pcDNA4 TO myc-His A

Vector for tetracycline-inducible expression of C-terminally Myc- and 6xHis-tagged proteins. For other reading frames, use pcDNA™4/TO/myc-His B or pcDNA™4/TO/myc-His C.

Sequence Author: Thermo Fisher (Invitrogen)

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SspI (5032) ScaI (4708) PvuI (4598) FspI (4450) BsaI (4289) AhdI (4228) PciI (3335) BspQI - SapI (3219) BstZ17I (2956) BsmI (2904) FseI (2596) MfeI (161) Bpu10I (180) NruI (208) MluI (228) NdeI (484) SnaBI (590) tet operator tet operator BsmBI - Esp3I (893) BspEI (955) AflII (975) HindIII (978) Acc65I (984) KpnI (988) BamHI (996) EcoRI (1019) PstI (1028) EcoRV (1031) NotI (1046) PaeR7I - PspXI - XhoI (1052) XbaI (1058) PspOMI (1064) ApaI (1068) BstBI (1071) AgeI (1112) StuI (2190) AvrII (2191) MscI (2323) BssHII - MauBI (2356) SgrAI (2434) BmgBI (2454) pcDNA™4/TO/myc-His A 5151 bp
SspI  (5032)
1 site
A A T A T T T T A T A A
ScaI  (4708)
1 site
A G T A C T T C A T G A
PvuI  (4598)
1 site
C G A T C G G C T A G C
FspI  (4450)
1 site
T G C G C A A C G C G T
BsaI  (4289)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (4228)
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  (3335)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3219)
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  (3219)
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  (2956)
1 site
G T A T A C C A T A T G
BsmI  (2904)
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  (2596)
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.
MfeI  (161)
1 site
C A A T T G G T T A A C
Bpu10I  (180)
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  (208)
1 site
T C G C G A A G C G C T
MluI  (228)
1 site
A C G C G T T G C G 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
BsmBI  (893)
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  (893)
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.
BspEI  (955)
1 site
T C C G G A A G G C C T
AflII  (975)
1 site
C T T A A G G A A T T C
HindIII  (978)
1 site
A A G C T T T T C G A A
Acc65I  (984)
1 site
G G T A C C C C A T G G
KpnI  (988)
1 site
G G T A C C C C A T G G
BamHI  (996)
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  (1019)
1 site
G A A T T C C T T A A G
PstI  (1028)
1 site
C T G C A G G A C G T C
EcoRV  (1031)
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  (1046)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1052)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1052)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1052)
1 site
C T C G A G G A G C T C
XbaI  (1058)
1 site
T C T A G A A G A T C T
PspOMI  (1064)
1 site
G G G C C C C C C G G G
ApaI  (1068)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BstBI  (1071)
1 site
T T C G A A A A G C T T
AgeI  (1112)
1 site
A C C G G T T G G C C A
StuI  (2190)
1 site
A G G C C T T C C G G A
AvrII  (2191)
1 site
C C T A G G G G A T C C
MscI  (2323)
1 site
T G G C C A A C C G G T
BssHII  (2356)
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.
MauBI  (2356)
1 site
C G C G C G C G G C G C G C G C
SgrAI  (2434)
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.
BmgBI  (2454)
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.
AmpR
4155 .. 5015  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4155 .. 4946  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4155 .. 5015  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4947 .. 5015  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4155 .. 5015  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3396 .. 3984  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3396 .. 3984  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1434 .. 1862  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1434 .. 1862  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
BleoR
2320 .. 2694  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
2320 .. 2694  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
1876 .. 2206  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
1876 .. 2206  =  331 bp
SV40 enhancer and early promoter
bGH poly(A) signal
1164 .. 1388  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1164 .. 1388  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
2824 .. 2945  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2824 .. 2945  =  122 bp
SV40 polyadenylation signal
AmpR promoter
5016 .. 5120  =  105 bp
AmpR promoter
5016 .. 5120  =  105 bp
MCS
968 .. 1069  =  102 bp
multiple cloning site
MCS
968 .. 1069  =  102 bp
multiple cloning site
Myc
1073 .. 1102  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
1073 .. 1102  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
6xHis
1118 .. 1135  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1118 .. 1135  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
EM7 promoter
2254 .. 2301  =  48 bp
synthetic bacterial promoter
EM7 promoter
2254 .. 2301  =  48 bp
synthetic bacterial promoter
tet operator
820 .. 838  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
820 .. 838  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
841 .. 859  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
841 .. 859  =  19 bp
bacterial operator O2 for the tetR and tetA genes
SV40 ori
2057 .. 2192  =  136 bp
SV40 origin of replication
SV40 ori
2057 .. 2192  =  136 bp
SV40 origin of replication
TATA box
804 .. 810  =  7 bp
TATA box
804 .. 810  =  7 bp
ORF:  2320 .. 2694  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  4285 .. 4551  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1385 .. 1651  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  774 .. 1103  =  330 bp
ORF:  109 amino acids  =  12.8 kDa
ORF:  2313 .. 2600  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  4155 .. 5015  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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