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Plasmid Files

pLVX-Hom-Nuc1

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

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pLVX-Hom-Nuc1 Sequence and MappLVX-Hom-Nuc1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 XcmI (8356) AanI (8169) SgrDI (8131) SspI (8014) PvuI (7580) FspI (7432) AhdI (7210) DrdI (6428) BmtI (5936) NheI (5932) AanI (5885) Bsu36I (5142) PstI (5026) MluI (4408) SexAI * (4375) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) PpuMI - SanDI (1934) cPPT/CTS BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) EcoRI (2803) PaeR7I - XhoI (2809) 5' MCS SpeI (2815) SV40 NLS XbaI (2848) NotI (3177) 3' MCS BamHI (3184) SnaBI (3192) PspOMI (3315) ApaI (3319) AvrII (3353) BmgBI (3745) PflFI - Tth111I (3850) BsiWI (3864) RsrII (3924) BstEII (3942) MscI * (4183) pLVX-Hom-Nuc1 8499 bp
XcmI  (8356)
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  (8169)
2 sites
T T A T A A A A T A T T
SgrDI  (8131)
1 site
C G T C G A C G G C A G C T G C
SspI  (8014)
1 site
A A T A T T T T A T A A
PvuI  (7580)
1 site
C G A T C G G C T A G C
FspI  (7432)
1 site
T G C G C A A C G C G T
AhdI  (7210)
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  (6428)
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  (5936)
1 site
G C T A G C C G A T C G
NheI  (5932)
1 site
G C T A G C C G A T C G
AanI  (5885)
2 sites
T T A T A A A A T A T T
Bsu36I  (5142)
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  (5026)
1 site
C T G C A G G A C G T C
MluI  (4408)
1 site
A C G C G T T G C G C A
SexAI  (4375)
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
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
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.
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  (2803)
1 site
G A A T T C C T T A A G
PaeR7I  (2809)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2809)
1 site
C T C G A G G A G C T C
SpeI  (2815)
1 site
A C T A G T T G A T C A
XbaI  (2848)
1 site
T C T A G A A G A T C T
NotI  (3177)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3184)
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  (3192)
2 sites
T A C G T A A T G C A T
PspOMI  (3315)
1 site
G G G C C C C C C G G G
ApaI  (3319)
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  (3353)
1 site
C C T A G G G G A T C C
BmgBI  (3745)
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  (3850)
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  (3850)
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.
BsiWI  (3864)
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  (3924)
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  (3942)
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  (4183)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
AmpR
7137 .. 7997  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7137 .. 7928  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7137 .. 7997  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7929 .. 7997  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7137 .. 7997  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5217 .. 5850  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5217 .. 5850  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
3808 .. 4407  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3808 .. 4407  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
4421 .. 5009  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4421 .. 5009  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6381 .. 6966  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6381 .. 6966  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
3202 .. 3775  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
3202 .. 3775  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
DmrB
2854 .. 3174  =  321 bp
107 amino acids  =  11.8 kDa
Product: F36V mutant of FK506-binding protein
FKBP12
binds synthetic ligands that do not bind wild-type
FKBP
DmrB
2854 .. 3174  =  321 bp
107 amino acids  =  11.8 kDa
Product: F36V mutant of FK506-binding protein
FKBP12
binds synthetic ligands that do not bind wild-type
FKBP
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
8150 .. 8284  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8150 .. 8284  =  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
7998 .. 8102  =  105 bp
AmpR promoter
7998 .. 8102  =  105 bp
SV40 NLS
2824 .. 2844  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
2824 .. 2844  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
5' MCS
2803 .. 2820  =  18 bp
multiple cloning site upstream of DmrB
5' MCS
2803 .. 2820  =  18 bp
multiple cloning site upstream of DmrB
3' MCS
3176 .. 3189  =  14 bp
multiple cloning site downstream of DmrB
3' MCS
3176 .. 3189  =  14 bp
multiple cloning site downstream of DmrB
ATG
2821 .. 2823  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2821 .. 2823  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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