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"
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 ccagaagcgc gtatggcttc tagctatcct tatgacgtgc ctgactatgc cagcctggga
      721 ggaccttcta gtcctaagaa gaagagaaag gtgtctagag gagtgcaggt ggagactatc
      781 tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt gcactacacc
      841 gggatgcttg aagatggaaa gaaaatggat tcctcccggg acagaaacaa gccctttaag
      901 tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt
      961 gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac
     1021 ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct aaaactggaa
     1081 actagaggag tgcaggtgga gactatctcc ccaggagacg ggcgcacctt ccccaagcgc
     1141 ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa aatggattcc
     1201 tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc
     1261 tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca
     1321 gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc
     1381 ttcgatgtgg agcttctaaa actggaaact agaggagtgc aggtggagac tatctcccca
     1441 ggagacgggc gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg
     1501 cttgaagatg gaaagaaaat ggattcctcc cgggacagaa acaagccctt taagtttatg
     1561 ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt
     1621 cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc
     1681 atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaactagt
     1741 tattgaggat cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt
     1801 cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat
     1861 gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct
     1921 actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt
     1981 aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg
     2041 taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc
     2101 agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat
     2161 aaattctggc tggcgtggaa atattcttat tggtagaaac aactacatcc tggtcatcat
     2221 cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc
     2281 tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     2341 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaaggat tcactcctca
     2401 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     2461 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     2521 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2581 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     2641 tttggtttag agtttggcaa catatgccat atgctggctg ccatgaacaa aggtggctat
     2701 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     2761 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     2821 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     2881 cagtcatagc tgtccctctt ctcttatgaa gatccctcga ggagcttttt gcaaaagccc
     2941 taggcctcca aaaaagcctc ttcactactt ctggaatagc tcagaggccg aggcggcctc
     3001 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     3061 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgct
     3121 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3181 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3241 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3301 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3361 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3421 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3481 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3541 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3601 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3661 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3721 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3781 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3841 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3901 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3961 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4021 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4081 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4141 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4201 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4261 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4321 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4381 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4441 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4501 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4561 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4621 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4681 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4741 taccgcgcca catagcagaa ctttaaaagt gctcatcatt gccacctcca cctgaaaacg
     4801 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4861 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4921 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4981 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5041 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5101 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5161 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5221 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5281 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5341 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     5401 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     5461 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5521 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5581 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5641 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5701 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5761 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5821 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     5881 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5941 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6001 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     6061 tagg
//
