Basic Vector Information
prHom-Sec1 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-Sec1 vector Sequence
LOCUS prHom-Sec1. 7002 bp DNA circular SYN 01-JAN-1980
DEFINITION Mammalian vector encoding four FKBP homodimerizer domains, for
creating ER-localized oligomers that can be dissociated with a drug.
Also known as pC4S1-FM4-FCS-hGH.
ACCESSION .
VERSION .
KEYWORDS prHom-Sec1.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7002)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7002)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Clone between the SpeI and BamHI sites to replace the stuffer
sequence with the coding sequence of interest. The insert should
encode the furin cleavage site and a stop codon.
FEATURES Location/Qualifiers
source 1..7002
/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"
sig_peptide 692..769
/label=hGH signal sequence
/note="human growth hormone signal sequence"
CDS 776..1096
/label=DmrD
/note="dimeric F36M mutant of FK506-binding protein FKBP12"
CDS 1103..1423
/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 1430..1750
/codon_start=1
/product="dimeric F36M mutant of FK506-binding protein
FKBP12"
/note="DmrD"
/note="dimerization can be reversed by ligand binding"
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
CDS 1757..2077
/codon_start=1
/product="dimeric F36M mutant of FK506-binding protein
FKBP12"
/note="DmrD"
/note="dimerization can be reversed by ligand binding"
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
CDS 2087..2104
/codon_start=1
/product="cleavage site for the furin protease in the
trans-Golgi network"
/label=cleavage site for the furin protease in the
tra
/note="furin cleavage site"
/translation="RNRQKR"
CDS 2105..2677
/codon_start=1
/label=Stuffer sequence
/note="stuffer sequence"
/note="to be replaced by the coding sequence of interest"
/translation="FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFL
QNPQTSLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYG
ASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLL
YCFRKDMDKVETFLRIVQCRSVEGSCGF"
intron 2704..3276
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
polyA_signal 3473..3528
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(3881..4016)
/direction=LEFT
/label=SV40 ori
/note="SV40 origin of replication"
rep_origin complement(4293..4881)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5052..5924)
/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(5925..6029)
/label=AmpR promoter
rep_origin complement(6361..6816)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6961..6977
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 6984..7002
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
This page is informational only.