pFLAG-CMV-3

Vector with a neomycin resistance marker for expression and secretion of N-terminally FLAG-tagged proteins in mammalian cells.

Sequence Author: MilliporeSigma

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HomePlasmidsMammalian Expression VectorspFLAG-CMV-3
ScaI (5420) AflIII - PciI (4047) HpaI (3533) MfeI (3520) AfeI (3328) BsrGI (276) SpeI (332) CMV enhancer NdeI (567) SnaBI (673) Eco53kI (899) SacI (901) HindIII (1008) NotI (1015) EcoRI (1022) BglII (1034) EcoRV (1042) Acc65I (1046) KpnI (1050) XbaI (1059) BamHI (1065) AleI (1129) EcoO109I (1259) BbvCI - Bpu10I (1355) BlpI (1399) BsmBI - Esp3I (1419) BstXI (1506) AgeI (1617) PflMI (1620) XcmI (1712) CsiI - SexAI * (1804) SfiI (1991) BseRI (2034) PflFI - Tth111I (2334) BssHII (2613) RsrII (2732) BstBI (2898) NruI (3071) pFLAG-CMV™-3 6331 bp
ScaI  (5420)
1 site		 A G T A C T T C A T G A
AflIII  (4047)
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  (4047)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
HpaI  (3533)
1 site		 G T T A A C C A A T T G
MfeI  (3520)
1 site		 C A A T T G G T T A A C
AfeI  (3328)
1 site		 A G C G C T T C G C G A
BsrGI  (276)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (332)
1 site		 A C T A G T T G A T C A
NdeI  (567)
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  (673)
1 site		 T A C G T A A T G C A T
Eco53kI  (899)
1 site		 G A G C T C C T C G A G
SacI  (901)
1 site		 G A G C T C C T C G A G
HindIII  (1008)
1 site		 A A G C T T T T C G A A
NotI  (1015)
1 site		 G C G G C C G C C G C C G G C G
EcoRI  (1022)
1 site		 G A A T T C C T T A A G
BglII  (1034)
1 site		 A G A T C T T C T A G A
EcoRV  (1042)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
Acc65I  (1046)
1 site		 G G T A C C C C A T G G
KpnI  (1050)
1 site		 G G T A C C C C A T G G
XbaI  (1059)
1 site		 T C T A G A A G A T C T
BamHI  (1065)
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.
AleI  (1129)
1 site		 C A C N N N N G T G G T G N N N N C A C
EcoO109I  (1259)
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.
BbvCI  (1355)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (1355)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BlpI  (1399)
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.
BsmBI  (1419)
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  (1419)
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.
BstXI  (1506)
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.
AgeI  (1617)
1 site		 A C C G G T T G G C C A
PflMI  (1620)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
XcmI  (1712)
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.
CsiI  (1804)
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  (1804)
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.
SfiI  (1991)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BseRI  (2034)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
PflFI  (2334)
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  (2334)
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.
BssHII  (2613)
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.
RsrII  (2732)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstBI  (2898)
1 site		 T T C G A A A A G C T T
NruI  (3071)
1 site		 T C G C G A A G C G C T
AmpR
4867 .. 5727  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4867 .. 5658  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4867 .. 5727  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5659 .. 5727  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4867 .. 5727  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2088 .. 2882  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2088 .. 2882  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
hGH poly(A) signal
1072 .. 1694  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1072 .. 1694  =  623 bp
human growth hormone polyadenylation signal
ori
4108 .. 4696  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4108 .. 4696  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5859 .. 6331  =  473 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5859 .. 6331  =  473 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
318 .. 697  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
318 .. 697  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
1736 .. 2037  =  302 bp
SV40 enhancer and early promoter
SV40 promoter
1736 .. 2037  =  302 bp
SV40 enhancer and early promoter
CMV promoter
698 .. 901  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
698 .. 901  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
3534 .. 3668  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3534 .. 3668  =  135 bp
SV40 polyadenylation signal
AmpR promoter
5728 .. 5832  =  105 bp
AmpR promoter
5728 .. 5832  =  105 bp
preprotrypsin leader
942 .. 986  =  45 bp
15 amino acids  =  1.4 kDa
Product: leader sequence from mouse preprotrypsin
preprotrypsin leader
942 .. 986  =  45 bp
15 amino acids  =  1.4 kDa
Product: leader sequence from mouse preprotrypsin
FLAG
987 .. 1010  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG epitope tag, followed by an enterokinase cleavage site
FLAG
987 .. 1010  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG epitope tag, followed by an enterokinase cleavage site
MCS
1008 .. 1074  =  67 bp
multiple cloning site
MCS
1008 .. 1074  =  67 bp
multiple cloning site
ORF:  2260 .. 2646  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2903 .. 3283  =  381 bp
ORF:  126 amino acids  =  14.1 kDa
ORF:  4997 .. 5263  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2088 .. 2882  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1436 .. 1660  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  2933 .. 3178  =  246 bp
ORF:  81 amino acids  =  8.4 kDa
ORF:  3163 .. 3417  =  255 bp
ORF:  84 amino acids  =  9.3 kDa
ORF:  4867 .. 5727  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2397 .. 2933  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3081 .. 3443  =  363 bp
ORF:  120 amino acids  =  13.1 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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