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Plasmid Files

pcDNA™4/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.

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pcDNA4 HisMax-TOPO (linearized) Sequence and MappcDNA4 HisMax-TOPO (linearized).dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SgrDI (4088) SspI (3971) ScaI (3647) PvuI (3537) FspI (3389) AhdI (3167) PciI (2277) BspQI - SapI (2161) BstZ17I (1898) BsmI (1846) AanI (1785) FseI (1538) BmgBI (1396) SgrAI (1376) BglII (4102) MfeI (4251) Bpu10I (4270) NruI (4298) MluI (4318) SpeI (4339) 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) BstXI (27) EcoRI (31) PstI (40) EcoRV (43) BstXI (53) NotI (58) PaeR7I - PspXI - XhoI (64) XbaI (70) PspOMI (76) ApaI (80) BbsI (296) AanI (735) 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.
AanI  (1785)
2 sites
T T A T A A A A T A T T
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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

The sticky ends produced by AflII are hard to ligate.
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.
BstXI  (27)
2 sites
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.
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.
BstXI  (53)
2 sites
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.
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.
AanI  (735)
2 sites
T T A T A A A A T A T T
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
   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
   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
EM7 promoter
1196 .. 1243  =  48 bp
synthetic bacterial promoter
EM7 promoter
1196 .. 1243  =  48 bp
synthetic bacterial promoter
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
   Segment 1:  
   5239 .. 5247  =  9 bp
   3 amino acids  =  409.5 Da
Product: Xpress™ epitope tag, including an
enterokinase recognition and cleavage site
Xpress™ tag
5239 .. 5262  =  24 bp
8 amino acids  =  998.0 Da
   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
T7 promoter
4953 .. 4971  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4953 .. 4971  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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
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
SV40 ori
999 .. 1134  =  136 bp
SV40 origin of replication
SV40 ori
999 .. 1134  =  136 bp
SV40 origin of replication
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