pcDNA3.1 myc-His B

Mammalian vector for expressing C-terminally Myc- and 6xHis-tagged proteins. For other reading frames, use pcDNA™3.1/myc-His A or pcDNA™3.1/myc-His C.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (5495) SspI (5378) ScaI (5054) PvuI (4944) AhdI (4574) PciI (3681) BstZ17I (3302) BsmI (3250) PfoI (3105) RsrII (2844) BglII (12) MfeI (161) Bpu10I (180) MluI (228) NdeI (484) SnaBI (590) HindIII (902) Acc65I (908) KpnI (912) BamHI (920) EcoRI (943) EcoRV (955) NotI (970) PaeR7I - PspXI - XhoI (976) XbaI (982) EcoO109I - PspOMI (988) ApaI (992) SacII (995) BstBI (999) AgeI (1040) PmeI (1070) BbsI (1281) DraIII (1595) CsiI - SexAI * (1885) BseRI (2114) StuI (2117) AvrII (2118) TspMI - XmaI (2139) SmaI (2141) BsaBI * (2187) KasI (2327) NarI (2328) SfoI (2329) PluTI (2331) MscI (2410) PflFI - Tth111I (2446) BssHII (2725) pcDNA™3.1/myc-His B 5497 bp
SgrDI  (5495)
1 site
C G T C G A C G G C A G C T G C
SspI  (5378)
1 site
A A T A T T T T A T A A
ScaI  (5054)
1 site
A G T A C T T C A T G A
PvuI  (4944)
1 site
C G A T C G G C T A G C
AhdI  (4574)
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  (3681)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3302)
1 site
G T A T A C C A T A T G
BsmI  (3250)
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  (3105)
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.
RsrII  (2844)
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.
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
HindIII  (902)
1 site
A A G C T T T T C G A A
Acc65I  (908)
1 site
G G T A C C C C A T G G
KpnI  (912)
1 site
G G T A C C C C A T G G
BamHI  (920)
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  (943)
1 site
G A A T T C C T T A A G
EcoRV  (955)
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  (970)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (976)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (976)
1 site
V C T C G A G B B G A G C T C V
XhoI  (976)
1 site
C T C G A G G A G C T C
XbaI  (982)
1 site
T C T A G A A G A T C T
EcoO109I  (988)
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  (988)
1 site
G G G C C C C C C G G G
ApaI  (992)
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  (995)
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  (999)
1 site
T T C G A A A A G C T T
AgeI  (1040)
1 site
A C C G G T T G G C C A
PmeI  (1070)
1 site
G T T T A A A C C A A A T T T G
BbsI  (1281)
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  (1595)
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  (1885)
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  (1885)
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  (2114)
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  (2117)
1 site
A G G C C T T C C G G A
AvrII  (2118)
1 site
C C T A G G G G A T C C
TspMI  (2139)
1 site
C C C G G G G G G C C C
XmaI  (2139)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2141)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BsaBI  (2187)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
KasI  (2327)
1 site
G G C G C C C C G C G G
NarI  (2328)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2329)
1 site
G G C G C C C C G C G G
PluTI  (2331)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
MscI  (2410)
1 site
T G G C C A A C C G G T
PflFI  (2446)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2446)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BssHII  (2725)
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.
AmpR
4501 .. 5361  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4501 .. 5292  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4501 .. 5361  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5293 .. 5361  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4501 .. 5361  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2200 .. 2994  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2200 .. 2994  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
3742 .. 4330  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3742 .. 4330  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1362 .. 1790  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1362 .. 1790  =  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
SV40 promoter
1804 .. 2133  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1804 .. 2133  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
1092 .. 1316  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1092 .. 1316  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter