pZFHD1-2

Mammalian vector encoding a promoter that can be activated by drug-induced heterodimerization of ZFHD1 and the p65 activation domain.

Sequence Author: Clontech (TaKaRa)

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AfeI (14) BsgI (3930) PaqCI (3902) ScaI (3837) TatI (3835) ApaLI (3459) BsaI (2844) PfoI (2630) RsrII (2371) BsrDI (2088) PflFI - Tth111I (1973) BglII (27) Eco53kI (36) SacI (38) Acc65I (63) KpnI (67) SacII (70) MluI (76) NheI (82) BmtI (86) AseI (311) XmnI (312) minimal IL-2 promoter AgeI - SgrAI (462) PspOMI (468) ApaI (472) TspMI - XmaI (474) SmaI (476) BamHI (480) SpeI (486) BclI * (492) NotI (499) MfeI (605) HpaI (618) BtsI - BtsαI (694) AflII (737) DraIII (971) CsiI - SexAI * (1444) BglI - SfiI (1630) BseRI (1673) StuI (1676) KasI (1854) NarI (1855) SfoI (1856) PluTI (1858) MscI (1937) FspI (1957) pZFHD1-2 3991 bp
AfeI  (14)
1 site
A G C G C T T C G C G A
BsgI  (3930)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PaqCI  (3902)
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.
ScaI  (3837)
1 site
A G T A C T T C A T G A
TatI  (3835)
1 site
W G T A C W W C A T G W
ApaLI  (3459)
1 site
G T G C A C C A C G T G
BsaI  (2844)
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.
PfoI  (2630)
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.
RsrII  (2371)
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.
BsrDI  (2088)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (1973)
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  (1973)
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.
BglII  (27)
1 site
A G A T C T T C T A G A
Eco53kI  (36)
1 site
G A G C T C C T C G A G
SacI  (38)
1 site
G A G C T C C T C G A G
Acc65I  (63)
1 site
G G T A C C C C A T G G
KpnI  (67)
1 site
G G T A C C C C A T G G
SacII  (70)
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.
MluI  (76)
1 site
A C G C G T T G C G C A
NheI  (82)
1 site
G C T A G C C G A T C G
BmtI  (86)
1 site
G C T A G C C G A T C G
AseI  (311)
1 site
A T T A A T T A A T T A
XmnI  (312)
1 site
G A A N N N N T T C C T T N N N N A A G
AgeI  (462)
1 site
A C C G G T T G G C C A
SgrAI  (462)
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.
PspOMI  (468)
1 site
G G G C C C C C C G G G
ApaI  (472)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
TspMI  (474)
1 site
C C C G G G G G G C C C
XmaI  (474)
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  (476)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (480)
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.
SpeI  (486)
1 site
A C T A G T T G A T C A
BclI  (492)
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.
NotI  (499)
1 site
G C G G C C G C C G C C G G C G
MfeI  (605)
1 site
C A A T T G G T T A A C
HpaI  (618)
1 site
G T T A A C C A A T T G
BtsI  (694)
1 site
G C A G T G N N C G T C A C
BtsαI  (694)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
AflII  (737)
1 site
C T T A A G G A A T T C
DraIII  (971)
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.
CsiI  (1444)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (1444)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BglI  (1630)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SfiI  (1630)
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.
BseRI  (1673)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
StuI  (1676)
1 site
A G G C C T T C C G G A
KasI  (1854)
1 site
G G C G C C C C G C G G
NarI  (1855)
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  (1856)
1 site
G G C G C C C C G C G G
PluTI  (1858)
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.
MscI  (1937)
1 site
T G G C C A A C C G G T
FspI  (1957)
1 site
T G C G C A A C G C G T
NeoR/KanR
1727 .. 2521  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1727 .. 2521  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3129 .. 3717  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3129 .. 3717  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
747 .. 1202  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
747 .. 1202  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1335 .. 1692  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1335 .. 1692  =  358 bp
SV40 enhancer and early promoter
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 1:  
   88 .. 99  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 2:  
   100 .. 105  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 3:  
   106 .. 117  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 4:  
   118 .. 123  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 5:  
   124 .. 135  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 6:  
   136 .. 141  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 7:  
   142 .. 153  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 8:  
   154 .. 159  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 9:  
   160 .. 171  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 10:  
   172 .. 177  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 11:  
   178 .. 189  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 12:  
   190 .. 195  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 13:  
   196 .. 207  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 14:  
   208 .. 213  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 15:  
   214 .. 225  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 16:  
   226 .. 231  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 17:  
   232 .. 243  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 18:  
   244 .. 249  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 19:  
   250 .. 261  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 20:  
   262 .. 267  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 21:  
   268 .. 279  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 22:  
   280 .. 285  =  6 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
   Segment 23:  
   286 .. 297  =  12 bp
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
ZFHD1 binding sites
88 .. 297  =  210 bp
23 segments
12 binding sites for the composite human DNA-binding domain ZFHD1 (Pomerantz et al., 1995)
SV40 poly(A) signal
619 .. 740  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
619 .. 740  =  122 bp
SV40 polyadenylation signal
minimal IL-2 promoter
320 .. 433  =  114 bp
minimal promoter from the human interleukin-2 gene
minimal IL-2 promoter
320 .. 433  =  114 bp
minimal promoter from the human interleukin-2 gene
AmpR promoter
1229 .. 1333  =  105 bp
AmpR promoter
1229 .. 1333  =  105 bp
pause site
3841 .. 3932  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
3841 .. 3932  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
poly(A) signal
3779 .. 3827  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3779 .. 3827  =  49 bp
synthetic polyadenylation signal
HSV TK poly(A) signal
2753 .. 2800  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
2753 .. 2800  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
MCS
462 .. 505  =  44 bp
multiple cloning site
MCS
462 .. 505  =  44 bp
multiple cloning site
SV40 ori
1543 .. 1678  =  136 bp
SV40 origin of replication
SV40 ori
1543 .. 1678  =  136 bp
SV40 origin of replication
ORF:  2542 .. 2991  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  1727 .. 2521  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1899 .. 2285  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2036 .. 2572  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  2747 .. 2980  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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