pcDNA6.2 V5-DEST

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsMammalian Expression VectorspcDNA6.2 V5-DEST
SgrDI (7339) PciI (5528) BspQI - SapI (5412) PfoI (4952) BlpI (4856) BsgI (4648) PpuMI (4620) EagI (4577) BbsI (4546) BclI * (4383) AvrII (4332) StuI (4331) CsiI - SexAI * (4099) MfeI (161) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) PstI (1434) BfuAI - BspMI (1437) BstXI (1572) BmgBI (1651) SrfI (1685) BbvCI (1828) BssHII (2058) BamHI (2099) EcoRI (2552) BspEI (2556) XbaI * (3179) PspOMI (3185) ApaI (3189) SacII (3192) BstBI (3196) AgeI (3246) PmeI (3264) pcDNA™6.2/V5-DEST 7341 bp
SgrDI  (7339)
1 site		 C G T C G A C G G C A G C T G C
PciI  (5528)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (5412)
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  (5412)
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  (4952)
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  (4856)
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.
BsgI  (4648)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
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).
PpuMI  (4620)
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.
EagI  (4577)
1 site		 C G G C C G G C C G G C
BbsI  (4546)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BclI  (4383)
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.
AvrII  (4332)
1 site		 C C T A G G G G A T C C
StuI  (4331)
1 site		 A G G C C T T C C G G A
CsiI  (4099)
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  (4099)
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.
MfeI  (161)
1 site		 C A A T T G G T T A A C
SpeI  (249)
1 site		 A C T A G T T G A T C A
NdeI  (484)
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.
SnaBI  (590)
1 site		 T A C G T A A T G C A T
Eco53kI  (816)
1 site		 G A G C T C C T C G A G
SacI  (818)
1 site		 G A G C T C C T C G A G
PstI  (1434)
1 site		 C T G C A G G A C G T C
BfuAI  (1437)
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  (1437)
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.
BstXI  (1572)
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.
BmgBI  (1651)
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.
SrfI  (1685)
1 site		 G C C C G G G C C G G G C C C G
BbvCI  (1828)
1 site		 C C T C A G C G G A G T C G
BssHII  (2058)
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.
BamHI  (2099)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRI  (2552)
1 site		 G A A T T C C T T A A G
BspEI  (2556)
1 site		 T C C G G A A G G C C T
XbaI  (3179)
1 site		 T C T A G A A G A T C T
* Blocked by Dam methylation.
PspOMI  (3185)
1 site		 G G G C C C C C C G G G
ApaI  (3189)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
SacII  (3192)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
BstBI  (3196)
1 site		 T T C G A A A A G C T T
AgeI  (3246)
1 site		 A C C G G T T G G C C A
PmeI  (3264)
1 site		 G T T T A A A C C A A A T T T G
AmpR
6345 .. 7205  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6345 .. 7136  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6345 .. 7205  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7137 .. 7205  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6345 .. 7205  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
2111 .. 2770  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2111 .. 2770  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5589 .. 6174  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5589 .. 6174  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3576 .. 4004  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3576 .. 4004  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4461 .. 4859  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4461 .. 4859  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
4018 .. 4347  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4018 .. 4347  =  330 bp
SV40 enhancer and early promoter
ccdB
1464 .. 1769  =  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
1464 .. 1769  =  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.
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3051 .. 3175  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3051 .. 3175  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5017 .. 5138  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5017 .. 5138  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7206 .. 7310  =  105 bp
AmpR promoter
7206 .. 7310  =  105 bp
cat promoter
2771 .. 2873  =  103 bp
promoter of the E. coli cat gene
cat promoter
2771 .. 2873  =  103 bp
promoter of the E. coli cat gene
HSV TK poly(A) signal
3326 .. 3374  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3326 .. 3374  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
4395 .. 4442  =  48 bp
synthetic bacterial promoter
EM7 promoter
4395 .. 4442  =  48 bp
synthetic bacterial promoter
V5 tag
3201 .. 3242  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3201 .. 3242  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
4198 .. 4333  =  136 bp
SV40 origin of replication
SV40 ori
4198 .. 4333  =  136 bp
SV40 origin of replication
ORF:  6475 .. 6741  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4461 .. 4859  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  1464 .. 1769  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4407 .. 4763  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  6345 .. 7205  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2111 .. 2770  =  660 bp
ORF:  219 amino acids  =  25.7 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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