pLVX-PTuner Green

Lentiviral vector for fusing a destabilization domain to the N-terminus of a partner protein, with co-expression of ZsGreen1.

Sequence Author: Clontech (TaKaRa)

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SgrDI (8169) SspI (8052) PvuI (7618) FspI (7470) DrdI (6463) BmtI (5972) NheI (5968) StuI (5957) Bsu36I (5178) PstI (5063) PfoI (4731) MluI (4445) BsrGI (4299) BssHII (711) NruI * (833) FseI (1150) MfeI (1189) BbvCI - Bpu10I (1424) BstAPI (1453) AleI (1577) KflI - PpuMI (1934) cPPT/CTS BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) SwaI (2797) TspMI - XmaI (2923) SmaI (2925) AgeI (3129) EcoRI (3135) PaeR7I - XhoI (3141) SpeI (3147) MCS XbaI (3153) NotI (3160) BamHI (3167) PspOMI (3288) ApaI (3292) AvrII (3326) BclI * (3831) SgrAI (3836) PflFI - Tth111I (3983) pLVX-PTuner Green 8536 bp
SgrDI  (8169)
1 site
C G T C G A C G G C A G C T G C
SspI  (8052)
1 site
A A T A T T T T A T A A
PvuI  (7618)
1 site
C G A T C G G C T A G C
FspI  (7470)
1 site
T G C G C A A C G C G T
DrdI  (6463)
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  (5972)
1 site
G C T A G C C G A T C G
NheI  (5968)
1 site
G C T A G C C G A T C G
StuI  (5957)
1 site
A G G C C T T C C G G A
Bsu36I  (5178)
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  (5063)
1 site
C T G C A G G A C G T C
PfoI  (4731)
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.
MluI  (4445)
1 site
A C G C G T T G C G C A
BsrGI  (4299)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BssHII  (711)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
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.
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

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
BstAPI  (1453)
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.
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.
TspMI  (2923)
1 site
C C C G G G G G G C C C
XmaI  (2923)
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  (2925)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
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
BclI  (3831)
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.
SgrAI  (3836)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PflFI  (3983)
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  (3983)
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.
AmpR
7175 .. 8035  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7175 .. 7966  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7175 .. 8035  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7967 .. 8035  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7175 .. 8035  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ZsGreen1
3749 .. 4444  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
3749 .. 4444  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
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
5253 .. 5886  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5253 .. 5886  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
4458 .. 5046  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4458 .. 5046  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6416 .. 7004  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6416 .. 7004  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
3175 .. 3747  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
3175 .. 3747  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
DD
2805 .. 3128  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   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
2 segments
   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
8188 .. 8322  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8188 .. 8322  =  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
8036 .. 8140  =  105 bp
AmpR promoter
8036 .. 8140  =  105 bp
MCS
3129 .. 3172  =  44 bp
multiple cloning site
MCS
3129 .. 3172  =  44 bp
multiple cloning site
ORF:  4561 .. 4857  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  3749 .. 4444  =  696 bp
ORF:  231 amino acids  =  26.1 kDa
ORF:  2805 .. 3287  =  483 bp
ORF:  160 amino acids  =  17.5 kDa
ORF:  4548 .. 5075  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  7305 .. 7571  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7175 .. 8035  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  4126 .. 4419  =  294 bp
ORF:  97 amino acids  =  10.9 kDa
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