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pLVX-Het-Mem1

Lentiviral vector encoding FKBP heterodimerizer domains, for creating membrane-associated fusion proteins that can be dimerized with a drug. 

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pLVX-Het-Mem1 Sequence and MappLVX-Het-Mem1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 XcmI (9125) AanI (8938) SgrDI (8900) SspI (8783) FspI (8201) AhdI (7979) BmtI (6705) NheI (6701) StuI (6690) AanI (6654) PfoI (5463) MluI (5177) RsrII (4559) BssHII (711) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) SanDI (1934) BspDI - ClaI (2180) CMV enhancer SnaBI (2480) PaeR7I - XhoI (2810) 5' MCS SpeI (2816) N-myristoylation signal AfeI (2861) XbaI (2864) TspMI - XmaI (2985) SmaI (2987) SphI (3283) NotI (3520) BamHI (3527) SnaBI (3535) PspOMI (3658) ApaI (3662) AvrII (3696) PshAI (4178) AsiSI (4515) pLVX-Het-Mem1 9268 bp
XcmI  (9125)
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  (8938)
2 sites
T T A T A A A A T A T T
SgrDI  (8900)
1 site
C G T C G A C G G C A G C T G C
SspI  (8783)
1 site
A A T A T T T T A T A A
FspI  (8201)
1 site
T G C G C A A C G C G T
AhdI  (7979)
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  (6705)
1 site
G C T A G C C G A T C G
NheI  (6701)
1 site
G C T A G C C G A T C G
StuI  (6690)
1 site
A G G C C T T C C G G A
AanI  (6654)
2 sites
T T A T A A A A T A T T
PfoI  (5463)
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  (5177)
1 site
A C G C G T T G C G C A
RsrII  (4559)
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.
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

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.
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
SnaBI  (2480)
2 sites
T A C G T A A T G C A T
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
AfeI  (2861)
1 site
A G C G C T T C G C G A
XbaI  (2864)
1 site
T C T A G A A G A T C T
TspMI  (2985)
1 site
C C C G G G G G G C C C
XmaI  (2985)
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  (2987)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SphI  (3283)
1 site
G C A T G C C G T A C G
NotI  (3520)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3527)
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  (3535)
2 sites
T A C G T A A T G C A T
PspOMI  (3658)
1 site
G G G C C C C C C G G G
ApaI  (3662)
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  (3696)
1 site
C C T A G G G G A T C C
PshAI  (4178)
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  (4515)
1 site
G C G A T C G C C G C T A G C G
HygR
4157 .. 5176  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
4157 .. 5176  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
7906 .. 8766  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7906 .. 8697  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7906 .. 8766  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8698 .. 8766  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7906 .. 8766  =  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
5986 .. 6619  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5986 .. 6619  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
5190 .. 5778  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5190 .. 5778  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7150 .. 7735  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7150 .. 7735  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
3545 .. 4118  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
3545 .. 4118  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
FKBP (DmrA)
2870 .. 3190  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
2870 .. 3190  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP (DmrA)
3197 .. 3517  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
synthetic gene with alternative codons
FKBP (DmrA)
3197 .. 3517  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
synthetic gene with alternative codons
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
8919 .. 9053  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8919 .. 9053  =  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
8767 .. 8871  =  105 bp
AmpR promoter
8767 .. 8871  =  105 bp
N-myristoylation signal
2822 .. 2863  =  42 bp
14 amino acids  =  1.5 kDa
Product: N-myristoylation signal from Src kinase
N-myristoylation signal
2822 .. 2863  =  42 bp
14 amino acids  =  1.5 kDa
Product: N-myristoylation signal from Src kinase
3' MCS
3519 .. 3532  =  14 bp
multiple cloning site downstream of DmrA
3' MCS
3519 .. 3532  =  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
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