LOCUS       V003590                 7939 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003590
VERSION     V003590
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7939)
  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 7939)
  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 7939)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7939)
  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..7939
                     /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..3352
                     /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(3357..3383)
                     /label="9xHis"
                     /note="9xHis affinity tag"
     CDS             3383..4096
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             4124..4861
                     /gene="SHE2"
                     /label="SWI5-dependent HO expression protein 2"
                     /note="SWI5-dependent HO expression protein 2 from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P36068"
     promoter        complement(5165..5183)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5204..5220)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5228..5244)
                     /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(5252..5282)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5297..5318)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5606..6194)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6368..7225)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7226..7330)
                     /label="AmpR promoter"
     misc_feature    7367..7870
                     /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 caacatgtct agattagata aaagtaaagt
     2761 gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa
     2821 actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg
     2881 ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc acttttgccc
     2941 tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc
     3001 tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca
     3061 gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa
     3121 tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca
     3181 agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt
     3241 attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg
     3301 ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtccggtgg
     3361 tggtggtggt ggtggtggtg gtgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3421 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3481 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3541 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3601 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3661 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3721 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3781 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3841 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3901 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3961 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4021 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4081 ggacgagctg tacaagctcg aggctggtgc tgctgctggt gctatgagca aagacaaaga
     4141 tatcaaagtc actcctggaa cttgtgaatt agttgaacaa atactcgcct tactctcccg
     4201 gtatctctca tcttatattc acgtgctgaa caagttcatc agtcatttgc gaagggtagc
     4261 tacgctacga ttcgagagaa ctactttgat taaatttgtt aagaaattga gattttacaa
     4321 cgattgtgtg ttaagctata atgcatcaga gtttatcaat gaaggtaaaa acgaattaga
     4381 cccagaggcg gattcgtttg acaaggtaat tttaccaata gcttcaatgt ttgttaaatg
     4441 tgttgaaacg tttgacttac tgaactatta tctaacgcag tcattacaaa aggaaatttt
     4501 atctaaaact ttgaacgagg acctaacgct aacagcagag tccatattag ccattgacga
     4561 tacctacaat catttcgtga aattttctca atggatgatt gagtctttgc gtataggcag
     4621 caacctgcta gatttagaag ttgttcaatt tgccattaaa tgcgcagatg aggacggaac
     4681 gaatatcgga gaaactgata atatcttctt gcaggagatt cttcctgtca actccgaaga
     4741 agaatttcag accttgtcag ctgcgtggca ctccatttta gatggtaagc tgagcgcctt
     4801 ggacgaagaa tttgatgttg tcgctactaa atggcatgac aaatttggta aattgaaaaa
     4861 ctgaggcgcg cctaaattcc ttcgatagat caattttttt cttttctctt tccccatcct
     4921 ttacgctaaa ataatagttt attttatttt ttgaatattt tttatttata tacgtatata
     4981 tagactatta tttatctttt aatgattatt aagattttta ttaaaaaaaa attcgctcct
     5041 cttttaatgc ctttatgcag tttttttttc ccattcgata tttctatgtt cgggttcagc
     5101 gtattttaag tttaataact cgaaaattct gcgttcgtta aagcttgagc tccagctttt
     5161 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     5221 tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc ataaagtgta
     5281 aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc tcactgcccg
     5341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7201 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7261 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7321 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc cttttcatca cgtgctataa
     7381 aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa
     7441 aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc
     7501 gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt
     7561 ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat
     7621 cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa
     7681 gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac
     7741 tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac
     7801 acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag
     7861 gcaagcgatc cgtcctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     7921 atcacgaggc cctttcgtc
//
