pEXP3-DEST

Gateway® destination vector for cell-free expression and N-terminal tagging. See also pEXP1-DEST.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspEXP3-DEST
BglII (0) BspQI - SapI (4545) NspI (4432) AflIII - PciI (4428) PspFI (4128) BseYI (4124) AhdI (3540) AfeI (3509) BmrI (3500) NmeAIII (3393) BsaHI (3000) XmnI (2940) PsiI (2470) DraIII (2345) BsaAI (2342) T7 promoter NdeI (98) ATG NsiI (104) 6xHis AleI (119) XcmI (124) TEV site EagI - NotI (313) lac UV5 promoter PvuII (533) EcoRI (633) BsmBI - Esp3I (857) PasI - PflMI * (865) BssHII (1127) BstZ17I (1168) BbvCI (1358) AvaI - BsoBI - TspMI - XmaI (1502) SmaI - SrfI (1504) BmgBI (1538) BfuAI - BspMI (1748) PstI (1759) SalI (1761) HindIII (1899) BlpI (1957) EcoO109I (1984) T7 terminator NgoMIV (2237) NaeI (2239) BanII (2271) pEXP3-DEST 4607 bp
BglII  (0)
1 site		 A G A T C T T C T A G A
BspQI  (4545)
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  (4545)
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  (4432)
1 site		 R C A T G Y Y G T A C R
AflIII  (4428)
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  (4428)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PspFI  (4128)
1 site		 C C C A G C G G G T C G
BseYI  (4124)
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.
AhdI  (3540)
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.
AfeI  (3509)
1 site		 A G C G C T T C G C G A
BmrI  (3500)
1 site		 A C T G G G ( N ) 4 N T G A C C C ( N ) 4
The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
NmeAIII  (3393)
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).
BsaHI  (3000)
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.
XmnI  (2940)
1 site		 G A A N N N N T T C C T T N N N N A A G
PsiI  (2470)
1 site		 T T A T A A A A T A T T
DraIII  (2345)
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.
BsaAI  (2342)
1 site		 Y A C G T R R T G C A Y
NdeI  (98)
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.
NsiI  (104)
1 site		 A T G C A T T A C G T A
AleI  (119)
1 site		 C A C N N N N G T G G T G N N N N C A C
XcmI  (124)
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.
EagI  (313)
1 site		 C G G C C G G C C G G C
NotI  (313)
1 site		 G C G G C C G C C G C C G G C G
PvuII  (533)
1 site		 C A G C T G G T C G A C
EcoRI  (633)
1 site		 G A A T T C C T T A A G
BsmBI  (857)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (857)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
PasI  (865)
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  (865)
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.
BssHII  (1127)
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.
BstZ17I  (1168)
1 site		 G T A T A C C A T A T G
BbvCI  (1358)
1 site		 C C T C A G C G G A G T C G
AvaI  (1502)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1502)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (1502)
1 site		 C C C G G G G G G C C C
XmaI  (1502)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (1504)
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  (1504)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (1538)
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  (1748)
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  (1748)
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  (1759)
1 site		 C T G C A G G A C G T C
SalI  (1761)
1 site		 G T C G A C C A G C T G
HindIII  (1899)
1 site		 A A G C T T T T C G A A
BlpI  (1957)
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.
EcoO109I  (1984)
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.
NgoMIV  (2237)
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  (2239)
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.
BanII  (2271)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
AmpR
2753 .. 3613  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2753 .. 2821  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2753 .. 3613  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2822 .. 3613  =  792 bp
   263 amino acids  =  29.0 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2753 .. 3613  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
420 .. 1079  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
420 .. 1079  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3784 .. 4372  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3784 .. 4372  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2112 .. 2567  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2112 .. 2567  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
ccdB
1421 .. 1726  =  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
1421 .. 1726  =  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.
attR1
187 .. 311  =  125 bp
recombination site for the Gateway® LR reaction
attR1
187 .. 311  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1767 .. 1891  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1767 .. 1891  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
2648 .. 2752  =  105 bp
AmpR promoter
2648 .. 2752  =  105 bp
ATG
100 .. 102  =  3 bp
1 amino acid  =  149.2 Da
ATG
100 .. 102  =  3 bp
1 amino acid  =  149.2 Da
6xHis
103 .. 120  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
103 .. 120  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
tetracysteine tag
133 .. 150  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
tetracysteine tag
133 .. 150  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
TEV site
160 .. 180  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
160 .. 180  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
T7 terminator
1968 .. 2015  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1968 .. 2015  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac UV5 promoter
336 .. 366  =  31 bp
3 segments
   Segment 1:  -35  
   336 .. 341  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
336 .. 366  =  31 bp
3 segments
   Segment 2:  
   342 .. 359  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
336 .. 366  =  31 bp
3 segments
   Segment 3:  -10  
   360 .. 366  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
336 .. 366  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
RBS
70 .. 92  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
70 .. 92  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
20 .. 38  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
20 .. 38  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1421 .. 1726  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2753 .. 3613  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  420 .. 1079  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3217 .. 3483  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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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

Plasmid Sets

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I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
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Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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