pcDNA4 HisMax-TOPO (linearized)

Linearized mammalian vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products and expression of N-terminally 6xHis-tagged proteins.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTOPO Cloning VectorspcDNA4 HisMax-TOPO (linearized)
SgrDI (4088) SspI (3971) ScaI (3647) PvuI (3537) FspI (3389) AhdI (3167) PciI (2277) BspQI - SapI (2161) BstZ17I (1898) BsmI (1846) FseI (1538) BmgBI (1396) SgrAI (1376) BglII (4102) MfeI (4251) Bpu10I (4270) NruI (4298) MluI (4318) SpeI (4339) CMV enhancer NdeI (4574) SnaBI (4680) Eco53kI (4906) SacI (4908) AflII (4998) HindIII (5001) BlpI (5004) SrfI (5051) ATG 6xHis T7 tag (gene 10 leader) Xpress™ tag End (5274) Start (1) BamHI (16) EcoRI (31) PstI (40) EcoRV (43) NotI (58) PaeR7I - PspXI - XhoI (64) XbaI (70) PspOMI (76) ApaI (80) BbsI (296) StuI (1132) MscI (1265) MauBI (1298) pcDNA™4/HisMax-TOPO® 5273 bp
SgrDI  (4088)
1 site		 C G T C G A C G G C A G C T G C
SspI  (3971)
1 site		 A A T A T T T T A T A A
ScaI  (3647)
1 site		 A G T A C T T C A T G A
PvuI  (3537)
1 site		 C G A T C G G C T A G C
FspI  (3389)
1 site		 T G C G C A A C G C G T
AhdI  (3167)
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.
PciI  (2277)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2161)
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  (2161)
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.
BstZ17I  (1898)
1 site		 G T A T A C C A T A T G
BsmI  (1846)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
FseI  (1538)
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.
BmgBI  (1396)
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.
SgrAI  (1376)
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.
BglII  (4102)
1 site		 A G A T C T T C T A G A
MfeI  (4251)
1 site		 C A A T T G G T T A A C
Bpu10I  (4270)
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.
NruI  (4298)
1 site		 T C G C G A A G C G C T
MluI  (4318)
1 site		 A C G C G T T G C G C A
SpeI  (4339)
1 site		 A C T A G T T G A T C A
NdeI  (4574)
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  (4680)
1 site		 T A C G T A A T G C A T
Eco53kI  (4906)
1 site		 G A G C T C C T C G A G
SacI  (4908)
1 site		 G A G C T C C T C G A G
AflII  (4998)
1 site		 C T T A A G G A A T T C
HindIII  (5001)
1 site		 A A G C T T T T C G A A
BlpI  (5004)
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.
SrfI  (5051)
1 site		 G C C C G G G C C G G G C C C G
End  (5274)
0 sites
Start  (1)
0 sites
BamHI  (16)
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  (31)
1 site		 G A A T T C C T T A A G
PstI  (40)
1 site		 C T G C A G G A C G T C
EcoRV  (43)
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.
NotI  (58)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (64)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (64)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (64)
1 site		 C T C G A G G A G C T C
XbaI  (70)
1 site		 T C T A G A A G A T C T
PspOMI  (76)
1 site		 G G G C C C C C C G G G
ApaI  (80)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BbsI  (296)
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.
StuI  (1132)
1 site		 A G G C C T T C C G G A
MscI  (1265)
1 site		 T G G C C A A C C G G T
MauBI  (1298)
1 site		 C G C G C G C G G C G C G C G C
AmpR
3094 .. 3954  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3094 .. 3885  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3094 .. 3954  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3886 .. 3954  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3094 .. 3954  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2338 .. 2923  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2338 .. 2923  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
377 .. 805  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
377 .. 805  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
4325 .. 4704  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4325 .. 4704  =  380 bp
human cytomegalovirus immediate early enhancer
BleoR
1262 .. 1636  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
1262 .. 1636  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
819 .. 1148  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
819 .. 1148  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
107 .. 331  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
107 .. 331  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
4705 .. 4908  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
4705 .. 4908  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
QBI SP163
5007 .. 5169  =  163 bp
strong translational enhancer derived from the 5' UTR of the mouse VEGF gene
QBI SP163
5007 .. 5169  =  163 bp
strong translational enhancer derived from the 5' UTR of the mouse VEGF gene
SV40 poly(A) signal
1766 .. 1887  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1766 .. 1887  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3955 .. 4059  =  105 bp
AmpR promoter
3955 .. 4059  =  105 bp
ATG
5170 .. 5172  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5170 .. 5172  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
5182 .. 5199  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5182 .. 5199  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 tag (gene 10 leader)
5203 .. 5235  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene 10
promotes efficient translation in E. coli
T7 tag (gene 10 leader)
5203 .. 5235  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene 10
promotes efficient translation in E. coli
Xpress™ tag
5239 .. 5262  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 1:  
   5239 .. 5247  =  9 bp
   3 amino acids  =  409.4 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
5239 .. 5262  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 2:  enterokinase site  
   5248 .. 5262  =  15 bp
   5 amino acids  =  606.5 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
5239 .. 5262  =  24 bp
8 amino acids  =  998.0 Da
2 segments
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
EM7 promoter
1196 .. 1243  =  48 bp
synthetic bacterial promoter
EM7 promoter
1196 .. 1243  =  48 bp
synthetic bacterial promoter
T7 promoter
4953 .. 4971  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4953 .. 4971  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
999 .. 1134  =  136 bp
SV40 origin of replication
SV40 ori
999 .. 1134  =  136 bp
SV40 origin of replication
ORF:  328 .. 594  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1262 .. 1636  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  3224 .. 3490  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1255 .. 1542  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  3094 .. 3954  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  5005 .. 5274  =  270 bp
ORF:  89 amino acids  =  9.6 kDa  (no start codon)
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Individual Sequences & Maps

