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Plasmid Files

pLVX-PTuner

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

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pLVX-PTuner.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SgrDI (8105) SspI (7988) PvuI (7554) FspI (7406) AhdI (7184) DrdI (6399) BmtI (5908) NheI (5904) Bsu36I (5114) PstI (4999) MluI (4381) SexAI * (4348) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) KflI - PpuMI (1934) BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) SwaI (2797) AgeI (3129) EcoRI (3135) PaeR7I - XhoI (3141) SpeI (3147) XbaI (3153) NotI (3160) BamHI (3167) PspOMI (3288) ApaI (3292) AvrII (3326) BmgBI (3718) PflFI - Tth111I (3823) BsiWI (3837) RsrII (3897) BstEII (3915) MscI * (4156) pLVX-PTuner 8472 bp
SgrDI  (8105)
1 site
C G T C G A C G G C A G C T G C
SspI  (7988)
1 site
A A T A T T T T A T A A
PvuI  (7554)
1 site
C G A T C G G C T A G C
FspI  (7406)
1 site
T G C G C A A C G C G T
AhdI  (7184)
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.
DrdI  (6399)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BmtI  (5908)
1 site
G C T A G C C G A T C G
NheI  (5904)
1 site
G C T A G C C G A T C G
Bsu36I  (5114)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PstI  (4999)
1 site
C T G C A G G A C G T C
MluI  (4381)
1 site
A C G C G T T G C G C A
SexAI  (4348)
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.
NruI  (833)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
FseI  (1150)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
EcoNI  (1170)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
MfeI  (1189)
1 site
C A A T T G G T T A A C
BbvCI  (1424)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1424)
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.
AleI  (1577)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (1934)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1934)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BspDI  (2180)
1 site
A T C G A T T A G C T A
ClaI  (2180)
1 site
A T C G A T T A G C T A
NdeI  (2374)
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  (2480)
1 site
T A C G T A A T G C A T
SwaI  (2797)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AgeI  (3129)
1 site
A C C G G T T G G C C A
EcoRI  (3135)
1 site
G A A T T C C T T A A G
PaeR7I  (3141)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3141)
1 site
C T C G A G G A G C T C
SpeI  (3147)
1 site
A C T A G T T G A T C A
XbaI  (3153)
1 site
T C T A G A A G A T C T
NotI  (3160)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3167)
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.
PspOMI  (3288)
1 site
G G G C C C C C C G G G
ApaI  (3292)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (3326)
1 site
C C T A G G G G A T C C
BmgBI  (3718)
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.
PflFI  (3823)
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  (3823)
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.
BsiWI  (3837)
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.
RsrII  (3897)
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.
BstEII  (3915)
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.
MscI  (4156)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
AmpR
7111 .. 7971  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7111 .. 7902  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7111 .. 7971  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7903 .. 7971  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7111 .. 7971  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
3' LTR
5189 .. 5822  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5189 .. 5822  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
3781 .. 4380  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3781 .. 4380  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
4394 .. 4982  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4394 .. 4982  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6352 .. 6940  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6352 .. 6940  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
3175 .. 3748  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
3175 .. 3748  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
DD
2805 .. 3128  =  324 bp
108 amino acids  =  11.9 kDa
   Segment 1:  
   2805 .. 2807  =  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
2805 .. 3128  =  324 bp
108 amino acids  =  11.9 kDa
   Segment 2:  
   2808 .. 3128  =  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
2805 .. 3128  =  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
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
8124 .. 8258  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8124 .. 8258  =  135 bp
SV40 polyadenylation signal
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
7972 .. 8076  =  105 bp
AmpR promoter
7972 .. 8076  =  105 bp
MCS
3129 .. 3172  =  44 bp
multiple cloning site
MCS
3129 .. 3172  =  44 bp
multiple cloning site
ORF:  3658 .. 4380  =  723 bp
ORF:  240 amino acids  =  26.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  4484 .. 5011  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  7241 .. 7507  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2805 .. 3287  =  483 bp
ORF:  160 amino acids  =  17.5 kDa
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