pcDNA4 V5-His A

Mammalian vector for expressing C-terminally V5- and 6xHis-tagged proteins. For other reading frames, use pcDNA™4/V5-His B or pcDNA™4/V5-His C.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (5090) SspI (4973) ScaI (4649) PvuI (4539) FspI (4391) AhdI (4169) PciI (3276) BspQI - SapI (3160) BstZ17I (2897) BsmI (2845) FseI (2537) BglII (12) MfeI (161) Bpu10I (180) NruI (208) NdeI (484) SnaBI (590) NheI (895) BmtI (899) AflII (908) HindIII (911) Acc65I (917) KpnI (921) BamHI (929) EcoRI (952) PstI (961) EcoRV (964) NotI (979) PaeR7I - PspXI - XhoI (985) XbaI (991) PspOMI (997) ApaI (1001) BstBI (1004) AgeI (1054) BbsI (1295) StuI (2131) AvrII (2132) MscI (2264) BssHII - MauBI (2297) SgrAI (2375) BmgBI (2395) pcDNA™4/V5-His A 5092 bp
SgrDI  (5090)
1 site
C G T C G A C G G C A G C T G C
SspI  (4973)
1 site
A A T A T T T T A T A A
ScaI  (4649)
1 site
A G T A C T T C A T G A
PvuI  (4539)
1 site
C G A T C G G C T A G C
FspI  (4391)
1 site
T G C G C A A C G C G T
AhdI  (4169)
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  (3276)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3160)
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  (3160)
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  (2897)
1 site
G T A T A C C A T A T G
BsmI  (2845)
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  (2537)
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.
BglII  (12)
1 site
A G A T C T T C T A G A
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
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
NheI  (895)
1 site
G C T A G C C G A T C G
BmtI  (899)
1 site
G C T A G C C G A T C G
AflII  (908)
1 site
C T T A A G G A A T T C
HindIII  (911)
1 site
A A G C T T T T C G A A
Acc65I  (917)
1 site
G G T A C C C C A T G G
KpnI  (921)
1 site
G G T A C C C C A T G G
BamHI  (929)
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  (952)
1 site
G A A T T C C T T A A G
PstI  (961)
1 site
C T G C A G G A C G T C
EcoRV  (964)
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  (979)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (985)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (985)
1 site
V C T C G A G B B G A G C T C V
XhoI  (985)
1 site
C T C G A G G A G C T C
XbaI  (991)
1 site
T C T A G A A G A T C T
PspOMI  (997)
1 site
G G G C C C C C C G G G
ApaI  (1001)
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  (1004)
1 site
T T C G A A A A G C T T
AgeI  (1054)
1 site
A C C G G T T G G C C A
BbsI  (1295)
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  (2131)
1 site
A G G C C T T C C G G A
AvrII  (2132)
1 site
C C T A G G G G A T C C
MscI  (2264)
1 site
T G G C C A A C C G G T
BssHII  (2297)
1 site
G C G C G C