pcDNA3.1+ N-HA

Mammalian vector with a CMV promoter for expressing N-terminally HA-tagged proteins.

Sequence Author: GenScript

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SgrDI (5448) SspI (5331) ScaI (5007) PvuI (4897) AhdI (4527) PciI (3634) BstZ17I (3255) BsmI (3203) PfoI (3058) BstBI (2965) RsrII (2799) BglII (12) MfeI (161) Bpu10I (180) NruI (208) MluI (228) SpeI (249) NdeI (484) SnaBI (590) NheI (895) BmtI (899) AflII (908) HindIII (911) HA Acc65I (953) KpnI (957) BamHI (965) EcoRI (971) EcoRV (983) BstXI (993) NotI (998) PaeR7I - PspXI - XhoI (1004) XbaI (1010) EcoO109I - PspOMI (1016) ApaI (1020) BbsI (1236) DraIII (1550) CsiI - SexAI * (1840) BseRI (2069) StuI (2072) AvrII (2073) TspMI - XmaI (2094) SmaI (2096) BsaBI * (2142) KasI (2282) NarI (2283) SfoI (2284) PluTI (2286) MscI (2365) PflFI - Tth111I (2401) BssHII (2680) pcDNA3.1+/N-HA 5450 bp
SgrDI  (5448)
1 site
C G T C G A C G G C A G C T G C
SspI  (5331)
1 site
A A T A T T T T A T A A
ScaI  (5007)
1 site
A G T A C T T C A T G A
PvuI  (4897)
1 site
C G A T C G G C T A G C
AhdI  (4527)
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  (3634)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3255)
1 site
G T A T A C C A T A T G
BsmI  (3203)
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.
PfoI  (3058)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstBI  (2965)
1 site
T T C G A A A A G C T T
RsrII  (2799)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
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
MluI  (228)
1 site
A C G C G T T G C G C A
SpeI  (249)
1 site
A C T A G T T G A T 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
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  (953)
1 site
G G T A C C C C A T G G
KpnI  (957)
1 site
G G T A C C C C A T G G
BamHI  (965)
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  (971)
1 site
G A A T T C C T T A A G
EcoRV  (983)
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  (993)
1 site
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  (998)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1004)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1004)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1004)
1 site
C T C G A G G A G C T C
XbaI  (1010)
1 site
T C T A G A A G A T C T
EcoO109I  (1016)
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  (1016)
1 site
G G G C C C C C C G G G
ApaI  (1020)
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  (1236)
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.
DraIII  (1550)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
CsiI  (1840)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (1840)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BseRI  (2069)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
StuI  (2072)
1 site
A G G C C T T C C G G A
AvrII  (2073)
1 site
C C T A G G G G A T C C
TspMI  (2094)
1 site
C C C G G G G G G C C C
XmaI  (2094)
1 site
C C C G G G G G G C C C </