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Plasmid Files

pDEST™17

Gateway® destination vector for inducible high-level expression of N-terminally 6xHis-tagged proteins in bacterial cells.

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pDEST17 Sequence and MappDEST17.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 XbaI (59) SgrAI (6314) SphI (6166) EcoNI (6101) PshAI (6008) NruI (5750) BstAPI (5676) BsgI * (5091) PfoI (4602) PflFI - Tth111I (4503) BsaAI (4497) BspQI - SapI (4362) AflIII - PciI (4245) AhdI (3357) NdeI (99) ATG 6xHis NotI (266) PasI (818) NcoI (887) BssHII (1080) BbvCI (1311) TspMI - XmaI (1455) SmaI - SrfI (1457) BmgBI (1491) BstXI (1574) SalI (1714) BlpI (1900) NheI (2128) BmtI (2132) HindIII (2328) BspDI - ClaI (2335) ZraI (2436) AatII (2438) pDEST™17 6354 bp
XbaI  (59)
1 site
T C T A G A A G A T C T
SgrAI  (6314)
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  (6166)
1 site
G C A T G C C G T A C G
EcoNI  (6101)
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.
PshAI  (6008)
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  (5750)
1 site
T C G C G A A G C G C T
BstAPI  (5676)
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.
BsgI  (5091)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
* Blocked by EcoKI methylation.
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).
PfoI  (4602)
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  (4503)
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  (4503)
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  (4497)
1 site
Y A C G T R R T G C A Y
BspQI  (4362)
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  (4362)
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.
AflIII  (4245)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4245)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (3357)
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.
NdeI  (99)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
NotI  (266)
1 site
G C G G C C G C C G C C G G C G
PasI  (818)
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.
NcoI  (887)
1 site
C C A T G G G G T A C C
BssHII  (1080)
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  (1311)
1 site
C C T C A G C G G A G T C G
TspMI  (1455)
1 site
C C C G G G G G G C C C
XmaI  (1455)
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  (1457)
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  (1457)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (1491)
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  (1574)
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  (1714)
1 site
G T C G A C C A G C T G
BlpI  (1900)
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  (2128)
1 site
G C T A G C C G A T C G
BmtI  (2132)
1 site
G C T A G C C G A T C G
HindIII  (2328)
1 site
A A G C T T T T C G A A
BspDI  (2335)
1 site
A T C G A T T A G C T A
ClaI  (2335)
1 site
A T C G A T T A G C T A
ZraI  (2436)
1 site
G A C G T C C T G C A G
AatII  (2438)
1 site
G A C G T C C T G C A G
AmpR
2570 .. 3430  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   2570 .. 2638  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2570 .. 3430  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2639 .. 3430  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2570 .. 3430  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
373 .. 1032  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
373 .. 1032  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3601 .. 4189  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3601 .. 4189  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ccdB
1374 .. 1679  =  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
1374 .. 1679  =  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
4619 .. 4810  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
4619 .. 4810  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
attR1
140 .. 264  =  125 bp
recombination site for the Gateway® LR reaction
attR1
140 .. 264  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1720 .. 1844  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1720 .. 1844  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
2465 .. 2569  =  105 bp
AmpR promoter
2465 .. 2569  =  105 bp
T7 terminator
1911 .. 1958  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
1911 .. 1958  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac UV5 promoter
289 .. 319  =  31 bp
   Segment 1:  -35  
   289 .. 294  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
289 .. 319  =  31 bp
   Segment 2:  
   295 .. 312  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
289 .. 319  =  31 bp
   Segment 3:  -10  
   313 .. 319  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
289 .. 319  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
113 .. 130  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
113 .. 130  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
86 .. 92  =  7 bp
ribosome binding site
RBS
86 .. 92  =  7 bp
ribosome binding site
ATG
101 .. 103  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
101 .. 103  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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