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Plasmid Files

pHet-1

Mammalian vector encoding an FRB heterodimerizer domain, for creating soluble fusion proteins that can be dimerized with a drug. Also known as pC4-RHE.

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pHet-1 Sequence and MappHet-1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 PvuII (5217) NaeI (5015) NgoMIV (5013) DraIII (4912) PfoI (4497) XmnI (4063) BsaI (3513) AhdI (3452) PspFI (2867) BseYI (2863) MluI (1) CMV enhancer SnaBI (298) BbsI (592) EcoRI (656) ATG XbaI (682) EcoNI (721) SphI (793) FRB* (DmrC) SpeI (967) BsiWI (977) HA BamHI (1012) KflI (1041) HincII (1193) MfeI (1383) Bsu36I (1662) MscI (1703) BstXI (1711) BglII (1732) PflMI (1938) AbsI - AvaI - BsoBI - PaeR7I - PspXI - XhoI (2181) BmeT110I (2182) AvrII (2204) SfiI (2258) BspQI - SapI (2443) pHet-1 5329 bp
PvuII  (5217)
1 site
C A G C T G G T C G A C
NaeI  (5015)
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  (5013)
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  (4912)
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  (4497)
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  (4063)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaI  (3513)
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  (3452)
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.
PspFI  (2867)
1 site
C C C A G C G G G T C G
BseYI  (2863)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
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
BbsI  (592)
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.
EcoRI  (656)
1 site
G A A T T C C T T A A G
XbaI  (682)
1 site
T C T A G A A G A T C T
EcoNI  (721)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SphI  (793)
1 site
G C A T G C C G T A C G
SpeI  (967)
1 site
A C T A G T T G A T C A
BsiWI  (977)
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  (1012)
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  (1041)
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  (1193)
1 site
G T Y R A C C A R Y T G
MfeI  (1383)
1 site
C A A T T G G T T A A C
Bsu36I  (1662)
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  (1703)
1 site
T G G C C A A C C G G T
BstXI  (1711)
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  (1732)
1 site
A G A T C T T C T A G A
PflMI  (1938)
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  (2181)
1 site
C C T C G A G G G G A G C T C C
AvaI  (2181)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2181)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
PaeR7I  (2181)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2181)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2181)
1 site
C T C G A G G A G C T C
BmeT110I  (2182)
1 site
C Y C G R G G R G C Y C
AvrII  (2204)
1 site
C C T A G G G G A T C C
SfiI  (2258)
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  (2443)
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  (2443)
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
3379 .. 4251  =  873 bp
290 amino acids  =  32.0 kDa
   Segment 2:  
   3379 .. 4182  =  804 bp
   267 amino acids  =  29.4 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3379 .. 4251  =  873 bp
290 amino acids  =  32.0 kDa
   Segment 1:  signal sequence  
   4183 .. 4251  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3379 .. 4251  =  873 bp
290 amino acids  =  32.0 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2620 .. 3208  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2620 .. 3208  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
β-globin intron
1031 .. 1603  =  573 bp
intron from rabbit β-globin gene
β-globin intron
1031 .. 1603  =  573 bp
intron from rabbit β-globin gene
f1 ori
4688 .. 5143  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4688 .. 5143  =  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
FRB* (DmrC)
688 .. 966  =  279 bp
93 amino acids  =  11.3 kDa
Product: T2098L mutant of FKBP-rapamycin binding
domain of human FRAP
binds synthetic ligands such as AP21967
FRB* (DmrC)
688 .. 966  =  279 bp
93 amino acids  =  11.3 kDa
Product: T2098L mutant of FKBP-rapamycin binding
domain of human FRAP
binds synthetic ligands such as AP21967
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
2188 .. 2383  =  196 bp
SV40 promoter
2188 .. 2383  =  196 bp
AmpR promoter
4252 .. 4356  =  105 bp
AmpR promoter
4252 .. 4356  =  105 bp
β-globin poly(A) signal
1800 .. 1855  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
1800 .. 1855  =  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
HA
973 .. 999  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
HA
973 .. 999  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
ATG
676 .. 678  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
676 .. 678  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
2208 .. 2343  =  136 bp
SV40 origin of replication
SV40 ori
2208 .. 2343  =  136 bp
SV40 origin of replication
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