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Plasmid Files

pDEST™20

Gateway® destination vector for expressing N-terminally GST-tagged proteins in insect cells using the Bac-to-Bac® baculovirus system.

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pDEST20 Sequence and MappDEST20.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 BbsI (6946) PflFI - Tth111I (6417) BseRI (6150) EcoRV (6014) SacII (5957) PspFI (5153) BseYI (5149) FspI (4342) PvuI (4196) BsaHI (4025) BanII (3350) SnaBI (4) EcoNI (171) EcoO109I (193) BsgI (447) BstBI (558) SwaI (588) NotI (968) PvuII (1188) EcoRI (1288) PasI - PflMI * (1520) NcoI (1589) XbaI (1735) BssHII (1782) BbvCI (2013) TspMI - XmaI (2157) SmaI - SrfI (2159) BmgBI (2193) BfuAI - BspMI (2403) PstI (2414) SalI (2416) attR2 MfeI (2666) HpaI (2679) AvrII (2830) NgoMIV (3316) NaeI (3318) pDEST™20 7066 bp
BbsI  (6946)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PflFI  (6417)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (6417)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BseRI  (6150)
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  (6014)
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.
SacII  (5957)
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.
PspFI  (5153)
1 site
C C C A G C G G G T C G
BseYI  (5149)
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.
FspI  (4342)
1 site
T G C G C A A C G C G T
PvuI  (4196)
1 site
C G A T C G G C T A G C
BsaHI  (4025)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
BanII  (3350)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SnaBI  (4)
1 site
T A C G T A A T G C A T
EcoNI  (171)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
EcoO109I  (193)
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.
BsgI  (447)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BstBI  (558)
1 site
T T C G A A A A G C T T
SwaI  (588)
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.
NotI  (968)
1 site
G C G G C C G C C G C C G G C G
PvuII  (1188)
1 site
C A G C T G G T C G A C
EcoRI  (1288)
1 site
G A A T T C C T T A A G
PasI  (1520)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (1520)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
NcoI  (1589)
1 site
C C A T G G G G T A C C
XbaI  (1735)
1 site
T C T A G A A G A T C T
BssHII  (1782)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BbvCI  (2013)
1 site
C C T C A G C G G A G T C G
TspMI  (2157)
1 site
C C C G G G G G G C C C
XmaI  (2157)
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  (2159)
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  (2159)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (2193)
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.
BfuAI  (2403)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (2403)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PstI  (2414)
1 site
C T G C A G G A C G T C
SalI  (2416)
1 site
G T C G A C C A G C T G
MfeI  (2666)
1 site
C A A T T G G T T A A C
HpaI  (2679)
1 site
G T T A A C C A A T T G
AvrII  (2830)
1 site
C C T A G G G G A T C C
NgoMIV  (3316)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (3318)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3778 .. 3846  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3847 .. 4638  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
1075 .. 1734  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1075 .. 1734  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
GST
161 .. 814  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from
Schistosoma japonicum
GST
161 .. 814  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from
Schistosoma japonicum
ori
4809 .. 5397  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4809 .. 5397  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
GmR
5991 .. 6524  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
5991 .. 6524  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
3191 .. 3646  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3191 .. 3646  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
ccdB
2076 .. 2381  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
2076 .. 2381  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
Tn7R
5700 .. 5924  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
5700 .. 5924  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
2843 .. 3008  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
2843 .. 3008  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
2680 .. 2814  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2680 .. 2814  =  135 bp
SV40 polyadenylation signal
attR1
842 .. 966  =  125 bp
recombination site for the Gateway® LR reaction
attR1
842 .. 966  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2422 .. 2546  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2422 .. 2546  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
3673 .. 3777  =  105 bp
AmpR promoter
3673 .. 3777  =  105 bp
polyhedrin promoter
27 .. 118  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
27 .. 118  =  92 bp
promoter for the baculovirus polyhedrin gene
lac promoter
991 .. 1021  =  31 bp
   Segment 1:  -35  
   991 .. 996  =  6 bp
promoter for the E. coli lac operon
lac promoter
991 .. 1021  =  31 bp
   Segment 2:  
   997 .. 1014  =  18 bp
promoter for the E. coli lac operon
lac promoter
991 .. 1021  =  31 bp
   Segment 3:  -10  
   1015 .. 1021  =  7 bp
promoter for the E. coli lac operon
lac promoter
991 .. 1021  =  31 bp
3 segments
promoter for the E. coli lac operon
Pc promoter
6713 .. 6741  =  29 bp
   Segment 3:  -10  
   6713 .. 6718  =  6 bp
class 1 integron promoter
Pc promoter
6713 .. 6741  =  29 bp
   Segment 2:  
   6719 .. 6735  =  17 bp
class 1 integron promoter
Pc promoter
6713 .. 6741  =  29 bp
   Segment 1:  -35  
   6736 .. 6741  =  6 bp
class 1 integron promoter
Pc promoter
6713 .. 6741  =  29 bp
3 segments
class 1 integron promoter
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