pEF-DEST51

Gateway® destination vector for high-level constitutive expression of C-terminally V5- and 6xHis-tagged proteins in mammalian cells.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspEF-DEST51
ZraI (119) FspI (6885) AhdI (6663) BspQI - SapI (5654) PfoI (5194) BlpI (5098) PflFI - Tth111I (4977) PpuMI (4862) AvrII (4574) CsiI - SexAI * (4341) PmeI (3525) SgrDI (120) AatII (121) AflII (854) FseI (1179) Eco53kI (1301) SacI (1303) KasI (1413) NarI (1414) SfoI (1415) PluTI (1417) PaeR7I - XhoI (1440) T7 promoter NotI (1846) BspEI (2162) EcoRI (2166) BsmBI - Esp3I (2390) PflMI * (2398) BssHII (2660) SrfI (3037) BmgBI (3071) BstXI (3154) XbaI * (3428) BstBI (3445) 6xHis pEF-DEST51 7464 bp
ZraI  (119)
1 site		 G A C G T C C T G C A G
FspI  (6885)
1 site		 T G C G C A A C G C G T
AhdI  (6663)
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.
BspQI  (5654)
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  (5654)
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.
PfoI  (5194)
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.
BlpI  (5098)
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.
PflFI  (4977)
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  (4977)
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.
PpuMI  (4862)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
AvrII  (4574)
1 site		 C C T A G G G G A T C C
CsiI  (4341)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (4341)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
PmeI  (3525)
1 site		 G T T T A A A C C A A A T T T G
SgrDI  (120)
1 site		 C G T C G A C G G C A G C T G C
AatII  (121)
1 site		 G A C G T C C T G C A G
AflII  (854)
1 site		 C T T A A G G A A T T C
FseI  (1179)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
Eco53kI  (1301)
1 site		 G A G C T C C T C G A G
SacI  (1303)
1 site		 G A G C T C C T C G A G
KasI  (1413)
1 site		 G G C G C C C C G C G G
NarI  (1414)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (1415)
1 site		 G G C G C C C C G C G G
PluTI  (1417)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
PaeR7I  (1440)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1440)
1 site		 C T C G A G G A G C T C
NotI  (1846)
1 site		 G C G G C C G C C G C C G G C G
BspEI  (2162)
1 site		 T C C G G A A G G C C T
EcoRI  (2166)
1 site		 G A A T T C C T T A A G
BsmBI  (2390)
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  (2390)
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.
PflMI  (2398)
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  (2660)
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.
SrfI  (3037)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (3071)
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  (3154)
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.
XbaI  (3428)
1 site		 T C T A G A A G A T C T
* Blocked by Dam methylation.
BstBI  (3445)
1 site		 T T C G A A A A G C T T
EF-1α promoter
475 .. 1653  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
475 .. 1653  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6590 .. 7381  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7382 .. 7450  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1953 .. 2612  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1953 .. 2612  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5831 .. 6419  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5831 .. 6419  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3817 .. 4245  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3817 .. 4245  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4703 .. 5101  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4703 .. 5101  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
4259 .. 4589  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
4259 .. 4589  =  331 bp
SV40 enhancer and early promoter
ccdB
2954 .. 3259  =  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
2954 .. 3259  =  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.
bGH poly(A) signal
3547 .. 3771  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
3547 .. 3771  =  225 bp
bovine growth hormone polyadenylation signal
attR1
1720 .. 1844  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1720 .. 1844  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3300 .. 3424  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3300 .. 3424  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5259 .. 5380  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5259 .. 5380  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7451 .. 91  =  105 bp
AmpR promoter
7451 .. 91  =  105 bp
V5 tag
3450 .. 3491  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3450 .. 3491  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
3501 .. 3518  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3501 .. 3518  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
EM7 promoter
4637 .. 4684  =  48 bp
synthetic bacterial promoter
EM7 promoter
4637 .. 4684  =  48 bp
synthetic bacterial promoter
lac UV5 promoter
1869 .. 1899  =  31 bp
3 segments
   Segment 1:  -35  
   1869 .. 1874  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1869 .. 1899  =  31 bp
3 segments
   Segment 2:  
   1875 .. 1892  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1869 .. 1899  =  31 bp
3 segments
   Segment 3:  -10  
   1893 .. 1899  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1869 .. 1899  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
1670 .. 1688  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1670 .. 1688  =  19 bp
promoter for bacteriophage T7 RNA polymerase
EF-1α intron A
706 .. 1644  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
706 .. 1644  =  939 bp
intron upstream of the start codon of human EF-1α
SV40 ori
4440 .. 4575  =  136 bp
SV40 origin of replication
SV40 ori
4440 .. 4575  =  136 bp
SV40 origin of replication
ORF:  1031 .. 1468  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  2954 .. 3259  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4703 .. 5101  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  1110 .. 1343  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  1953 .. 2612  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3768 .. 4034  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  6720 .. 6986  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  773 .. 1393  =  621 bp
ORF:  206 amino acids  =  23.0 kDa
ORF:  4649 .. 5005  =  357 bp
ORF:  118 amino acids  =  12.9 kDa
ORF:  6590 .. 7450  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pEF-DEST51.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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