pcDNA4 myc-His B

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

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (5077) SspI (4960) ScaI (4636) PvuI (4526) FspI (4378) AhdI (4156) PciI (3263) BspQI - SapI (3147) BstZ17I (2884) BsmI (2832) FseI (2524) BglII (12) MfeI (161) Bpu10I (180) NruI (208) MluI (228) NdeI (484) SnaBI (590) HindIII (902) Acc65I (908) KpnI (912) BamHI (920) EcoRI (943) PstI (952) 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) StuI (2118) AvrII (2119) MscI (2251) BssHII - MauBI (2284) SgrAI (2362) BmgBI (2382) pcDNA™4/myc-His B 5079 bp
SgrDI  (5077)
1 site
C G T C G A C G G C A G C T G C
SspI  (4960)
1 site
A A T A T T T T A T A A
ScaI  (4636)
1 site
A G T A C T T C A T G A
PvuI  (4526)
1 site
C G A T C G G C T A G C
FspI  (4378)
1 site
T G C G C A A C G C G T
AhdI  (4156)
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  (3263)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3147)
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  (3147)
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  (2884)
1 site
G T A T A C C A T A T G
BsmI  (2832)
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  (2524)
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
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
PstI  (952)
1 site
C T G C A G G A C G T C
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.
StuI  (2118)
1 site
A G G C C T T C C G G A
AvrII  (2119)
1 site
C C T A G G G G A T C C
MscI  (2251)
1 site
T G G C C A A C C G G T
BssHII  (2284)
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  (2284)
1 site
C G C G C G C G G C G C G C G C
SgrAI  (2362)
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  (2382)
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
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4083 .. 4874  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4875 .. 4943  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3324 .. 3912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3324 .. 3912  =  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
BleoR
2248 .. 2622  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
2248 .. 2622  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
1804 .. 2134  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
1804 .. 2134  =  331 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
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
2752 .. 2873  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2752 .. 2873  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4944 .. 5048  =  105 bp
AmpR promoter
4944 .. 5048  =  105 bp
MCS
902 .. 1003  =  102 bp
multiple cloning site
MCS
902 .. 1003  =  102 bp
multiple cloning site
EM7 promoter
2182 .. 2229  =  48 bp
synthetic bacterial promoter
EM7 promoter
2182 .. 2229  =  48 bp
synthetic bacterial promoter
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
1985 .. 2120  =  136 bp
SV40 origin of replication
SV40 ori
1985 .. 2120  =  136 bp
SV40 origin of replication
Myc
1001 .. 1030  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
1001 .. 1030  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
6xHis
1046 .. 1063  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1046 .. 1063  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag