LOCUS       V008480                 8379 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008480
VERSION     V008480
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8379)
  AUTHORS   Kraft C.
  TITLE     A toolbox for the analysis of weak and low abundant molecule
            interactions in vivo by M-Track in budding yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8379)
  AUTHORS   Brezovich A, Schuschnig M, Ammerer G, Kraft C.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2014) Department of Biochemistry and Cell Biology,
            MFPL, University of Vienna, Dr. Bohr Gasse 9, Vienna 1030, Austria
REFERENCE   3  (bases 1 to 8379)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8379)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-AUG-2014) Department of Biochemistry and Cell Biology, MFPL,
            University of Vienna, Dr. Bohr Gasse 9, Vienna 1030, Austria"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    155..176
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        191..221
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    229..245
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     253..269
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        290..308
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      345..993
                     /gene="ATG18"
                     /regulatory_class="promoter"
     CDS             994..2493
                     /gene="ATG18"
                     /label="Autophagy-related protein 18"
                     /note="Autophagy-related protein 18 from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P43601"
     CDS             2554..2574
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             2605..2778
                     /label="ProtA"
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
     CDS             2782..2952
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label="ProtA"
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNALIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     misc_feature    3019..3312
                     /gene="ATG18"
                     /label="4x Histone3"
                     /note="4x Histone3"
     CDS             3337..3426
                     /label="3xHA"
                     /note="three tandem HA epitope tags"
     CDS             3445..3462
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             3466..3537
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags"
     CDS             3538..3561
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      3584..3831
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     promoter        complement(3850..3868)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3878..3894)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      4035..4490
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4753..5409)
                     /label="HIS3"
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     promoter        complement(5410..5596)
                     /label="HIS3 promoter"
     misc_feature    complement(5982..6485)
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        6522..6626
                     /label="AmpR promoter"
     CDS             6627..7484
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7658..8246
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
       61 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      121 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      181 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      241 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      301 actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctaacgt agttatccga
      361 atgcataaat tagactagta tgtgacaacg ctaaacatgc gttccccaga accacctgcg
      421 gttgttgaga ggaataggtt ttgcgcttgt ctatggacac tttcttgaac tcaatccaac
      481 tgagcaatac ttaacaatat tgcgattact tgttcttaca taaagagtaa gaaatacttg
      541 cgaaataaac ggtaatcaat caaatttagg gtcataagtt gtattcgcac aatatgcata
      601 agcactgact caagaaagaa gatggttttc gaatggggag ctagcaaaaa gagtcacccc
      661 taagtataaa aactgaatta caaagctctt cgactacttt ctgcaagtaa atcattcgga
      721 agtgcgacaa tataatgaca aaagacgata agttacttct attaattgaa ctttaagtat
      781 ttggctttta tttttttttt tcattttctt actcgcgctt ttctttatta agggctttct
      841 caattagatg ttaataatgt cataaggcat cgttattccg ctttatattg gtaactcagc
      901 agtggtcatt gccttcgttc acgaccctcc cttattaatc agttagtaat agtgttccag
      961 ttaactctgt atccttttct cttcggcctg acaatgtctg attcatcacc tactatcaac
     1021 tttattaatt tcaatcaaac cggaacgtgt atttcccttg gaacgtcgaa aggtttcaaa
     1081 atattcaatt gtgagccctt cggaaaattt tattcagagg acagtggggg ctatgctatc
     1141 gtcgagatgt tgttctccac ctcgttacta gccctcgttg ggataggcga tcaacctgcg
     1201 ctttcaccaa ggagattgcg tataatcaac acaaaaaaac attctattat ctgtgaggtg
     1261 actttcccta cttctattct gagtgtgaaa atgaataagt ctcgattggt ggtactttta
     1321 caagagcaga tttatattta tgatatcaac accatgagac tattgcatac tatagaaaca
     1381 aaccctaacc cacgtggcct tatggctatg tctccttcgg tagccaacag ctatttagtg
     1441 tatccatcac caccaaaagt tattaactcc gaaattaaag ctcatgccac cacaaacaat
     1501 atcacattgt cagttggtgg caacacagag accagtttca agagagatca gcaagatgct
     1561 ggccatagtg acattagcga cttggatcag tattcgagct ttactaagag ggatgatgcg
     1621 gatccaacaa gcagtaacgg cggtaacagc agtataataa agaatggtga tgtgatcgta
     1681 ttcaacttgg aaacattaca gccaaccatg gtcatcgaag ctcataaggg cgagattgct
     1741 gcaatggcaa ttagttttga tgggacacta atggctaccg cctctgataa aggtactatc
     1801 atcagggtct ttgacattga aacgggtgat aagatctacc aattcaggag agggacgtac
     1861 gcgacaagaa tttactccat atcattcagt gaagatagcc agtacttggc ggttaccggc
     1921 tcttccaaaa ccgtgcatat cttcaaattg gggcattcaa tgagcaacaa taaactagac
     1981 agcgatgata gcaacatgga agaagctgca gccgatgatt catcgctcga taccaccagt
     2041 atcgatgcgc tgagtgacga ggaaaacccg acaagactcg caagagaacc atatgtggat
     2101 gcatcaagaa agacaatggg taggatgata cgttactctt ctcaaaagct atcccgaaga
     2161 gctgccagaa cattgggtca gattttcccc atcaaagtta catcgttgtt ggaatcctcg
     2221 cgccattttg cgtctttgaa acttcccgtt gaaaccaatt cccatgtaat gaccatatca
     2281 agtataggct ctccaataga tatagacaca tccgagtatc cggaactctt cgaaactggc
