pDD

Vector encoding the 12 kDa destabilization domain (DD), a mutant of FKBP12.

Sequence Author: Clontech (TaKaRa)

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SacII (3026) BtgI (3023) KpnI (3022) Acc65I (3018) BanII - SacI (3016) Eco53kI (3014) ApoI - EcoRI (3006) PluTI (2849) SfoI (2847) NarI (2846) KasI (2845) BstAPI (2795) NdeI (2794) PfoI (2656) EcoO109I (2599) AatII (2545) ZraI (2543) SspI (2427) XmnI (2222) ScaI (2103) TsoI (2022) NmeAIII (1771) BsrFI (1703) BsaI (1684) AhdI (1623) BfuAI - BspMI - PaqCI (0) BsgI (28) AvaI - BsoBI - TspMI - XmaI (119) SmaI (121) BbsI (288) PflMI (337) BamHI (341) XbaI (347) PstI (363) SphI (369) HindIII (371) BspQI - SapI (614) AflIII - PciI (730) AlwNI (1146) pDD 3040 bp
SacII  (3026)
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.
BtgI  (3023)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
KpnI  (3022)
1 site
G G T A C C C C A T G G
Acc65I  (3018)
1 site
G G T A C C C C A T G G
BanII  (3016)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (3016)
1 site
G A G C T C C T C G A G
Eco53kI  (3014)
1 site
G A G C T C C T C G A G
ApoI  (3006)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (3006)
1 site
G A A T T C C T T A A G
PluTI  (2849)
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  (2847)
1 site
G G C G C C C C G C G G
NarI  (2846)
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  (2845)
1 site
G G C G C C C C G C G G
BstAPI  (2795)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
NdeI  (2794)
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.
PfoI  (2656)
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.
EcoO109I  (2599)
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.
AatII  (2545)
1 site
G A C G T C C T G C A G
ZraI  (2543)
1 site
G A C G T C C T G C A G
SspI  (2427)
1 site
A A T A T T T T A T A A
XmnI  (2222)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2103)
1 site
A G T A C T T C A T G A
TsoI  (2022)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
NmeAIII  (1771)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrFI  (1703)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
BsaI  (1684)
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  (1623)
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.
BfuAI  (0)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (0)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PaqCI  (0)
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.
BsgI  (28)
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).
AvaI  (119)
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  (119)
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.
TspMI  (119)
1 site
C C C G G G G G G C C C
XmaI  (119)
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  (121)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BbsI  (288)
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.
PflMI  (337)
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.
BamHI  (341)
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  (347)
1 site
T C T A G A A G A T C T
PstI  (363)
1 site
C T G C A G G A C G T C
SphI  (369)
1 site
G C A T G C C G T A C G
HindIII  (371)
1 site
A A G C T T T T C G A A
BspQI  (614)
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  (614)
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.
AflIII  (730)
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  (730)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (1146)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AmpR
1550 .. 2410  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1550 .. 2341  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1550 .. 2410  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2342 .. 2410  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1550 .. 2410  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
791 .. 1379  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
791 .. 1379  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
DD
1 .. 324  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 1:  
   1 .. 3  =  3 bp
   1 amino acid  =  149.2 Da
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTunerâ„¢ system
L106P mutant of FKBP12
DD
1 .. 324  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 2:  
   4 .. 324  =  321 bp
   107 amino acids  =  11.8 kDa
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTunerâ„¢ system
L106P mutant of FKBP12
DD
1 .. 324  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTunerâ„¢ system
L106P mutant of FKBP12
AmpR promoter
2411 .. 2515  =  105 bp
AmpR promoter
2411 .. 2515  =  105 bp
M13 rev
389 .. 405  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
389 .. 405  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2989 .. 3005  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2989 .. 3005  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  3035 .. 351  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  1680 .. 1946  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2796 .. 3038  =  243 bp
ORF:  80 amino acids  =  9.4 kDa
ORF:  1550 .. 2410  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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