pcDNA4 TO myc-His B

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

Sequence Author: Thermo Fisher (Invitrogen)

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SspI (5036) ScaI (4712) PvuI (4602) FspI (4454) BsaI (4293) AhdI (4232) PciI (3339) BspQI - SapI (3223) BstZ17I (2960) BsmI (2908) FseI (2600) 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) EcoO109I - PspOMI (1064) ApaI (1068) SacII (1071) BstBI (1075) AgeI (1116) StuI (2194) AvrII (2195) MscI (2327) BssHII - MauBI (2360) SgrAI (2438) BmgBI (2458) pcDNA™4/TO/myc-His B 5155 bp
SspI  (5036)
1 site
A A T A T T T T A T A A
ScaI  (4712)
1 site
A G T A C T T C A T G A
PvuI  (4602)
1 site
C G A T C G G C T A G C
FspI  (4454)
1 site
T G C G C A A C G C G T
BsaI  (4293)
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  (4232)
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  (3339)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3223)
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  (3223)
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  (2960)
1 site
G T A T A C C A T A T G
BsmI  (2908)
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  (2600)
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
EcoO109I  (1064)
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.
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.
SacII  (1071)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BstBI  (1075)
1 site
T T C G A A A A G C T T
AgeI  (1116)
1 site
A C C G G T T G G C C A
StuI  (2194)
1 site
A G G C C T T C C G G A
AvrII  (2195)
1 site
C C T A G G G G A T C C
MscI  (2327)
1 site
T G G C C A A C C G G T
BssHII  (2360)
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  (2360)
1 site
C G C G C G C G G C G C G C G C
SgrAI  (2438)
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  (2458)
1 site
C A C G T C G