pBAD-TOPO (linearized)
pcDNA5 FRT TO-TOPO (linearized)
pEF6 V5-His-TOPO (linearized)
pGlow-TOPO (linearized)
pBAD Thio-TOPO (linearized)
pcDNA5 FRT V5-His-TOPO (linearized)
pENTR D-TOPO
pIB V5-His-TOPO (linearized)
pBAD202 D-TOPO
pcDNA6.2 C-EmGFP-GW TOPO (linearized)
pENTR SD D-TOPO
pLenti6.3 V5-TOPO (linearized)
pBlue-TOPO (linearized)
pcDNA6.2 C-YFP-GW TOPO (linearized)
pENTR TEV D-TOPO
pLenti6 V5-D-TOPO
pcDNA3.1D V5-His-TOPO
pcDNA6.2 N-EmGFP-GW TOPO (linearized)
pENTR5'-TOPO (linearized)
pLenti7.3 V5-TOPO (linearized)
pcDNA3.2 V5 GW D-TOPO
pCR-XL-2-TOPO (linearized)
pET100 D-TOPO
pLPS-B (linearized)
pcDNA6.2 cGeneBLAzer-GW D-TOPO
pCR-XL-TOPO (linearized)
pET101 D-TOPO
pMT V5-His-TOPO (linearized)
pcDNA6.2 V5 GW D-TOPO
pCR2.1-TOPO (linearized)
pET102 D-TOPO
pOptiVEC-TOPO (linearized)
pcDNA3.1 CT-GFP-TOPO (linearized)
pCR4-TOPO (linearized)
pET151 D-TOPO
pSecTag FRT V5-His-TOPO (linearized)
pcDNA3.1 NT-GFP-TOPO (linearized)
pCR4Blunt-TOPO (linearized)
pET160 GW D-TOPO
pTOP Blunt V2 (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pCR8 GW TOPO (linearized)
pET161 GW D-TOPO
pTrcHis-TOPO (linearized)
pcDNA3.3-TOPO (linearized)
pCR-Blunt II-TOPO (linearized)
pET200 D-TOPO
pTrcHis2-TOPO (linearized)
pcDNA3.4-TOPO (linearized)
pCRII-TOPO (linearized)
pEXP5-CT TOPO (linearized)
pYES2.1 V5-His-TOPO (linearized)
pcDNA4 HisMax-TOPO (linearized)
pEF5 FRT V5-D-TOPO
pEXP5-NT TOPO (linearized)

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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