pcDNA4 His C

Mammalian vector for expressing proteins with a cleavable N-terminal 6xHis tag. For other reading frames, use pcDNA™4/His A or pcDNA™4/His B.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (5094) SspI (4977) ScaI (4653) PvuI (4543) FspI (4395) AhdI (4173) PciI (3280) BspQI - SapI (3164) BstZ17I (2901) BsmI (2849) FseI (2541) BglII (12) MfeI (161) Bpu10I (180) NruI (208) MluI (228) SpeI (249) NdeI (484) SnaBI (590) CMV promoter Eco53kI (816) SacI (818) AflII (908) HindIII (911) ATG Acc65I (1012) KpnI (1016) BamHI (1019) EcoRI (1034) PstI (1043) EcoRV (1046) NotI (1061) PaeR7I - PspXI - XhoI (1067) XbaI (1073) EcoO109I - PspOMI (1079) ApaI (1083) bGH poly(A) signal BbsI (1299) StuI (2135) AvrII (2136) MscI (2268) BssHII - MauBI (2301) SgrAI (2379) BmgBI (2399) pcDNA™4/His C 5096 bp
SgrDI  (5094)
1 site
C G T C G A C G G C A G C T G C
SspI  (4977)
1 site
A A T A T T T T A T A A
ScaI  (4653)
1 site
A G T A C T T C A T G A
PvuI  (4543)
1 site
C G A T C G G C T A G C
FspI  (4395)
1 site
T G C G C A A C G C G T
AhdI  (4173)
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  (3280)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3164)
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  (3164)
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  (2901)
1 site
G T A T A C C A T A T G
BsmI  (2849)
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  (2541)
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
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
Eco53kI  (816)
1 site
G A G C T C C T C G A G
SacI  (818)
1 site
G A G C T C C T C G A 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  (1012)
1 site
G G T A C C C C A T G G
KpnI  (1016)
1 site
G G T A C C C C A T G G
BamHI  (1019)
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  (1034)
1 site
G A A T T C C T T A A G
PstI  (1043)
1 site
C T G C A G G A C G T C
EcoRV  (1046)
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  (1061)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1067)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1067)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1067)
1 site
C T C G A G G A G C T C
XbaI  (1073)
1 site
T C T A G A A G A T C T
EcoO109I  (1079)
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  (1079)
1 site
G G G C C C C C C G G G
ApaI  (1083)
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  (1299)
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  (2135)
1 site
A G G C C T T C C G G A
AvrII  (2136)
1 site
C C T A G G G G A T C C
MscI  (2268)
1 site
T G G C C A A C C G G T
BssHII  (2301)
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  (2301)
1 site
C G C G C G C G G C G C G C G C
SgrAI  (2379)
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  (2399)
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
4100 .. 4960  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4100 .. 4891  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4100 .. 4960  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4892 .. 4960  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4100 .. 4960  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3341 .. 3929  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3341 .. 3929  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1380 .. 1808  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1380