pPTuner

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

Sequence Author: Clontech (TaKaRa)

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PciI (4264) EcoO109I (3444) BsaI (3335) PfoI (3121) RsrII (2862) BsrDI (2579) PflFI - Tth111I (2464) FspI (2448) MscI (2428) PluTI (2349) SfoI (2347) NarI (2346) KasI (2345) EagI (2252) BspDI * - ClaI * (2186) StuI (2167) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) PaqCI (605) BsmBI - Esp3I (631) BsgI (633) BbsI (893) BglII (930) PaeR7I - XhoI (934) Eco53kI (939) SacI (941) HindIII (943) EcoRI (950) PstI (959) SalI (960) AccI (961) Acc65I (966) KpnI (970) SacII (973) PspOMI (974) ApaI (978) BamHI (981) XbaI * (993) BclI * (1003) MfeI (1096) HpaI (1109) BtsI - BtsαI (1185) MluI (1232) DraIII (1462) CsiI - SexAI * (1935) SfiI (2121) BseRI (2164) pPTuner 4322 bp
PciI  (4264)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3444)
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  (3335)
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  (3121)
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  (2862)
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  (2579)
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  (2464)
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  (2464)
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  (2448)
1 site
T G C G C A A C G C G T
MscI  (2428)
1 site
T G G C C A A C C G G T
PluTI  (2349)
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  (2347)
1 site
G G C G C C C C G C G G
NarI  (2346)
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  (2345)
1 site
G G C G C C C C G C G G
EagI  (2252)
1 site
C G G C C G G C C G G C
BspDI  (2186)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2186)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2167)
1 site
A G G C C T T C C G G A
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
AgeI  (600)
1 site
A C C G G T T G G C C A
PaqCI  (605)
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  (631)
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  (631)
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  (633)
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  (893)
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.
BglII  (930)
1 site
A G A T C T T C T A G A
PaeR7I  (934)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (934)
1 site
C T C G A G G A G C T C
Eco53kI  (939)
1 site
G A G C T C C T C G A G
SacI  (941)
1 site
G A G C T C C T C G A G
HindIII  (943)
1 site
A A G C T T T T C G A A
EcoRI  (950)
1 site
G A A T T C C T T A A G
PstI  (959)
1 site
C T G C A G G A C G T C
SalI  (960)
1 site
G T C G A C C A G C T G
AccI  (961)
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  (966)
1 site
G G T A C C C C A T G G
KpnI  (970)
1 site
G G T A C C C C A T G G
SacII  (973)
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  (974)
1 site
G G G C C C C C C G G G
ApaI  (978)
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  (981)
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.
XbaI  (993)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1003)
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.
MfeI  (1096)
1 site
C A A T T G G T T A A C
HpaI  (1109)
1 site
G T T A A C C A A T T G
BtsI  (1185)
1 site
G C A G T G N N C G T C A C
BtsαI  (1185)
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.
MluI  (1232)
1 site
A C G C G T T G C G C A
DraIII  (1462)
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  (1935)
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  (1935)
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  (2121)
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  (2164)
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.
NeoR/KanR
2218 .. 3012  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2218 .. 3012  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
3620 .. 4208  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3620 .. 4208  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1238 .. 1693  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1238 .. 1693  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1826 .. 2183  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1826 .. 2183  =  358 bp
SV40 enhancer and early promoter
DD
606 .. 929  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 1:  
   606 .. 608  =  3 bp
   1 amino acid  =  149.2 Da
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
606 .. 929  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 2:  
   609 .. 929  =  321 bp
   107 amino acids  =  11.8 kDa
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
606 .. 929  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
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
1110 .. 1231  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1110 .. 1231  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1720 .. 1824  =  105 bp
AmpR promoter
1720 .. 1824  =  105 bp
MCS
930 .. 1008  =  79 bp
multiple cloning site
MCS
930 .. 1008  =  79 bp
multiple cloning site
HSV TK poly(A) signal
3244 .. 3291  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3244 .. 3291  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2034 .. 2169  =  136 bp
SV40 origin of replication
SV40 ori
2034 .. 2169  =  136 bp
SV40 origin of replication
ORF:  2218 .. 3012  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2390 .. 2776  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  606 .. 1001  =  396 bp
ORF:  131 amino acids  =  14.1 kDa
ORF:  3033 .. 3482  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2527 .. 3063  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3238 .. 3471  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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