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Plasmid Files

pAcUW51

Compact baculovirus transfer vector for expressing two proteins simultaneously, for use with the BD BaculoGold™ system.

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pAcUW51 Sequence and MappAcUW51.dna
Map and Sequence File   
Sequence Author:  BD Biosciences
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 BspQI - SapI (5595) NspI (5482) PciI (5478) DrdI (5376) PspFI (5178) BseYI (5174) AlwNI (5069) BpmI (4521) NmeAIII (4443) ScaI (4109) DraIII (3459) BanII (3385) SgrAI (2740) BbvCI - Bpu10I (173) BsmI (823) XcmI (931) BlpI (1209) EcoRI (1217) BglII (1223) PacI (1270) EcoO109I - PpuMI (1337) NsiI (1358) XbaI (1462) BclI * (1486) BsaBI (1512) BamHI (1582) SnaBI (1652) SwaI (1742) ZraI (1931) AatII (1933) HindIII (2015) BmgBI (2156) AgeI (2500) AleI (2731) pAcUW51 5863 bp
BspQI  (5595)
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  (5595)
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.
NspI  (5482)
1 site
R C A T G Y Y G T A C R
PciI  (5478)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (5376)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PspFI  (5178)
1 site
C C C A G C G G G T C G
BseYI  (5174)
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  (5069)
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  (4521)
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.
NmeAIII  (4443)
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).
ScaI  (4109)
1 site
A G T A C T T C A T G A
DraIII  (3459)
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.
BanII  (3385)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SgrAI  (2740)
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.
BbvCI  (173)
1 site
C C T C A G C G G A G T C G
Bpu10I  (173)
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.
BsmI  (823)
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.
XcmI  (931)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BlpI  (1209)
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.
EcoRI  (1217)
1 site
G A A T T C C T T A A G
BglII  (1223)
1 site
A G A T C T T C T A G A
PacI  (1270)
1 site
T T A A T T A A A A T T A A T T
EcoO109I  (1337)
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.
PpuMI  (1337)
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.
NsiI  (1358)
1 site
A T G C A T T A C G T A
XbaI  (1462)
1 site
T C T A G A A G A T C T
BclI  (1486)
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.
BsaBI  (1512)
1 site
G A T N N N N A T C C T A N N N N T A G
BamHI  (1582)
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.
SnaBI  (1652)
1 site
T A C G T A A T G C A T
SwaI  (1742)
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.
ZraI  (1931)
1 site
G A C G T C C T G C A G
AatII  (1933)
1 site
G A C G T C C T G C A G
HindIII  (2015)
1 site
A A G C T T T T C G A A
BmgBI  (2156)
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  (2500)
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  (2731)
1 site
C A C N N N N G T G G T G N N N N C A C
baculovirus recombination region (ORF1629)
1589 .. 2946  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
1589 .. 2946  =  1358 bp
contains part of ORF1629
AmpR
3803 .. 4663  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3803 .. 3871  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3803 .. 4663  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3872 .. 4663  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3803 .. 4663  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ORF603
100 .. 705  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
100 .. 705  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ori
4834 .. 5422  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4834 .. 5422  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 poly(A) signal
781 .. 902  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
781 .. 902  =  122 bp
SV40 polyadenylation signal
p10 promoter
1231 .. 1340  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
1231 .. 1340  =  110 bp
baculovirus promoter for expression in insect cells
AmpR promoter
3698 .. 3802  =  105 bp
AmpR promoter
3698 .. 3802  =  105 bp
polyhedrin promoter
1488 .. 1579  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
1488 .. 1579  =  92 bp
promoter for the baculovirus polyhedrin gene
ORF1629
1605 .. 2946  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
1605 .. 2946  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
baculovirus recombination region (ORF603)
178 .. 765  =  588 bp
contains ORF603 and part of lef2
baculovirus recombination region (ORF603)
178 .. 765  =  588 bp
contains ORF603 and part of lef2
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