pPTunerC

Vector for fusing a destabilization domain to the C-terminus of a partner protein.

Sequence Author: Clontech (TaKaRa)

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AflIII - PciI (4273) EcoO109I (3453) BsaI (3344) PfoI (3130) RsrII (2871) BsrDI (2588) PflFI - Tth111I (2473) FspI (2457) MscI (2437) PluTI (2358) SfoI (2356) NarI (2355) KasI (2354) BspDI * - ClaI * (2195) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) BglII (609) PaeR7I - XhoI (613) Eco53kI (618) SacI (620) HindIII (622) EcoRI (629) PstI (638) SalI (639) AccI (640) Acc65I (645) KpnI (649) SacII (652) PspOMI (653) ApaI (657) BamHI (660) AgeI (666) PaqCI (670) BsmBI - Esp3I (696) BsgI (698) BbsI (958) NotI (999) MfeI (1105) HpaI (1118) BtsI - BtsαI (1194) AflII (1237) DraIII (1471) CsiI - SexAI * (1944) SfiI (2130) StuI (2176) pPTunerC 4331 bp
AflIII  (4273)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4273)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3453)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BsaI  (3344)
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  (3130)
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  (2871)
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  (2588)
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  (2473)
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  (2473)
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.
FspI  (2457)
1 site
T G C G C A A C G C G T
MscI  (2437)
1 site
T G G C C A A C C G G T
PluTI  (2358)
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.
SfoI  (2356)
1 site
G G C G C C C C G C G G
NarI  (2355)
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.
KasI  (2354)
1 site
G G C G C C C C G C G G
BspDI  (2195)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2195)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SnaBI  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
BglII  (609)
1 site
A G A T C T T C T A G A
PaeR7I  (613)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (613)
1 site
C T C G A G G A G C T C
Eco53kI  (618)
1 site
G A G C T C C T C G A G
SacI  (620)
1 site
G A G C T C C T C G A G
HindIII  (622)
1 site
A A G C T T T T C G A A
EcoRI  (629)
1 site
G A A T T C C T T A A G
PstI  (638)
1 site
C T G C A G G A C G T C
SalI  (639)
1 site
G T C G A C C A G C T G
AccI  (640)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
Acc65I  (645)
1 site
G G T A C C C C A T G G
KpnI  (649)
1 site
G G T A C C C C A T G G
SacII  (652)
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.
PspOMI  (653)
1 site
G G G C C C C C C G G G
ApaI  (657)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BamHI  (660)
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.
AgeI  (666)
1 site
A C C G G T T G G C C A
PaqCI  (670)
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.
BsmBI  (696)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (696)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BsgI  (698)
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).
BbsI  (958)
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.
NotI  (999)
1 site
G C G G C C G C C G C C G G C G
MfeI  (1105)
1 site
C A A T T G G T T A A C
HpaI  (1118)
1 site
G T T A A C C A A T T G
BtsI  (1194)
1 site
G C A G T G N N C G T C A C
BtsαI  (1194)
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  (1237)
1 site
C T T A A G G A A T T C
DraIII  (1471)
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  (1944)
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  (1944)
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.
SfiI  (2130)
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.
StuI  (2176)
1 site
A G G C C T T C C G G A
NeoR/KanR
2227 .. 3021  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2227 .. 3021  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
3629 .. 4217  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3629 .. 4217  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1247 .. 1702  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1247 .. 1702  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1835 .. 2192  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1835 .. 2192  =  358 bp
SV40 enhancer and early promoter
DD-C
674 .. 997  =  324 bp
107 amino acids  =  11.6 kDa
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
FKBP12 mutant optimized for use as a C-terminal tag
DD-C
674 .. 997  =  324 bp
107 amino acids  =  11.6 kDa
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
FKBP12 mutant optimized for use as a C-terminal tag
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
1119 .. 1240  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1119 .. 1240  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1729 .. 1833  =  105 bp
AmpR promoter
1729 .. 1833  =  105 bp
MCS
591 .. 671  =  81 bp
multiple cloning site of fluorescent protein plasmids
MCS
591 .. 671  =  81 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3253 .. 3300  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3253 .. 3300  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2043 .. 2178  =  136 bp
SV40 origin of replication
SV40 ori
2043 .. 2178  =  136 bp
SV40 origin of replication
ORF:  2227 .. 3021  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  758 .. 997  =  240 bp
ORF:  79 amino acids  =  8.6 kDa
ORF:  2399 .. 2785  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3042 .. 3491  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2536 .. 3072  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3247 .. 3480  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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