pDEST15

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspDEST15
SgrAI (6976) SphI (6828) PshAI (6670) NruI (6412) BstAPI (6338) PfoI (5264) PflFI - Tth111I (5165) BsaAI (5159) AhdI (4019) BglII (5) NdeI (103) BstBI (502) SwaI (532) BclI * (539) NotI (918) PasI (1470) BssHII (1731) BbvCI (1962) SrfI (2108) BmgBI (2142) BstXI (2225) SalI (2365) BlpI (2562) NheI (2790) BmtI (2794) HindIII (2990) BspDI - ClaI (2997) ZraI (3098) AatII (3100) pDEST™15 7012 bp
SgrAI  (6976)
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  (6828)
1 site		 G C A T G C C G T A C G
PshAI  (6670)
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  (6412)
1 site		 T C G C G A A G C G C T
BstAPI  (6338)
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  (5264)
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  (5165)
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  (5165)
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  (5159)
1 site		 Y A C G T R R T G C A Y
AhdI  (4019)
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
NdeI  (103)
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.
BstBI  (502)
1 site		 T T C G A A A A G C T T
SwaI  (532)
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  (539)
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.
NotI  (918)
1 site		 G C G G C C G C C G C C G G C G
PasI  (1470)
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.
BssHII  (1731)
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  (1962)
1 site		 C C T C A G C G G A G T C G
SrfI  (2108)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (2142)
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  (2225)
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  (2365)
1 site		 G T C G A C C A G C T G
BlpI  (2562)
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  (2790)
1 site		 G C T A G C C G A T C G
BmtI  (2794)
1 site		 G C T A G C C G A T C G
HindIII  (2990)
1 site		 A A G C T T T T C G A A
BspDI  (2997)
1 site		 A T C G A T T A G C T A
ClaI  (2997)
1 site		 A T C G A T T A G C T A
ZraI  (3098)
1 site		 G A C G T C C T G C A G
AatII  (3100)
1 site		 G A C G T C C T G C A G
AmpR
3232 .. 4092  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   3232 .. 3300  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3232 .. 4092  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   3301 .. 4092  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3232 .. 4092  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1025 .. 1705  =  681 bp
226 amino acids  =  26.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1025 .. 1705  =  681 bp
226 amino acids  =  26.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
GST
105 .. 758  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
105 .. 758  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
ori
4263 .. 4851  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4263 .. 4851  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
2025 .. 2330  =  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
2025 .. 2330  =  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
5281 .. 5472  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
5281 .. 5472  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
attR1
792 .. 916  =  125 bp
recombination site for the Gateway® LR reaction
attR1
792 .. 916  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2371 .. 2495  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2371 .. 2495  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
3127 .. 3231  =  105 bp
AmpR promoter
3127 .. 3231  =  105 bp
T7 terminator
2573 .. 2620  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2573 .. 2620  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac UV5 promoter
941 .. 971  =  31 bp
3 segments
   Segment 1:  -35  
   941 .. 946  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
941 .. 971  =  31 bp
3 segments
   Segment 2:  
   947 .. 964  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
941 .. 971  =  31 bp
3 segments
   Segment 3:  -10  
   965 .. 971  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
941 .. 971  =  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
RBS
90 .. 96  =  7 bp
ribosome binding site
RBS
90 .. 96  =  7 bp
ribosome binding site
ORF:  3232 .. 4092  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6826 .. 83  =  270 bp
ORF:  89 amino acids  =  9.3 kDa
ORF:  1025 .. 1705  =  681 bp
ORF:  226 amino acids  =  26.6 kDa
ORF:  5504 .. 5872  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  105 .. 827  =  723 bp
ORF:  240 amino acids  =  28.1 kDa
ORF:  2025 .. 2330  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  6306 .. 6791  =  486 bp
ORF:  161 amino acids  =  17.4 kDa
ORF:  2573 .. 2938  =  366 bp
ORF:  121 amino acids  =  12.9 kDa
ORF:  5281 .. 5505  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  5869 .. 6207  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  6313 .. 6900  =  588 bp
ORF:  195 amino acids  =  21.5 kDa
ORF:  3696 .. 3962  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6111 .. 6728  =  618 bp
ORF:  205 amino acids  =  21.8 kDa
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Download pDEST15.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

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