pAcUW51

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

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

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.
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
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
2 segments
   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
2 segments
   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
ORF:  3803 .. 4663  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  689 .. 1114  =  426 bp
ORF:  141 amino acids  =  16.6 kDa
ORF:  100 .. 705  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  4267 .. 4533  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  144 .. 374  =  231 bp
ORF:  76 amino acids  =  9.3 kDa
ORF:  1605 .. 2858  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
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