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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HomePlasmidsMammalian Expression 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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Individual Sequences & Maps

p3xFLAG-CMV-10
pcDNA6 myc-His A
pCMV6-AC-YFP
pHet-Act1-2
p3xFLAG-CMV-13
pcDNA6 myc-His B
pCMV6-AN-3DDK
pHet-Mem1
p3xFLAG-CMV-14
pcDNA6 myc-His C
pCMV6-AN-3HA
pHet-Nuc1
p3xFLAG-CMV-7
pcDNA6 TR
pCMV6-AN-DDK
pHom-1
p3xFLAG-CMV-7.1
pcDNA6 V5-His A
pCMV6-AN-FC
pHom-Mem1
p3xFLAG-CMV-8
pcDNA6 V5-His B
pCMV6-AN-FC-S
pHTC HaloTag CMV-neo
p3xFLAG-CMV-9
pcDNA6 V5-His C
pCMV6-AN-FP602
pHTN HaloTag CMV-neo
p3xFLAG-Myc-CMV-23
pCEP4
pCMV6-AN-FP635
pIRES
p3xFLAG-Myc-CMV-24
pCI
pCMV6-AN-GFP
pIRES2 DsRed-Express2
p3xFLAG-Myc-CMV-25
pCI-neo
pCMV6-AN-GFP-AC-His
pIRES2-AcGFP1
p3xFLAG-Myc-CMV-26
pCMV SPORT6
pCMV6-AN-HA
pIRES2-EGFP
pALTER-MAX
pCMV SPORT6.1
pCMV6-AN-His
pIRES2-ZsGreen1
pBApo-CMV
pCMV-(DYKDDDDK)-C
pCMV6-AN-His-DDK
pIRESbleo3
pBApo-CMV Neo
pCMV-(DYKDDDDK)-N
pCMV6-AN-His-HA
pIREShyg3
pBApo-CMV Pur
pCMV-3Tag-1A
pCMV6-AN-His-Myc
pIRESneo3
pBApo-EF1(alpha) Neo
pCMV-3Tag-1B
pCMV6-AN-mBFP
pIRESpuro3
pBApo-EF1(alpha) Pur
pCMV-3Tag-1C
pCMV6-AN-mCFP
pM-secSUMOstar
pBI-CMV1
pCMV-3Tag-2A
pCMV6-AN-mGFP
pM-SUMOstar
pBI-CMV2
pCMV-3Tag-2B
pCMV6-AN-mKate
pME-HA (linearized)
pBI-CMV3
pCMV-3Tag-2C
pCMV6-AN-mRFP
pmRi-mCherry
pBI-CMV4
pCMV-3Tag-3A
pCMV6-AN-Myc
pmRi-ZsGreen1
pBK-CMV
pCMV-3Tag-3B
pCMV6-AN-Myc-DDK
pNTAP-A
pBudCE4.1
pCMV-3Tag-3C
pCMV6-AN-mYFP
pNTAP-B
pcDNA3
pCMV-3Tag-4A
pCMV6-AN-RFP
pNTAP-C
pcDNA3.1+ C-DYK
pCMV-3Tag-4B
pCMV6-AN-YFP
pOG44
pcDNA3.1+ C-HA
pCMV-3Tag-4C
pCMV6-Entry
pPTuner
pcDNA3.1+ C-His
pCMV-3Tag-6
pCMV6-Entry-Cre
pPTuner IRES2
pcDNA3.1+ C-Myc
pCMV-3Tag-7
pCMV6-Entry2
pPTunerC
pcDNA3.1+ N-GST(TEV)
pCMV-3Tag-8
pCMV6-XL5-DDK-His
pREP4
pcDNA3.1+ N-GST(Thrombin)
pCMV-3Tag-9
pCS6-Empty
prHom-1
pcDNA3.1+ N-HA
pCMV-DDK-C
pCTAP-A
prHom-Nuc1
pcDNA3.1+ N-His
pCMV-DDK-N
pCTAP-B
prHom-Sec1
pcDNA3.1+ N-Myc
pCMV-HA
pCTAP-C
pSecTag A
pcDNA-DEST40
pCMV-HA-C
pDEST26
pSecTag B
pcDNA-DEST47
pCMV-HA-N
pDEST27
pSecTag C
pcDNA-DEST53
pCMV-LacZ
pDisplay
pSecTag2 A
pcDNA3.1(+)
pCMV-MIR
pEF-DEST51
pSecTag2 B
pcDNA3.1(-)
pCMV-Myc
pEF1alpha-IRES
pSecTag2 C
pcDNA3.1 CT-GFP
pCMV-Myc-C
pEF1alpha-IRES-AcGFP1
pSI
pcDNA3.1 His A
pCMV-Myc-N
pEF1alpha-IRES-DsRed-Express2
pSingle-tTs-shRNA
pcDNA3.1 His B
pCMV-Script
pEF1alpha-IRES-ZsGreen1
pT-REx-DEST30
pcDNA3.1 His C
pCMV-Script EX
pEF1alpha-Tet3G
pT-REx-DEST31
pcDNA3.1 Hygro(+)
pCMV-Tag 1
pEF5 FRT V5-DEST
pTandem-1
pcDNA3.1 Hygro(-)
pCMV-Tag 2A
pEF6 V5-His A
pTCN
pcDNA3.1 myc-His A
pCMV-Tag 2B
pEF6 V5-His B
pTCP
pcDNA3.1 myc-His B
pCMV-Tag 2C
pEF6 V5-His C
