Basic Vector Information
prHom-Nuc1 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
prHom-Nuc1 vector Sequence
LOCUS prHom-Nuc1. 6064 bp DNA circular SYN 01-JAN-1980
DEFINITION Mammalian vector encoding three FKBP homodimerizer domains, for
creating nuclear oligomers that can be dissociated with a drug. Also
known as pC4EN-FM3.
ACCESSION .
VERSION .
KEYWORDS prHom-Nuc1.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6064)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6064)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Clone into the XbaI site to target the partner protein to the
nucleus and to fuse three copies of DmrD to its C-terminus.
FEATURES Location/Qualifiers
source 1..6064
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 19..322
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 323..526
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
5'UTR 616..655
/label=HSV TK 5'-UTR
/note="5' untranslated region from the herpes simplex virus
thymidine kinase gene"
CDS 673..675
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 685..711
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 733..753
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 760..1080
/label=DmrD
/note="dimeric F36M mutant of FK506-binding protein FKBP12"
CDS 1087..1407
/codon_start=1
/product="dimeric F36M mutant of FK506-binding protein
FKBP12"
/label=dimeric F36M mutant of FK506-binding protein
FK
/note="DmrD"
/note="dimerization can be reversed by ligand binding"
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
CDS 1414..1734
/codon_start=1
/product="dimeric F36M mutant of FK506-binding protein
FKBP12"
/label=dimeric F36M mutant of FK506-binding protein
FK
/note="DmrD"
/note="dimerization can be reversed by ligand binding"
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
intron 1766..2338
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
polyA_signal 2535..2590
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(2943..3078)
/direction=LEFT
/label=SV40 ori
/note="SV40 origin of replication"
rep_origin complement(3355..3943)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4114..4986)
/codon_start=1
/product="beta-lactamase"
/label=beta-lactamase
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFQVEVAMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDL
VEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAE
IGASLIKHW"
promoter complement(4987..5091)
/label=AmpR promoter
rep_origin complement(5423..5878)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6023..6039
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 6046..6064
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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