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pLVX-PTunerC

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

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pLVX-PTunerC.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SgrDI (8065) SspI (7948) PvuI (7514) FspI (7366) AhdI (7144) DrdI (6359) AflIII - PciI (6251) Bsu36I (5073) PflMI (4594) SexAI * (4307) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI (1424) AleI (1577) KflI (1934) BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) AfeI (2798) PaeR7I - XhoI (2815) BstBI (2829) EcoRI (2831) PspOMI (2855) ApaI (2859) BamHI (2862) AarI (2876) SphI (3259) AgeI (3330) SpeI (3465) BlpI (3493) PasI (3789) BsiWI (3796) RsrII (3856) BstEII (3874) MscI * (4115) pLVX-PTunerC 8385 bp
SgrDI  (8065)
1 site
C G T C G A C G G C A G C T G C
SspI  (7948)
1 site
A A T A T T T T A T A A
PvuI  (7514)
1 site
C G A T C G G C T A G C
FspI  (7366)
1 site
T G C G C A A C G C G T
AhdI  (7144)
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  (6359)
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.
AflIII  (6251)
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  (6251)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
Bsu36I  (5073)
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.
PflMI  (4594)
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.
SexAI  (4307)
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
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.
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
AfeI  (2798)
1 site
A G C G C T T C G C G A
PaeR7I  (2815)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2815)
1 site
C T C G A G G A G C T C
BstBI  (2829)
1 site
T T C G A A A A G C T T
EcoRI  (2831)
1 site
G A A T T C C T T A A G
PspOMI  (2855)
1 site
G G G C C C C C C G G G
ApaI  (2859)
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  (2862)
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.
AarI  (2876)
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 AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
SphI  (3259)
1 site
G C A T G C C G T A C G
AgeI  (3330)
1 site
A C C G G T T G G C C A
SpeI  (3465)
1 site
A C T A G T T G A T C A
BlpI  (3493)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
PasI  (3789)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
BsiWI  (3796)
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  (3856)
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  (3874)
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  (4115)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
AmpR
7071 .. 7931  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7071 .. 7862  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7071 .. 7931  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7863 .. 7931  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7071 .. 7931  =  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
5148 .. 5781  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5148 .. 5781  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
3740 .. 4339  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3740 .. 4339  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
4353 .. 4941  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4353 .. 4941  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6312 .. 6900  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6312 .. 6900  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
3220 .. 3719  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
3220 .. 3719  =  500 bp
mouse phosphoglycerate kinase 1 promoter
DD-C
2880 .. 3203  =  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
2880 .. 3203  =  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
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
8084 .. 8218  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8084 .. 8218  =  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
7932 .. 8036  =  105 bp
AmpR promoter
7932 .. 8036  =  105 bp
MCS
2796 .. 2867  =  72 bp
multiple cloning site
MCS
2796 .. 2867  =  72 bp
multiple cloning site
ORF:  4456 .. 4752  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  7201 .. 7467  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  3500 .. 4339  =  840 bp
ORF:  279 amino acids  =  30.1 kDa
ORF:  2964 .. 3203  =  240 bp
ORF:  79 amino acids  =  8.6 kDa
ORF:  4443 .. 4970  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  7071 .. 7931  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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