pLVX-rHom-Nuc1

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

Sequence Author: Clontech (TaKaRa)

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XcmI (9017) SgrDI (8792) SspI (8675) PvuI (8241) FspI (8093) AhdI (7871) DrdI (7089) BmtI (6597) NheI (6593) Bsu36I (5803) PstI (5687) MluI (5069) CsiI - SexAI * (5036) MscI * (4844) BstEII (4603) RsrII (4585) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) KflI - PpuMI (1934) cPPT/CTS BspDI - ClaI (2180) NdeI (2374) EcoRI (2804) 5' MCS PaeR7I - XhoI (2810) SV40 NLS XbaI (2846) NotI (3838) 3' MCS BamHI (3845) 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.
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
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
CsiI  (5036)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
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

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
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.
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.
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.
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
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
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
AmpR
7798 .. 8658  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   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
2 segments
   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
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
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
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)
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
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
ORF:  5185 .. 5481  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  2822 .. 3859  =  1038 bp
ORF:  345 amino acids  =  38.2 kDa
ORF:  4346 .. 5068  =  723 bp
ORF:  240 amino acids  =  26.2 kDa
ORF:  7928 .. 8194  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5172 .. 5699  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  7798 .. 8658  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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