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Plasmid Files

pLVX-rHom-Nuc1

Lentiviral vector encoding three FKBP homodimerizer domains, for creating nuclear oligomers that can be dissociated with a drug. 

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pLVX-rHom-Nuc1 Sequence and MappLVX-rHom-Nuc1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 XcmI (9017) AanI (8830) SgrDI (8792) SspI (8675) PvuI (8241) FspI (8093) AhdI (7871) DrdI (7089) BmtI (6597) NheI (6593) AanI (6546) Bsu36I (5803) PstI (5687) MluI (5069) SexAI * (5036) MscI * (4844) BstEII (4603) RsrII (4585) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) PpuMI - SanDI (1934) BspDI - ClaI (2180) CMV enhancer NdeI (2374) SnaBI (2480) EcoRI (2804) 5' MCS PaeR7I - XhoI (2810) ATG SV40 NLS XbaI (2846) NotI (3838) BamHI (3845) SnaBI (3853) PspOMI (3976) ApaI (3980) AvrII (4014) BmgBI (4406) PflFI - Tth111I (4511) BsiWI (4525) pLVX-rHom-Nuc1 9160 bp
XcmI  (9017)
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  (8830)
2 sites
T T A T A A A A T A T T
SgrDI  (8792)
1 site
C G T C G A C G G C A G C T G C
SspI  (8675)
1 site
A A T A T T T T A T A A
PvuI  (8241)
1 site
C G A T C G G C T A G C
FspI  (8093)
1 site
T G C G C A A C G C G T
AhdI  (7871)
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  (7089)
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  (6597)
1 site
G C T A G C C G A T C G
NheI  (6593)
1 site
G C T A G C C G A T C G
AanI  (6546)
2 sites
T T A T A A A A T A T T
Bsu36I  (5803)
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  (5687)
1 site
C T G C A G G A C G T C
MluI  (5069)
1 site
A C G C G T T G C G C A
SexAI  (5036)
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.
MscI  (4844)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BstEII  (4603)
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.
RsrII  (4585)
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.
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  (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
XbaI  (2846)
1 site
T C T A G A A G A T C T
NotI  (3838)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3845)
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  (3853)
2 sites
T A C G T A A T G C A T
PspOMI  (3976)
1 site
G G G C C C C C C G G G
ApaI  (3980)
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  (4014)
1 site
C C T A G G G G A T C C
BmgBI  (4406)
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  (4511)
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  (4511)
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  (4525)
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.
AmpR
7798 .. 8658  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7798 .. 8589  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7798 .. 8658  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8590 .. 8658  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7798 .. 8658  =  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
5878 .. 6511  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5878 .. 6511  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
4469 .. 5068  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4469 .. 5068  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
5082 .. 5670  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5082 .. 5670  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7042 .. 7627  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7042 .. 7627  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
3863 .. 4436  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
3863 .. 4436  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
DmrD
2852 .. 3172  =  321 bp
107 amino acids  =  11.8 kDa
Product: dimeric F36M mutant of FK506-binding
protein FKBP12
dimerization can be reversed by ligand binding
DmrD
2852 .. 3172  =  321 bp
107 amino acids  =  11.8 kDa
Product: dimeric F36M mutant of FK506-binding
protein FKBP12
dimerization can be reversed by ligand binding
DmrD
3179 .. 3499  =  321 bp
107 amino acids  =  11.8 kDa
Product: dimeric F36M mutant of FK506-binding
protein FKBP12
dimerization can be reversed by ligand binding
DmrD
3179 .. 3499  =  321 bp
107 amino acids  =  11.8 kDa
Product: dimeric F36M mutant of FK506-binding
protein FKBP12
dimerization can be reversed by ligand binding
DmrD
3506 .. 3826  =  321 bp
107 amino acids  =  11.8 kDa
Product: dimeric F36M mutant of FK506-binding
protein FKBP12
dimerization can be reversed by ligand binding
DmrD
3506 .. 3826  =  321 bp
107 amino acids  =  11.8 kDa
Product: dimeric F36M mutant of FK506-binding
protein FKBP12
dimerization can be reversed by ligand binding
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
8811 .. 8945  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8811 .. 8945  =  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
8659 .. 8763  =  105 bp
AmpR promoter
8659 .. 8763  =  105 bp
SV40 NLS
2825 .. 2845  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
2825 .. 2845  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
3' MCS
3837 .. 3850  =  14 bp
multiple cloning site downstream of DmrD
3' MCS
3837 .. 3850  =  14 bp
multiple cloning site downstream of DmrD
5' MCS
2804 .. 2815  =  12 bp
multiple cloning site upstream of DmrD
5' MCS
2804 .. 2815  =  12 bp
multiple cloning site upstream of DmrD
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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