pHet-Nuc1

Mammalian vector encoding an FKBP heterodimerizer domain, for creating soluble fusion proteins that can be dimerized with a drug. Also known as pC4EN-F1.

Sequence Author: Clontech (TaKaRa)

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PvuII (5298) NaeI (5096) NgoMIV (5094) DraIII (4993) PfoI (4578) XmnI (4144) BsaI (3594) AhdI (3533) MluI (1) CMV enhancer SnaBI (298) EcoRI (656) ATG BmgBI (696) HA SV40 NLS XbaI (754) PaqCI (756) TspMI - XmaI (875) SmaI (877) FKBP (DmrA) SpeI (1081) BamHI (1093) KflI (1122) HincII (1274) MfeI (1464) PspOMI (1639) ApaI (1643) Bsu36I (1743) MscI (1784) BstXI (1792) BglII (1813) PflMI (2019) AbsI - PaeR7I - PspXI - XhoI (2262) AvrII (2285) StuI (2290) SfiI (2339) BspQI - SapI (2524) pHet-Nuc1 5410 bp
PvuII  (5298)
1 site
C A G C T G G T C G A C
NaeI  (5096)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (5094)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
DraIII  (4993)
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.
PfoI  (4578)
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.
XmnI  (4144)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaI  (3594)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (3533)
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.
MluI  (1)
1 site
A C G C G T T G C G C A
SnaBI  (298)
1 site
T A C G T A A T G C A T
EcoRI  (656)
1 site
G A A T T C C T T A A G
BmgBI  (696)
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.
XbaI  (754)
1 site
T C T A G A A G A T C T
PaqCI  (756)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
TspMI  (875)
1 site
C C C G G G G G G C C C
XmaI  (875)
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  (877)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SpeI  (1081)
1 site
A C T A G T T G A T C A
BamHI  (1093)
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.
KflI  (1122)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
HincII  (1274)
1 site
G T Y R A C C A R Y T G
MfeI  (1464)
1 site
C A A T T G G T T A A C
PspOMI  (1639)
1 site
G G G C C C C C C G G G
ApaI  (1643)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
Bsu36I  (1743)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
MscI  (1784)
1 site
T G G C C A A C C G G T
BstXI  (1792)
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.
BglII  (1813)
1 site
A G A T C T T C T A G A
PflMI  (2019)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
AbsI  (2262)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2262)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2262)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2262)
1 site
C T C G A G G A G C T C
AvrII  (2285)
1 site
C C T A G G G G A T C C
StuI  (2290)
1 site
A G G C C T T C C G G A
SfiI  (2339)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BspQI  (2524)
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  (2524)
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.
AmpR
3460 .. 4332  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
   Segment 2:  
   3460 .. 4263  =  804 bp
   267 amino acids  =  29.4 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3460 .. 4332  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
   Segment 1:  signal sequence  
   4264 .. 4332  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3460 .. 4332  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2701 .. 3289  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2701 .. 3289  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
β-globin intron
1112 .. 1684  =  573 bp
intron from rabbit β-globin gene
β-globin intron
1112 .. 1684  =  573 bp
intron from rabbit β-globin gene
f1 ori
4769 .. 5224  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4769 .. 5224  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
ATG
673 .. 675  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
673 .. 675  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
HA
685 .. 711  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
685 .. 711  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
SV40 NLS
733 .. 753  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
733 .. 753  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
FKBP (DmrA)
760 .. 1080  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
760 .. 1080  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
CMV enhancer
19 .. 322  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
19 .. 322  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
323 .. 526  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
323 .. 526  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 promoter
2269 .. 2464  =  196 bp
SV40 promoter
2269 .. 2464  =  196 bp
AmpR promoter
4333 .. 4437  =  105 bp
AmpR promoter
4333 .. 4437  =  105 bp
β-globin poly(A) signal
1881 .. 1936  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
1881 .. 1936  =  56 bp
rabbit β-globin polyadenylation signal
HSV TK 5'-UTR
616 .. 655  =  40 bp
5' untranslated region from the herpes simplex virus thymidine kinase gene
HSV TK 5'-UTR
616 .. 655  =  40 bp
5' untranslated region from the herpes simplex virus thymidine kinase gene
SV40 ori
2289 .. 2424  =  136 bp
SV40 origin of replication
SV40 ori
2289 .. 2424  =  136 bp
SV40 origin of replication
ORF:  673 .. 1092  =  420 bp
ORF:  139 amino acids  =  15.3 kDa
ORF:  3590 .. 3856  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5240 .. 105  =  276 bp
ORF:  91 amino acids  =  9.5 kDa
ORF:  3460 .. 4332  =  873 bp
ORF:  290 amino acids  =  32.0 kDa
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