pFN21A HaloTag CMV

Flexi® vector with an ampicillin resistance marker, for mammalian expression of a protein with a cleavable N-terminal HaloTag®.

Sequence Author: Promega

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HomePlasmidsMammalian Expression VectorspFN21A HaloTag CMV
BsaBI * (5058) MluI (4940) BspEI (4727) PciI (3963) AgeI (3895) BstBI (3868) AhdI (3793) FspI (3570) XmnI (3193) MfeI (2663) HpaI (2654) AanI - PsiI (2634) BglII (5059) BsrGI (96) SpeI (152) NdeI (387) HindIII (748) BstAPI (794) T7 promoter NheI (1052) BmtI (1056) PflMI (1108) AleI (1166) BstEII (1184) BseRI (1186) PflFI - Tth111I (1316) BmgBI (1318) PasI (1348) StuI * (1517) BclI * (1548) BspDI * - ClaI * (1554) PshAI (1613) SgrAI (1700) BssHII (1925) PaeR7I - XhoI (1943) AsiSI - PvuI - SgfI (2001) DraI - PmeI (2368) Acc65I (2385) KpnI (2389) XbaI (2400) SbfI (2416) BlpI (2481) NotI (2511) pFN21A HaloTag® CMV 5064 bp
BsaBI  (5058)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MluI  (4940)
1 site		 A C G C G T T G C G C A
BspEI  (4727)
1 site		 T C C G G A A G G C C T
PciI  (3963)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AgeI  (3895)
1 site		 A C C G G T T G G C C A
BstBI  (3868)
1 site		 T T C G A A A A G C T T
AhdI  (3793)
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.
FspI  (3570)
1 site		 T G C G C A A C G C G T
XmnI  (3193)
1 site		 G A A N N N N T T C C T T N N N N A A G
MfeI  (2663)
1 site		 C A A T T G G T T A A C
HpaI  (2654)
1 site		 G T T A A C C A A T T G
AanI  (2634)
1 site		 T T A T A A A A T A T T
PsiI  (2634)
1 site		 T T A T A A A A T A T T
BglII  (5059)
1 site		 A G A T C T T C T A G A
BsrGI  (96)
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  (152)
1 site		 A C T A G T T G A T C A
NdeI  (387)
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.
HindIII  (748)
1 site		 A A G C T T T T C G A A
BstAPI  (794)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
NheI  (1052)
1 site		 G C T A G C C G A T C G
BmtI  (1056)
1 site		 G C T A G C C G A T C G
PflMI  (1108)
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.
AleI  (1166)
1 site		 C A C N N N N G T G G T G N N N N C A C
BstEII  (1184)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BseRI  (1186)
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  (1316)
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  (1316)
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.
BmgBI  (1318)
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.
PasI  (1348)
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.
StuI  (1517)
1 site		 A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (1548)
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.
BspDI  (1554)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1554)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (1613)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
SgrAI  (1700)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BssHII  (1925)
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.
PaeR7I  (1943)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1943)
1 site		 C T C G A G G A G C T C
AsiSI  (2001)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (2001)
1 site		 C G A T C G G C T A G C
SgfI  (2001)
1 site		 G C G A T C G C C G C T A G C G
DraI  (2368)
1 site		 T T T A A A A A A T T T
PmeI  (2368)
1 site		 G T T T A A A C C A A A T T T G
Acc65I  (2385)
1 site		 G G T A C C C C A T G G
KpnI  (2389)
1 site		 G G T A C C C C A T G G
XbaI  (2400)
1 site		 T C T A G A A G A T C T
SbfI  (2416)
1 site		 C C T G C A G G G G A C G T C C
BlpI  (2481)
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.
NotI  (2511)
1 site		 G C G G C C G C C G C C G G C G
HaloTag®
1067 .. 1957  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
1067 .. 1957  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
TEV site
1970 .. 1990  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
1970 .. 1990  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
AmpR
3006 .. 3866  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3006 .. 3074  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3006 .. 3866  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3075 .. 3866  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3006 .. 3866  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4024 .. 4612  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4024 .. 4612  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
barnase
2028 .. 2363  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
2028 .. 2363  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
cer region
4728 .. 5011  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
4728 .. 5011  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
SV40 poly(A) signal
2533 .. 2654  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2533 .. 2654  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2901 .. 3005  =  105 bp
AmpR promoter
2901 .. 3005  =  105 bp
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1067 .. 2014  =  948 bp
ORF:  315 amino acids  =  35.7 kDa
ORF:  2759 .. 3109  =  351 bp
ORF:  116 amino acids  =  13.1 kDa
ORF:  2028 .. 2363  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  3006 .. 3866  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1630 .. 1866  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  1845 .. 2084  =  240 bp
ORF:  79 amino acids  =  8.9 kDa
ORF:  821 .. 2029  =  1209 bp
ORF:  402 amino acids  =  44.8 kDa
ORF:  3470 .. 3736  =  267 bp
ORF:  88 amino acids  =  9.2 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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