 C C A T G G G G T A C C
SacII  (3506)
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  (3567)
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  (3577)
1 site		 G C T A G C C G A T C G
BmtI  (3581)
1 site		 G C T A G C C G A T C G
BseRI  (3609)
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  (3609)
1 site		 C C C G G G G G G C C C
XmaI  (3609)
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  (3611)
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  (3611)
1 site		 G C C C G G G C C G G G C C C G
StuI  (3629)
1 site		 A G G C C T T C C G G A
* Blocked by Dcm methylation.
BamHI  (3633)
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  (3639)
1 site		 G A A T T C C T T A A G
Eco53kI  (3647)
1 site		 G A G C T C C T C G A G
SacI  (3649)
1 site		 G A G C T C C T C G A G
HindIII  (3652)
1 site		 A A G C T T T T C G A A
EagI  (3659)
1 site		 C G G C C G G C C G G C
NotI  (3659)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (3667)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3667)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (3667)
1 site		 C T C G A G G A G C T C
AvrII  (3694)
1 site		 C C T A G G G G A T C C
GUS
1167 .. 2978  =  1812 bp
603 amino acids  =  68.4 kDa
Product: β-glucuronidase
GUS
1167 .. 2978  =  1812 bp
603 amino acids  =  68.4 kDa
Product: β-glucuronidase
baculovirus recombination region (ORF1629)
3727 .. 5084  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
3727 .. 5084  =  1358 bp
contains part of ORF1629
AmpR
6106 .. 6966  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6106 .. 6897  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6106 .. 6966  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6898 .. 6966  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6106 .. 6966  =  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)
329 .. 1156  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
329 .. 1156  =  828 bp
contains ORF603 and part of lef2
ori
5347 .. 5935  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5347 .. 5935  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
7098 .. 7553  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
7098 .. 7553  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
p6.9 promoter
2987 .. 3302  =  316 bp
baculovirus late basic protein promoter
p6.9 promoter
2987 .. 3302  =  316 bp
baculovirus late basic protein promoter
GP64 signal sequence
3415 .. 3474  =  60 bp
20 amino acids  =  2.0 kDa
Product: signal sequence from the baculovirus envelope glycoprotein GP64
GP64 signal sequence
3415 .. 3474  =  60 bp
20 amino acids  =  2.0 kDa
Product: signal sequence from the baculovirus envelope glycoprotein GP64
6xHis
3484 .. 3501  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3484 .. 3501  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
3511 .. 3528  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
3511 .. 3528  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
S-Tag
3532 .. 3576  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
3532 .. 3576  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
enterokinase site
3592 .. 3606  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
3592 .. 3606  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
6xHis
3673 .. 3690  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3673 .. 3690  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
AmpR promoter
6967 .. 7071  =  105 bp
AmpR promoter
6967 .. 7071  =  105 bp
polyhedrin promoter
3317 .. 3408  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
3317 .. 3408  =  92 bp
promoter for the baculovirus polyhedrin gene
MCS
3609 .. 3672  =  64 bp
multiple cloning site
MCS
3609 .. 3672  =  64 bp
multiple cloning site
ORF1629
3743 .. 5084  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
3743 .. 5084  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
491 .. 1096  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
491 .. 1096  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
329 .. 453  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
329 .. 453  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
ORF:  73 .. 453  =  381 bp
ORF:  126 amino acids  =  14.4 kDa
ORF:  3409 .. 3693  =  285 bp
ORF:  94 amino acids  =  9.9 kDa
ORF:  3437 .. 3706  =  270 bp
ORF:  89 amino acids  =  10.4 kDa
ORF:  6236 .. 6502  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1488 .. 1814  =  327 bp
ORF:  108 amino acids  =  12.8 kDa
ORF:  2403 .. 2864  =  462 bp
ORF:  153 amino acids  =  17.5 kDa
ORF:  535 .. 765  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  3019 .. 3345  =  327 bp
ORF:  108 amino acids  =  12.8 kDa
ORF:  6106 .. 6966  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1167 .. 2978  =  1812 bp
ORF:  603 amino acids  =  68.4 kDa
ORF:  491 .. 1096  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  3743 .. 4996  =  1254 bp
ORF:  417 amino acids  =  46.0 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

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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
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Mammalian Expression Vectors
Structural Genomics Vectors

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