LOCUS       V003591                 8474 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003591
VERSION     V003591
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8474)
  AUTHORS   Belmont BJ, Niles JC.
  TITLE     Inducible Control of Subcellular RNA Localization Using a Synthetic
            Protein-RNA Aptamer Interaction
  JOURNAL   PLoS ONE 7 (10), E46868 (2012)
   PUBMED   23056498
REFERENCE   2  (bases 1 to 8474)
  AUTHORS   Belmont BJ, Niles JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2012) Biological Engineering, Massachusetts
            Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139,
            USA
REFERENCE   3  (bases 1 to 8474)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8474)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
            date: "2012"; volume: "7"; issue: "10"; pages: "E46868"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (12-SEP-2012) Biological Engineering, Massachusetts Institute of
            Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..416
                     /label="URA3 promoter"
     CDS             417..1217
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1951..1967
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        1977..1995
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2269..2386
                     /label="UAS"
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
     CDS             2735..4009
                     /gene="SHE3"
                     /label="SWI5-dependent HO expression protein 3"
                     /note="SWI5-dependent HO expression protein 3 from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P38272"
     CDS             4028..4645
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label="TetR"
                     /note="TetR binds to the tetracycline operator tetO to
                     inhibit transcription. This inhibition can be relieved by
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     CDS             complement(4650..4676)
                     /label="9xHis"
                     /note="9xHis affinity tag"
     CDS             4676..5389
                     /label="EGFP"
                     /note="enhanced GFP"
     promoter        complement(5700..5718)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5739..5755)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5763..5779)
                     /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)."
     promoter        complement(5787..5817)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5832..5853)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6141..6729)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6903..7760)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7761..7865)
                     /label="AmpR promoter"
     misc_feature    7902..8405
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgacatcgat ggagcaattc
     2041 ttactttttt tttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac
     2101 catatacata tccatataca tatccatata catatccata tacatatcca tatctaatct
     2161 tacttatatg ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa aaccttctct
     2221 ttggaacttt cagtaatacg cttaactgct cattgctata ttgaagtacg gattagaagc
     2281 cgccgagcgg gtgacagccc tccgaaggaa gactctcctc cgtgcgtcct cgtcttcacc
     2341 ggtcgcgttc ctgaaacgca gatgtgcctc gcgccgcact gctccgaaca ataaagattc
     2401 tacaatacta gcttttatgg ttatgaagag gaaaaattgg cagtaacctg gccccacaaa
     2461 ccttcaaatg aacgaatcaa attaacaacc ataggatgat aatgcgatta gttttttagc
     2521 cttatttctg gggtaattaa tcagcgaagc gatgattttt gatcccctta catttacata
     2581 tatataaact tgcataaatt ggtcaatgca agaaatacat atttggtctt ttctaattcg
     2641 tagtttttca agttcttaga tgctttcttt ttctcttttt tacagatatc aaggatccaa
     2701 ttatctactt aagaacacaa aactcgacta caacatgtcg gaccaggata atacccagac
     2761 ttcttcaagc aagttggcac ctcaccataa tatttttatg gcaaacttgg aaagttcccc
     2821 cacaaaagat agaaatacct ccagccaaaa tgcctcatct tcgagagtca ttgaatcact
     2881 gcatgaccag attgatatgc tgacaaaaac aaatctacaa ttgactacgc aatctcaaaa
     2941 tttattgagt aaattggaat tagcacaatc caaagaatca aaactactgg aaaatttaaa
     3001 tctgctgaag aacgagaacg aaaatctcaa ctcgatattt gaaagaaaga acaagaagct
     3061 aaaagagcta gagaaagatt atagcgaatt gagtaaccgt tacaatgaac aaaaggagaa
     3121 aatggatcaa ttgagtaaat tggcgaaaaa ttctagtgcc attgaacaat cttgctctga
     3181 aaaattgcaa aatatggaag ttaactataa ttcattattg gagtcacaaa atctttacag
     3241 agaccactat tctgatgaaa tttccaaatt aaatgaaaaa atcggtttgt tggagttaga
     3301 gttaagtaat caaaatttaa attatggttc agatactagt tcaaattcag atatagaatt
     3361 gaatttaaat aaattcaatg actctgtgaa agatttgaaa tctctggaaa cagaaaagga
     3421 ttcaaaatta agcaagataa tcactcactc tttagatgag ttaaatctac aaagctggct
     3481 aaatctgtac cagacgaacg aaaatctgat atcaactttt gctgaaaaga tggatttgaa
     3541 agacgtcttg aaaagaaacg atgagaagat tagtaataaa ggcgcggtag tacaaacctt
     3601 gaagaagaat gtacaaaccc aggtggaaag caacaacgct gacgctttga gtagcaataa
     3661 tgcacaggat atgcttccta ttaagatggt caaattaaga aagacgccga acacaaacga
     3721 ttcatcctcc aatggtaata gcagtaacaa taaaagaaga agtttctata ctgcgtcgcc
     3781 tttgctgtca tcgggttcta ttccaaaatc tgcatccccg gttttgcctg gtgttaaaag
     3841 aactgcctca gtaagaaaac caagctcgag cagtagtaaa acaaatgtaa cgcataataa
     3901 cgatccaagt acatctccca caatctcagt gcctcccggt gttacaagaa ctgtttcctc
     3961 cactcataag aaaaagggaa atagtatggt tgttcacggg gcccaatcca gcggtggtgg
     4021 tggtggtatg tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     4081 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca
     4141 gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga
     4201 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     4261 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     4321 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     4381 ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg
     4441 gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     4501 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     4561 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     4621 aaaacaactt aaatgtgaaa gtgggtccgg tggtggtggt ggtggtggtg gtggtgtgag
     4681 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4741 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4801 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4861 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4921 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4981 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5041 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5101 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5161 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5221 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5281 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5341 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc tcgagggtgg
     5401 gcgcgcctaa attccttcga tagatcaatt tttttctttt ctctttcccc atcctttacg
     5461 ctaaaataat agtttatttt attttttgaa tattttttat ttatatacgt atatatagac
     5521 tattatttat cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt
     5581 aatgccttta tgcagttttt ttttcccatt cgatatttct atgttcgggt tcagcgtatt
     5641 ttaagtttaa taactcgaaa attctgcgtt cgttaaagct tgagctccag cttttgttcc
     5701 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     5761 aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc
     5821 tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5881 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5941 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6001 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6061 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6121 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6181 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6241 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6301 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6361 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6481 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6541 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6601 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6661 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6721 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6781 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6841 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6901 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6961 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7021 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7081 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7141 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7201 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7261 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7321 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7381 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7441 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7501 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7561 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7621 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7681 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7741 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7801 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7861 ccgcgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata
     7921 attataattt aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa
     7981 attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa
     8041 caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat
     8101 cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta
     8161 atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg
     8221 ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc
     8281 taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga
     8341 gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag
     8401 cgatccgtcc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8461 gaggcccttt cgtc
//
