pLVX-Het-2

Lentiviral vector encoding an FKBP heterodimerizer domain, for creating soluble fusion proteins that can be dimerized with a drug.

Sequence Author: Clontech (TaKaRa)

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XcmI (8759) SgrDI (8534) SspI (8417) FspI (7835) AhdI (7613) BmtI (6339) NheI (6335) StuI (6324) Bsu36I (5545) PfoI (5097) MluI (4811) RsrII (4193) KasI (637) NarI (638) SfoI (639) PluTI (641) BssHII (711) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) KflI - PpuMI (1934) cPPT/CTS BspDI - ClaI (2180) PaeR7I - XhoI (2810) 5' MCS SpeI (2816) ATG XbaI (2825) TspMI - XmaI (2946) SmaI (2948) NotI (3154) 3' MCS BamHI (3161) PspOMI (3292) ApaI (3296) AvrII (3330) PaqCI (3518) BmgBI (3722) PshAI (3812) AsiSI (4149) pLVX-Het-2 8902 bp
XcmI  (8759)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SgrDI  (8534)
1 site
C G T C G A C G G C A G C T G C
SspI  (8417)
1 site
A A T A T T T T A T A A
FspI  (7835)
1 site
T G C G C A A C G C G T
AhdI  (7613)
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.
BmtI  (6339)
1 site
G C T A G C C G A T C G
NheI  (6335)
1 site
G C T A G C C G A T C G
StuI  (6324)
1 site
A G G C C T T C C G G A
Bsu36I  (5545)
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.
PfoI  (5097)
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  (4811)
1 site
A C G C G T T G C G C A
RsrII  (4193)
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.
KasI  (637)
1 site
G G C G C C C C G C G G
NarI  (638)
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.
SfoI  (639)
1 site
G G C G C C C C G C G G
PluTI  (641)
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.
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.
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

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.
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
PaeR7I  (2810)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2810)
1 site
C T C G A G G A G C T C
SpeI  (2816)
1 site
A C T A G T T G A T C A
XbaI  (2825)
1 site
T C T A G A A G A T C T
TspMI  (2946)
1 site
C C C G G G G G G C C C
XmaI  (2946)
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  (2948)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NotI  (3154)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3161)
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  (3292)
1 site
G G G C C C C C C G G G
ApaI  (3296)
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  (3330)
1 site
C C T A G G G G A T C C
PaqCI  (3518)
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.
BmgBI  (3722)
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.
PshAI  (3812)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
AsiSI  (4149)
1 site
G C G A T C G C C G C T A G C G
HygR
3791 .. 4810  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
3791 .. 4810  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
AmpR
7540 .. 8400  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7540 .. 8331  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7540 .. 8400  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8332 .. 8400  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7540 .. 8400  =  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
5620 .. 6253  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5620 .. 6253  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
4824 .. 5412  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4824 .. 5412  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6784 .. 7369  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6784 .. 7369  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
3179 .. 3752  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
3179 .. 3752  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
ATG
2822 .. 2824  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2822 .. 2824  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
FKBP (DmrA)
2831 .. 3151  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
2831 .. 3151  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein 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
8553 .. 8687  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8553 .. 8687  =  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
8401 .. 8505  =  105 bp
AmpR promoter
8401 .. 8505  =  105 bp
3' MCS
3153 .. 3166  =  14 bp
multiple cloning site downstream of DmrA
3' MCS
3153 .. 3166  =  14 bp
multiple cloning site downstream of DmrA
5' MCS
2810 .. 2821  =  12 bp
multiple cloning site upstream of DmrA
5' MCS
2810 .. 2821  =  12 bp
multiple cloning site upstream of DmrA
ORF:  4927 .. 5223  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  2822 .. 3175  =  354 bp
ORF:  117 amino acids  =  12.9 kDa
ORF:  3662 .. 4810  =  1149 bp
ORF:  382 amino acids  =  42.6 kDa
ORF:  7670 .. 7936  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4914 .. 5441  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  3677 .. 4543  =  867 bp
ORF:  288 amino acids  =  32.7 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  2587 .. 2850  =  264 bp
ORF:  87 amino acids  =  9.6 kDa
ORF:  7540 .. 8400  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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