pHet-Mem1

Mammalian vector encoding FKBP heterodimerizer domains, for creating membrane-associated fusion proteins that can be dimerized with a drug. Also known as pC4M-F2E.

Sequence Author: Clontech (TaKaRa)

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PvuII (5614) NaeI (5412) NgoMIV (5410) DraIII (5309) PfoI (4894) XmnI (4460) AhdI (3849) BspQI - SapI (2840) MluI (1) SnaBI (298) EcoRI (656) AfeI (707) XbaI (710) PaqCI (712) TspMI - XmaI (831) SmaI (833) FKBP (DmrA) EagI (1075) SphI (1129) FKBP (DmrA) SacII (1213) KasI (1289) NarI (1290) SfoI (1291) PluTI (1293) AleI (1331) BmgBI (1342) SpeI (1364) BsiWI (1374) BamHI (1409) KflI (1438) HincII (1590) MfeI (1780) PspOMI (1955) ApaI (1959) MscI (2100) BstXI (2108) BglII (2129) PflMI (2335) AbsI - PaeR7I - PspXI - XhoI (2578) AvrII (2601) StuI (2606) SfiI (2655) pHet-Mem1 5726 bp
PvuII  (5614)
1 site
C A G C T G G T C G A C
NaeI  (5412)
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  (5410)
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  (5309)
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  (4894)
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  (4460)
1 site
G A A N N N N T T C C T T N N N N A A G
AhdI  (3849)
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.
BspQI  (2840)
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  (2840)
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.
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
AfeI  (707)
1 site
A G C G C T T C G C G A
XbaI  (710)
1 site
T C T A G A A G A T C T
PaqCI  (712)
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  (831)
1 site
C C C G G G G G G C C C
XmaI  (831)
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  (833)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EagI  (1075)
1 site
C G G C C G G C C G G C
SphI  (1129)
1 site
G C A T G C C G T A C G
SacII  (1213)
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.
KasI  (1289)
1 site
G G C G C C C C G C G G
NarI  (1290)
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  (1291)
1 site
G G C G C C C C G C G G
PluTI  (1293)
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.
AleI  (1331)
1 site
C A C N N N N G T G G T G N N N N C A C
BmgBI  (1342)
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.
SpeI  (1364)
1 site
A C T A G T T G A T C A
BsiWI  (1374)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BamHI  (1409)
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  (1438)
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  (1590)
1 site
G T Y R A C C A R Y T G
MfeI  (1780)
1 site
C A A T T G G T T A A C
PspOMI  (1955)
1 site
G G G C C C C C C G G G
ApaI  (1959)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
MscI  (2100)
1 site
T G G C C A A C C G G T
BstXI  (2108)
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  (2129)
1 site
A G A T C T T C T A G A
PflMI  (2335)
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  (2578)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2578)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2578)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2578)
1 site
C T C G A G G A G C T C
AvrII  (2601)
1 site
C C T A G G G G A T C C
StuI  (2606)
1 site
A G G C C T T C C G G A
SfiI  (2655)
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.
AmpR
3776 .. 4648  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
   Segment 2:  
   3776 .. 4579  =  804 bp
   267 amino acids  =  29.4 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3776 .. 4648  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
   Segment 1:  signal sequence  
   4580 .. 4648  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3776 .. 4648  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
N-myristoylation signal
668 .. 709  =  42 bp
14 amino acids  =  1.5 kDa
Product: N-myristoylation signal from Src kinase
N-myristoylation signal
668 .. 709  =  42 bp
14 amino acids  =  1.5 kDa
Product: N-myristoylation signal from Src kinase
FKBP (DmrA)
716 .. 1036  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
716 .. 1036  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
1043 .. 1363  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
synthetic gene with alternative codons
FKBP (DmrA)
1043 .. 1363  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
synthetic gene with alternative codons
HA
1370 .. 1396  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
1370 .. 1396  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
ori
3017 .. 3605  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3017 .. 3605  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
β-globin intron
1428 .. 2000  =  573 bp
intron from rabbit β-globin gene
β-globin intron
1428 .. 2000  =  573 bp
intron from rabbit β-globin gene
f1 ori
5085 .. 5540  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5085 .. 5540  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
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
2585 .. 2780  =  196 bp
SV40 promoter
2585 .. 2780  =  196 bp
AmpR promoter
4649 .. 4753  =  105 bp
AmpR promoter
4649 .. 4753  =  105 bp
β-globin poly(A) signal
2197 .. 2252  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
2197 .. 2252  =  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
2605 .. 2740  =  136 bp
SV40 origin of replication
SV40 ori
2605 .. 2740  =  136 bp
SV40 origin of replication
ORF:  668 .. 1399  =  732 bp
ORF:  243 amino acids  =  26.9 kDa
ORF:  3906 .. 4172  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1025 .. 1480  =  456 bp
ORF:  151 amino acids  =  15.6 kDa
ORF:  3776 .. 4648  =  873 bp
ORF:  290 amino acids  =  32.0 kDa
ORF:  5556 .. 105  =  276 bp
ORF:  91 amino acids  =  9.5 kDa
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