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Plasmid Files

pDEST™24

Gateway® destination vector for inducible high-level expression of C-terminally GST-tagged proteins in bacterial cells.

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pDEST24.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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SgrAI (6925) SphI (6777) PshAI (6619) NruI (6361) BstAPI (6287) PfoI (5213) PflFI - Tth111I (5114) BsaAI (5108) AhdI (3968) BglII (5) XbaI * (63) NotI (197) PasI (749) MluI (964) BssHII (1011) BbvCI (1242) TspMI - XmaI (1386) SmaI - SrfI (1388) BmgBI (1422) BstXI (1505) SalI (1645) BstBI (2177) SwaI (2207) BclI * (2214) BlpI (2511) NheI (2739) BmtI (2743) HindIII (2939) BspDI - ClaI (2946) ZraI (3047) AatII (3049) pDEST™24 6961 bp
SgrAI  (6925)
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.
SphI  (6777)
1 site
G C A T G C C G T A C G
PshAI  (6619)
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.
NruI  (6361)
1 site
T C G C G A A G C G C T
BstAPI  (6287)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PfoI  (5213)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
PflFI  (5114)
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  (5114)
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.
BsaAI  (5108)
1 site
Y A C G T R R T G C A Y
AhdI  (3968)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BglII  (5)
1 site
A G A T C T T C T A G A
XbaI  (63)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
NotI  (197)
1 site
G C G G C C G C C G C C G G C G
PasI  (749)
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.
MluI  (964)
1 site
A C G C G T T G C G C A
BssHII  (1011)
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  (1242)
1 site
C C T C A G C G G A G T C G
TspMI  (1386)
1 site
C C C G G G G G G C C C
XmaI  (1386)
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  (1388)
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  (1388)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (1422)
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.
BstXI  (1505)
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.
SalI  (1645)
1 site
G T C G A C C A G C T G
BstBI  (2177)
1 site
T T C G A A A A G C T T
SwaI  (2207)
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.
BclI  (2214)
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.
BlpI  (2511)
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.
NheI  (2739)
1 site
G C T A G C C G A T C G
BmtI  (2743)
1 site
G C T A G C C G A T C G
HindIII  (2939)
1 site
A A G C T T T T C G A A
BspDI  (2946)
1 site
A T C G A T T A G C T A
ClaI  (2946)
1 site
A T C G A T T A G C T A
ZraI  (3047)
1 site
G A C G T C C T G C A G
AatII  (3049)
1 site
G A C G T C C T G C A G
AmpR
3181 .. 4041  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3181 .. 3249  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3181 .. 4041  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3250 .. 4041  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3181 .. 4041  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
304 .. 963  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
304 .. 963  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
GST
1780 .. 2433  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
1780 .. 2433  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
ori
4212 .. 4800  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4212 .. 4800  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
1305 .. 1610  =  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
1305 .. 1610  =  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.
rop
5230 .. 5421  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
5230 .. 5421  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
attR1
71 .. 195  =  125 bp
recombination site for the Gateway® LR reaction
attR1
71 .. 195  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1651 .. 1775  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1651 .. 1775  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
3076 .. 3180  =  105 bp
AmpR promoter
3076 .. 3180  =  105 bp
T7 terminator
2522 .. 2569  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2522 .. 2569  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac UV5 promoter
220 .. 250  =  31 bp
   Segment 1:  -35  
   220 .. 225  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
220 .. 250  =  31 bp
   Segment 2:  
   226 .. 243  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
220 .. 250  =  31 bp
   Segment 3:  -10  
   244 .. 250  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
220 .. 250  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
25 .. 43  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
25 .. 43  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  304 .. 963  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  1780 .. 2472  =  693 bp
ORF:  230 amino acids  =  26.8 kDa
ORF:  3181 .. 4041  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6775 .. 116  =  303 bp
ORF:  100 amino acids  =  10.6 kDa
ORF:  5453 .. 5821  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  1305 .. 1610  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  6255 .. 6740  =  486 bp
ORF:  161 amino acids  =  17.4 kDa
ORF:  2522 .. 2887  =  366 bp
ORF:  121 amino acids  =  12.9 kDa
ORF:  5230 .. 5454  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  5818 .. 6156  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  6262 .. 6849  =  588 bp
ORF:  195 amino acids  =  21.5 kDa
ORF:  3645 .. 3911  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6060 .. 6677  =  618 bp
ORF:  205 amino acids  =  21.8 kDa
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