pTet-DualOFF
pcDNA3.1 myc-His C
pCMV-Tag 3A
pEntry-rtTA
pTet-DualON
pcDNA3.1 myc-His(-) A
pCMV-Tag 3B
pF4A CMV
pTet-Off
pcDNA3.1 myc-His(-) B
pCMV-Tag 3C
pF4K CMV
pTet-Off-Advanced
pcDNA3.1 myc-His(-) C
pCMV-Tag 4A
pF5A CMV-neo
pTet-On
pcDNA3.1 NT-GFP
pCMV-Tag 4B
pF5K CMV-neo
pTet-On-Advanced
pcDNA3.1 nV5-DEST
pCMV-Tag 4C
pFC14A HaloTag CMV
pTet-tTS
pcDNA3.1 V5-His A
pCMV-Tag 5A
pFC14K HaloTag CMV
pTetOne
pcDNA3.1 V5-His B
pCMV-Tag 5B
pFC15A HaloTag CMVd1
pTK-neo
pcDNA3.1 V5-His C
pCMV-Tag 5C
pFC15K HaloTag CMVd1
pTracer-CMV
pcDNA3.1 Zeo(+)
pCMV-Tet3G
pFC16A HaloTag CMVd2
pTracer-CMV Bsd
pcDNA3.1 Zeo(-)
pCMV6-A-BSD
pFC16K HaloTag CMVd2
pTracer-CMV2
pcDNA3.2 V5-DEST
pCMV6-A-EM7-BSD
pFC17A HaloTag CMVd3
pTracer-EF V5-His A
pcDNA4 His A
pCMV6-A-GFP
pFC17K HaloTag CMVd3
pTracer-EF V5-His B
pcDNA4 His B
pCMV6-A-Hygro
pFC27A HaloTag CMV-neo
pTracer-EF V5-His C
pcDNA4 His C
pCMV6-A-Puro
pFC27K HaloTag CMV-neo
pTracer-SV40
pcDNA4 HisMax A
pCMV6-AC
pFLAG-CMV-1
pTRE-Cycle1
pcDNA4 HisMax B
pCMV6-AC-3DDK
pFLAG-CMV-2
pTRE-Cycle2
pcDNA4 HisMax C
pCMV6-AC-3HA
pFLAG-CMV-3
pTRE-Cycle3
pcDNA4 myc-His A
pCMV6-AC-DDK
pFLAG-CMV-4
pTRE-Dual1
pcDNA4 myc-His B
pCMV6-AC-DDK-His
pFLAG-CMV-5.1
pTRE-Dual2
pcDNA4 myc-His C
pCMV6-AC-FC
pFLAG-Myc-CMV-19
pTRE-Tight
pcDNA4 TO
pCMV6-AC-FC-S
pFLAG-Myc-CMV-20
pTRE-Tight-BI
pcDNA4 TO myc-His A
pCMV6-AC-FP602
pFLAG-Myc-CMV-21
pTRE-Tight-BI-AcGFP1
pcDNA4 TO myc-His B
pCMV6-AC-FP635
pFLAG-Myc-CMV-22
pTRE-Tight-BI-DsRed-Express
pcDNA4 TO myc-His C
pCMV6-AC-GFP
pFN21A HaloTag CMV
pTRE-Tight-BI-ZsGreen1
pcDNA4 V5-His A
pCMV6-AC-GFP-rtTA
pFN21K HaloTag CMV
pTRE2
pcDNA4 V5-His B
pCMV6-AC-HA
pFN22A HaloTag CMVd1
pTRE2hyg
pcDNA4 V5-His C
pCMV6-AC-HA-His
pFN22K HaloTag CMVd1
pTRE2pur
pcDNA5 FRT
pCMV6-AC-His
pFN23A HaloTag CMVd2
pTRE3G
pcDNA5 FRT TO
pCMV6-AC-IRES-GFP
pFN23K HaloTag CMVd2
pTRE3G-BI
pcDNA5 TO
pCMV6-AC-IRES-GFP-Puro
pFN24A HaloTag CMVd3
pTRE3G-BI-Het1
pcDNA6.2 C-EmGFP-DEST
pCMV6-AC-mBFP
pFN24K HaloTag CMVd3
pTRE3G-BI-mCherry
pcDNA6.2 C-YFP-DEST
pCMV6-AC-mCFP
pFN28A HaloTag CMV-neo
pTRE3G-BI-ZsGreen1
pcDNA6.2 cGeneBLAzer-DEST
pCMV6-AC-mGFP
pFN28K HaloTag CMV-neo
pTRE3G-IRES
pcDNA6.2 cLumio-DEST
pCMV6-AC-mKate
pFRT lacZeo
pTRE3G-mCherry
pcDNA6.2 EmGFP-Bsd V5-DEST
pCMV6-AC-mRFP
pFRT lacZeo2
pTRE3G-ZsGreen1
pcDNA6.2 N-EmGFP-DEST
pCMV6-AC-Myc
pGen2.1
pVAX1
pcDNA6.2 N-YFP-DEST
pCMV6-AC-Myc-DDK
phCMV1
pX330
pcDNA6.2 nGeneBLAzer-DEST
pCMV6-AC-Myc-His
phCMV2
pZeoSV2(+)
pcDNA6.2 nLumio-DEST
pCMV6-AC-mYFP
phCMV3
pZeoSV2(-)
pcDNA6.2 V5-DEST
pCMV6-AC-RFP
pHet-1
pZFHD1-2
pcDNA6.2 V5-pL-DEST

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
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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