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Plasmid Files

pBACgus-2cp

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

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pBACgus-2cp Sequence and MappBACgus-2cp.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 DraIII (7268) ScaI (6596) NmeAIII (6264) BpmI (6186) PspFI (5531) BseYI (5527) PmeI (5103) SgrAI (4815) AgeI (4575) BmgBI (4231) AatII (4008) ZraI (4006) SwaI (3817) SphI (3650) BlpI (3640) BstXI (315) NsiI (332) BbvCI - Bpu10I (564) baculovirus recombination region (lef2/ORF603) BstBI (1882) MscI (2356) PshAI (2404) BglII (3310) NcoI (3413) ATG 6xHis SacII (3443) thrombin site PflMI (3504) NheI (3514) BmtI (3518) enterokinase site BseRI - TspMI - XmaI (3546) SmaI - SrfI (3548) StuI * (3566) BamHI (3570) EcoRI (3576) Eco53kI (3584) SacI (3586) HindIII (3589) EagI - NotI (3596) PaeR7I - PspXI - XhoI (3604) 6xHis AvrII (3631) pBACgus-2cp 7560 bp
DraIII  (7268)
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  (6596)
1 site
A G T A C T T C A T G A
NmeAIII  (6264)
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  (6186)
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  (5531)
1 site
C C C A G C G G G T C G
BseYI  (5527)
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  (5103)
1 site
G T T T A A A C C A A A T T T G
SgrAI  (4815)
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  (4575)
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.
BmgBI  (4231)
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  (4008)
1 site
G A C G T C C T G C A G
ZraI  (4006)
1 site
G A C G T C C T G C A G
SwaI  (3817)
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  (3650)
1 site
G C A T G C C G T A C G
BlpI  (3640)
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

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.
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  (3413)
1 site
C C A T G G G G T A C C
SacII  (3443)
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  (3504)
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  (3514)
1 site
G C T A G C C G A T C G
BmtI  (3518)
1 site
G C T A G C C G A T C G
BseRI  (3546)
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  (3546)
1 site
C C C G G G G G G C C C
XmaI  (3546)
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  (3548)
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  (3548)
1 site
G C C C G G G C C G G G C C C G
StuI  (3566)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BamHI  (3570)
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  (3576)
1 site
G A A T T C C T T A A G
Eco53kI  (3584)
1 site
G A G C T C C T C G A G
SacI  (3586)
1 site
G A G C T C C T C G A G
HindIII  (3589)
1 site
A A G C T T T T C G A A
EagI  (3596)
1 site
C G G C C G G C C G G C
NotI  (3596)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (3604)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3604)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3604)
1 site
C T C G A G G A G C T C
AvrII  (3631)
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)
3664 .. 5021  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
3664 .. 5021  =  1358 bp
contains part of ORF1629
AmpR
6043 .. 6903  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6043 .. 6834  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6043 .. 6903  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6835 .. 6903  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6043 .. 6903  =  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
5284 .. 5872  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5284 .. 5872  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
7035 .. 7490  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
7035 .. 7490  =  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
AmpR promoter
6904 .. 7008  =  105 bp
AmpR promoter
6904 .. 7008  =  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
3546 .. 3609  =  64 bp
multiple cloning site
MCS
3546 .. 3609  =  64 bp
multiple cloning site
S-Tag
3469 .. 3513  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
S-Tag
3469 .. 3513  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
6xHis
3421 .. 3438  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3421 .. 3438  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
3448 .. 3465  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
3448 .. 3465  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
6xHis
3610 .. 3627  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3610 .. 3627  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
enterokinase site
3529 .. 3543  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
3529 .. 3543  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
ATG
3415 .. 3417  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3415 .. 3417  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF1629
3680 .. 5021  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
3680 .. 5021  =  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
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