prHom-Sec1 vector (V011088)

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"