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"
ORIGIN
        1 acgcgttcga gctcgccccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
       61 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      181 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      241 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      301 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      421 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      481 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
      541 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      601 atccagcctc cgggggatct tggtggcgtg aaactcccgc acctcttcgg ccagcgaatt
      661 cagggtcctg tggacagctc acctagctgc aatggctaca ggctcccgga cgtccctgct
      721 cctggctttt ggcctgctct gcctgccctg gcttcaagag ggcagtgcct ctagaggagt
      781 gcaggtggag actatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg
      841 cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa atggattcct cccgggacag
      901 aaacaagccc tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg
      961 ggttgcccag atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta
     1021 tggtgccact gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga
     1081 gcttctaaaa ctggaaacta gaggagtgca ggtggagact atctccccag gagacgggcg
     1141 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     1201 aaagaaaatg gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     1261 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     1321 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     1381 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaactagag gagtgcaggt
     1441 ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt
     1501 gcactacacc gggatgcttg aagatggaaa gaaaatggat tcctcccggg acagaaacaa
     1561 gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc
     1621 ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc
     1681 cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct
     1741 aaaactggaa actagaggag tgcaggtgga aaccatctcc ccgggagacg ggcgcacctt
     1801 ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa
     1861 aatggattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt
     1921 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac
     1981 tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc
     2041 cactctcgtc ttcgatgtgg agcttctaaa actggaaact agtgctagaa accgtcagaa
     2101 gagattccca accattccct taagcagact ttttgacaac gctatgctcc gcgcccatcg
     2161 tctgcaccag ctggcctttg acacctacca ggagtttgaa gaagcctata tcccaaagga
     2221 acagaagtat tcattcctgc agaaccccca gacctccctc tgtttctcag agtctattcc
     2281 gacaccctcc aacagggagg aaacacaaca gaaatccaac ctagagctgc tccgcatctc
     2341 cctgctgctc atccagtcgt ggctggagcc cgtgcagttc ctcaggagtg tcttcgccaa
     2401 cagcctggtg tacggcgcct ctgacagcaa cgtctatgac ctcctaaagg acctagagga
     2461 aggcatccaa acgctgatgg ggaggctgga agatggcagc ccccggactg ggcagatctt
     2521 caagcagacc tacagcaagt tcgacacaaa ctcacacaac gatgacgcac tactcaagaa
     2581 ctacgggctg ctctactgct tcaggaagga catggacaag gtcgagacat tcctgcgcat
     2641 cgtgcagtgc cgctctgtgg agggcagctg tggcttctag cccgggatcc tgagaacttc
     2701 agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta
     2761 tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac
     2821 catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct
     2881 cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa
     2941 cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt
     3001 ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt
     3061 gttataatta aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat
     3121 attcttattg gtagaaacaa ctacatcctg gtcatcatcc tgcctttctc tttatggtta
     3181 caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct
     3241 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttgtt
     3301 gtgctgtctc atcattttgg caaaggattc actcctcagg tgcaggctgc ctatcagaag
     3361 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     3421 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     3481 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     3541 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     3601 tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc agtatatgaa
     3661 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3721 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     3781 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     3841 cttatgaaga tccctcgagg agctttttgc aaaagcccta ggcctccaaa aaagcctctt
     3901 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     3961 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     4021 ggagttaggg gcgggactat ggttgctgac taattgctgc attaatgaat cggccaacgc
     4081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5701 ttaaaagtgc tcatcattgc cacctccacc tgaaaacgtt cttcggggcg aaaactctca
     5761 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5821 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5881 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5941 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6001 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6061 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6121 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6181 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6241 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6301 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6361 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     6421 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     6481 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     6541 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     6601 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     6661 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     6721 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     6781 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc
     6841 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6901 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6961 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gg
//
