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pLVX-Het-1

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

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pLVX-Het-1 Sequence and MappLVX-Het-1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 XcmI (8294) AanI (8107) SgrDI (8069) SspI (7952) PvuI (7518) FspI (7370) AhdI (7148) DrdI (6366) BmtI (5874) NheI (5870) AanI (5823) Bsu36I (5080) PstI (4964) MluI (4346) NruI * (833) FseI (1150) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) SanDI (1934) cPPT/CTS BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) EcoRI (2804) PaeR7I - XhoI (2810) 5' MCS SpeI (2816) ATG XbaI (2828) SphI (2939) NotI (3115) 3' MCS BamHI (3122) SnaBI (3130) AvrII (3291) AarI - BfuAI - BspMI (3479) BmgBI (3683) PflFI - Tth111I (3788) TspMI - XmaI (3795) SmaI (3797) BsiWI (3802) RsrII (3862) BstEII (3880) MscI * (4121) SexAI * (4313) pLVX-Het-1 8437 bp
XcmI  (8294)
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.
AanI  (8107)
2 sites
T T A T A A A A T A T T
SgrDI  (8069)
1 site
C G T C G A C G G C A G C T G C
SspI  (7952)
1 site
A A T A T T T T A T A A
PvuI  (7518)
1 site
C G A T C G G C T A G C
FspI  (7370)
1 site
T G C G C A A C G C G T
AhdI  (7148)
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  (6366)
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  (5874)
1 site
G C T A G C C G A T C G
NheI  (5870)
1 site
G C T A G C C G A T C G
AanI  (5823)
2 sites
T T A T A A A A T A T T
Bsu36I  (5080)
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  (4964)
1 site
C T G C A G G A C G T C
MluI  (4346)
1 site
A C G C G T T G C G C A
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

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
SanDI  (1934)
1 site
G G G W C C C C C C W G G G

Sticky ends from different SanDI 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)
2 sites
T A C G T A A T G C A T
EcoRI  (2804)
1 site
G A A T T C C T T A A G
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  (2828)
1 site
T C T A G A A G A T C T
SphI  (2939)
1 site
G C A T G C C G T A C G
NotI  (3115)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3122)
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.
SnaBI  (3130)
2 sites
T A C G T A A T G C A T
AvrII  (3291)
1 site
C C T A G G G G A T C C
AarI  (3479)
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.
BfuAI  (3479)
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  (3479)
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.
BmgBI  (3683)
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  (3788)
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  (3788)
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.
TspMI  (3795)
1 site
C C C G G G G G G C C C
XmaI  (3795)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (3797)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BsiWI  (3802)
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  (3862)
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  (3880)
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  (4121)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
SexAI  (4313)
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.
AmpR
7075 .. 7935  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7075 .. 7866  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7075 .. 7935  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7867 .. 7935  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7075 .. 7935  =  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
5155 .. 5788  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5155 .. 5788  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
3746 .. 4345  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3746 .. 4345  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
4359 .. 4947  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4359 .. 4947  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6319 .. 6904  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6319 .. 6904  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
3140 .. 3713  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
3140 .. 3713  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
FRB* (DmrC)
2834 .. 3112  =  279 bp
93 amino acids  =  11.3 kDa
Product: T2098L mutant of FKBP-rapamycin binding
domain of human FRAP
binds synthetic ligands such as AP21967
FRB* (DmrC)
2834 .. 3112  =  279 bp
93 amino acids  =  11.3 kDa
Product: T2098L mutant of FKBP-rapamycin binding
domain of human FRAP
binds synthetic ligands such as AP21967
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
8088 .. 8222  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8088 .. 8222  =  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
7936 .. 8040  =  105 bp
AmpR promoter
7936 .. 8040  =  105 bp
5' MCS
2804 .. 2821  =  18 bp
multiple cloning site upstream of DmrC
5' MCS
2804 .. 2821  =  18 bp
multiple cloning site upstream of DmrC
3' MCS
3114 .. 3127  =  14 bp
multiple cloning site downstream of DmrC
3' MCS
3114 .. 3127  =  14 bp
multiple cloning site downstream of DmrC
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
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