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Plasmid Files

pHet-Act1-2

Mammalian vector encoding a transcription factor activation domain and a DNA-binding domain that can be assembled by drug-induced heterodimerization.

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pHet-Act1-2 Sequence and MappHet-Act1-2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BglII (8197) AhdI (7297) BstBI (6003) Bpu10I (5984) BssHII (5843) SacII (5602) BstEII (5522) RsrII (5504) BsiWI (5444) PflFI - Tth111I (5430) SfiI (5275) NaeI (4535) NgoMIV (4533) MfeI (4231) BsrGI (96) SnaBI (493) Eco53kI (727) SacI (729) I-PpoI (851) NheI (1085) BmtI (1089) PaeR7I - XhoI (1091) ATG AleI (1414) MluI (1990) PmlI (2356) PflMI (2493) PciI (2530) BmgBI (2583) ATG BclI * (2976) HpaI (4222) pHet-Act1-2 8202 bp
BglII  (8197)
1 site
A G A T C T T C T A G A
AhdI  (7297)
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.
BstBI  (6003)
1 site
T T C G A A A A G C T T
Bpu10I  (5984)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
BssHII  (5843)
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.
SacII  (5602)
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.
BstEII  (5522)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII  (5504)
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.
BsiWI  (5444)
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.
PflFI  (5430)
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  (5430)
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.
SfiI  (5275)
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.
NaeI  (4535)
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  (4533)
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.
MfeI  (4231)
1 site
C A A T T G G T T A A C
BsrGI  (96)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SnaBI  (493)
1 site
T A C G T A A T G C A T
Eco53kI  (727)
1 site
G A G C T C C T C G A G
SacI  (729)
1 site
G A G C T C C T C G A G
I-PpoI  (851)
1 site
C T C T C T T A A G G T A G C G A G A G A A T T C C A T C G

I-PpoI is a homing endonuclease that can recognize a variety of
similar recognition sequences.
NheI  (1085)
1 site
G C T A G C C G A T C G
BmtI  (1089)
1 site
G C T A G C C G A T C G
PaeR7I  (1091)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1091)
1 site
C T C G A G G A G C T C
AleI  (1414)
1 site
C A C N N N N G T G G T G N N N N C A C
MluI  (1990)
1 site
A C G C G T T G C G C A
PmlI  (2356)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PflMI  (2493)
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.
PciI  (2530)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BmgBI  (2583)
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.
BclI  (2976)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
HpaI  (4222)
1 site
G T T A A C C A A T T G
AmpR
6510 .. 7370  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6510 .. 6578  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6510 .. 7370  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6579 .. 7370  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6510 .. 7370  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
5388 .. 5987  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
5388 .. 5987  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
7541 .. 8129  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7541 .. 8129  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
2040 .. 2613  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
2040 .. 2613  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
p65 activation domain
1399 .. 1971  =  573 bp
191 amino acids  =  19.4 kDa
Product: activation domain from the NFκB
transcription factor p65
p65 activation domain
1399 .. 1971  =  573 bp
191 amino acids  =  19.4 kDa
Product: activation domain from the NFκB
transcription factor p65
f1 ori
4408 .. 4863  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4408 .. 4863  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
ZFHD1
2689 .. 3051  =  363 bp
121 amino acids  =  14.5 kDa
   Segment 1:  Zif268  
   2689 .. 2862  =  174 bp
   58 amino acids  =  6.9 kDa
Product: composite human DNA-binding domain
composed of domains from the transcription factors
Zif268 and Oct-1 (Pomerantz et al., 1995)
ZFHD1
2689 .. 3051  =  363 bp
121 amino acids  =  14.5 kDa
   Segment 2:  
   2863 .. 2868  =  6 bp
   2 amino acids  =  132.1 Da
Product: composite human DNA-binding domain
composed of domains from the transcription factors
Zif268 and Oct-1 (Pomerantz et al., 1995)
ZFHD1
2689 .. 3051  =  363 bp
121 amino acids  =  14.5 kDa
   Segment 3:  Oct-1  
   2869 .. 3051  =  183 bp
   61 amino acids  =  7.5 kDa
Product: composite human DNA-binding domain
composed of domains from the transcription factors
Zif268 and Oct-1 (Pomerantz et al., 1995)
ZFHD1
2689 .. 3051  =  363 bp
121 amino acids  =  14.5 kDa
3 segments
Product: composite human DNA-binding domain
composed of domains from the transcription factors
Zif268 and Oct-1 (Pomerantz et al., 1995)
SV40 promoter
4980 .. 5337  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
4980 .. 5337  =  358 bp
SV40 enhancer and early promoter
FKBP (DmrA)
3058 .. 3378  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
3058 .. 3378  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
3712 .. 4032  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
3712 .. 4032  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
3388 .. 3705  =  318 bp
106 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
3388 .. 3705  =  318 bp
106 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FRB* (DmrC)
1114 .. 1392  =  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)
1114 .. 1392  =  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
518 .. 729  =  212 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
518 .. 729  =  212 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
890 .. 1022  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
890 .. 1022  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
4101 .. 4222  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4101 .. 4222  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6405 .. 6509  =  105 bp
AmpR promoter
6405 .. 6509  =  105 bp
poly(A) signal
6051 .. 6099  =  49 bp
synthetic polyadenylation signal
poly(A) signal
6051 .. 6099  =  49 bp
synthetic polyadenylation signal
c-myc NLS
2656 .. 2682  =  27 bp
9 amino acids  =  997.2 Da
Product: nuclear localization signal of human c-myc
(Dang and Lee, 1988)
c-myc NLS
2656 .. 2682  =  27 bp
9 amino acids  =  997.2 Da
Product: nuclear localization signal of human c-myc
(Dang and Lee, 1988)
ATG
1102 .. 1104  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1102 .. 1104  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2647 .. 2649  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2647 .. 2649  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
5188 .. 5323  =  136 bp
SV40 origin of replication
SV40 ori
5188 .. 5323  =  136 bp
SV40 origin of replication
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