     2341 aattccgcaa gtacagagtc ctaccatgag cctgttatga agatggtccc catcagggtc
     2401 gtttcctcgg atggatacct atacaacttt gttatggacc cggagagagg cggcgattgc
     2461 ttaatattgt cacagtattc catcttgatg gacctgcagg agaaaatctc atcctccggg
     2521 gcacttgatt atgatattcc aactactgct agcgagaatt tgtattttca gggagaattc
     2581 ggccttgcgc aacacgatga agccgtggac aacaaattca acaaagaaca acaaaacgcg
     2641 ttctatgaga tcttacattt acctaactta aacgaagaac aacgaaacgc cttcatccaa
     2701 agtttaaaag atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat
     2761 gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag aacaacaaaa cgcgttctat
     2821 gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccctcat ccaaagttta
     2881 aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatgatgct
     2941 caggcgccga aagtagacgc gaatcatcag ctgcaggacg acctcgagtc taggggatcc
     3001 ccggggatcc cgaattcggc tcgtactaag cagaccgctc gcaagtccac cggcggcaag
     3061 gccccgcgca agcagctggc ccagatcccg aattcggctc gtactaagca gaccgctcgc
     3121 aagtccaccg gcggcaaggc cccgcgcaag cagctggccc agatcccgaa ttcggctcgt
     3181 actaagcaga ccgctcgcaa gtccaccggc ggcaaggccc cgcgcaagca gctggcccag
     3241 atcccgaatt cggctcgtac taagcagacc gctcgcaagt ccaccggcgg caaggccccg
     3301 cgcaagcagc tggcccagat ctgcggacgc atcttttacc catacgatgt tcctgactat
     3361 gcgggctatc cctatgacgt cccggactat gcaggatcct atccatatga cgttccagat
     3421 tacgctgctc agtgcgtcga gtcccaccac catcatcacc acggagatta taaagacgat
     3481 gacgataaag attataagga tgatgatgac aaagattaca aagatgacga tgataaggac
     3541 tacaaagacg acgatgataa gctgcagtaa cgtcgacctc gagtcatgta attagttatg
     3601 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     3661 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     3721 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     3781 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac
     3841 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     3901 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3961 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4021 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4081 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4141 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4201 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4261 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4321 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4381 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4441 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg
     4501 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagatccgtc gagttcaaga
     4561 gaaaaaaaaa gaaaaagcaa aaagaaaaaa ggaaagcgcg cctcgttcag aatgacacgt
     4621 atagaatgat gcattacctt gtcatcttca gtatcatact gttcgtatac atacttactg
     4681 acattcatag gtatacatat atacacatgt atatatatcg tatgctgcag ctttaaataa
     4741 tcggtgtcac tacataagaa cacctttggt ggagggaaca tcgttggtac cattgggcga
     4801 ggtggcttct cttatggcaa ccgcaagagc cttgaacgca ctctcactac ggtgatgatc
     4861 attcttgcct cgcagacaat caacgtggag ggtaattctg ctagcctctg caaagctttc
     4921 aagaaaatgc gggatcatct cgcaagagag atctcctact ttctcccttt gcaaaccaag
     4981 ttcgacaact gcgtacggcc tgttcgaaag atctaccacc gctctggaaa gtgcctcatc
     5041 caaaggcgca aatcctgatc caaacctttt tactccacgc gccagtaggg cctctttaaa
     5101 agcttgaccg agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc
     5161 accaatgcac tcaacgatta gcgaccagcc ggaatgcttg gccagagcat gtatcatatg
     5221 gtccagaaac cctatacctg tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc
     5281 tactgcttct gcctcttttt ctgggaagat cgagtgctct atcgctaggg gaccaccctt
     5341 taaagagatc gcaatctgaa tcttggtttc atttgtaata cgctttacta gggctttctg
     5401 ctctgtcatc tttgccttcg tttatcttgc ctgctcattt tttagtatat tcttcgaaga
     5461 aatcacatta ctttatataa tgtataattc attatgtgat aatgccaatc gctaagaaaa
     5521 aaaaagagtc atccgctagg ggaaaaaaaa aaatgaaaat cattaccgag gcataaaaaa
     5581 atatagagtg tactagagga ggccaagagt aatagaaaaa gaaaattgcg ggaaaggact
     5641 gtgttatgac ttccctgact aatgccgtgt tcaaacgata cctggcagtg actcctagcg
     5701 ctcaccaagc tcttaaaacg ggaatttatg gtgcactctc agtacaatct gctctgatgc
     5761 cgcatagtta agccagcccc gacacccgcc aacacgcgct gacgcgccct gacgggcttg
     5821 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5881 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5941 tttataggtt aatgtcatga taataatggt ttcttaggac ggatcgcttg cctgtaactt
     6001 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta
     6061 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa
     6121 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat
     6181 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa
     6241 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg
     6301 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct
     6361 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt
     6421 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac
     6481 gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6541 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6601 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6661 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6721 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6781 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6841 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6901 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6961 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7021 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7081 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7141 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7201 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7261 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7321 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7381 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7441 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7501 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7561 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7801 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8281 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8341 accgtattac cgcctttgag tgagctgata ccgctcgcc
//
