LOCUS       40924_49472        9485 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector YCplac22 YCp-She3-TG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9485)
  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 9485)
  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 9485)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9485)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2012"; volume: "7"; issue: "10"; pages: "E46868"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-SEP-2012) Biological Engineering, Massachusetts Institute of 
            Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1239..2513
                     /gene="SHE3"
                     /label=SHE3
                     /note="SWI5-dependent HO expression protein 3 from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c). 
                     Accession#: P38272"
     CDS             2532..3149
                     /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(3154..3180)
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             3180..3893
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             4320..4340
                     /label=7xHis
                     /note="6xHis affinity tag"
     primer_bind     complement(4921..4937)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(5696..6858)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     promoter        complement(7372..7473)
                     /label=TRP1 promoter
     promoter        7579..7683
                     /label=AmpR promoter
     CDS             7684..8541
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8715..9303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttta
      241 agtttttgtt cccatagact ccctattagc cgctttggtg cgctttggta ggtggtaaac
      301 caaaatgtcc tttagtccta gataactctt tttatctaaa agaatattct gcaagattat
      361 agcgtcccat gactcttcga cattattttt ttctccagga ttataagagg aatgttgaaa
      421 atttgccctt ttataaaaac caggccacat tacagtaaat gatctattcg ttacagtttc
      481 tagattcaaa taaatatcat tccagaacgg aataacgaat tcgttcaaca aatccgtgag
      541 tttgcggaag tccttgttaa atccgctgaa tgacccattg agtagcttga agaacctgta
      601 actaagccat aagtaagcac tgagatactg taaaggtatg gcactctgat cgtcgctgat
      661 attcgaaatt gctagactga tgaaaaatct tgattccacc ttaatacaaa aaataatatc
      721 attgttcagc tcaataattt tctctaaatc ttcgccgtcc ttgtttacgc atgaactcgt
      781 aagtgaccaa atgccctgta ttactcctat gatggacagt ttttcgttta aagagggtat
      841 tgaatcagta gtgccaaaat aatggaacag cagcaactgc ttgaaagtgt cattttcgtt
      901 ggtagatctt gatggatcaa aaaccgtaat atagtgtagt ctcatatccg tctttcatat
      961 atcaatttaa tagttgtacg atttagaatg tatttttggt tccacgtcaa atgaaactat
     1021 ttccctattg ctactacata ttattgtcat cgcaataaaa taggctaaat agttacccgc
     1081 tcgtttcgtc ttttgttcgg gcgagagctt cattactaac tactattgca ttttgtttat
     1141 tgatttgttg ccatgagtag cagcagtctg aaggggttac caaacgttag ctcgtctatc
     1201 aagcacgcca aggttcaacg acactacttt tgtgtaagat gtcggaccag gataataccc
     1261 agacttcttc aagcaagttg gcacctcacc ataatatttt tatggcaaac ttggaaagtt
     1321 cccccacaaa agatagaaat acctccagcc aaaatgcctc atcttcgaga gtcattgaat
     1381 cactgcatga ccagattgat atgctgacaa aaacaaatct acaattgact acgcaatctc
     1441 aaaatttatt gagtaaattg gaattagcac aatccaaaga atcaaaacta ctggaaaatt
     1501 taaatctgct gaagaacgag aacgaaaatc tcaactcgat atttgaaaga aagaacaaga
     1561 agctaaaaga gctagagaaa gattatagcg aattgagtaa ccgttacaat gaacaaaagg
     1621 agaaaatgga tcaattgagt aaattggcga aaaattctag tgccattgaa caatcttgct
     1681 ctgaaaaatt gcaaaatatg gaagttaact ataattcatt attggagtca caaaatcttt
     1741 acagagacca ctattctgat gaaatttcca aattaaatga aaaaatcggt ttgttggagt
     1801 tagagttaag taatcaaaat ttaaattatg gttcagatac tagttcaaat tcagatatag
     1861 aattgaattt aaataaattc aatgactctg tgaaagattt gaaatctctg gaaacagaaa
     1921 aggattcaaa attaagcaag ataatcactc actctttaga tgagttaaat ctacaaagct
     1981 ggctaaatct gtaccagacg aacgaaaatc tgatatcaac ttttgctgaa aagatggatt
     2041 tgaaagacgt cttgaaaaga aacgatgaga agattagtaa taaaggcgcg gtagtacaaa
     2101 ccttgaagaa gaatgtacaa acccaggtgg aaagcaacaa cgctgacgct ttgagtagca
     2161 ataatgcaca ggatatgctt cctattaaga tggtcaaatt aagaaagacg ccgaacacaa
     2221 acgattcatc ctccaatggt aatagcagta acaataaaag aagaagtttc tatactgcgt
     2281 cgcctttgct gtcatcgggt tctattccaa aatctgcatc cccggttttg cctggtgtta
     2341 aaagaactgc ctcagtaaga aaaccaagct cgagcagtag taaaacaaat gtaacgcata
     2401 ataacgatcc aagtacatct cccacaatct cagtgcctcc cggtgttaca agaactgttt
     2461 cctccactca taagaaaaag ggaaatagta tggttgttca cggggcccaa tccagcggtg
     2521 gtggtggtgg tatgtctaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     2581 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     2641 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     2701 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     2761 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     2821 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     2881 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
     2941 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     3001 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     3061 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     3121 tagaaaaaca acttaaatgt gaaagtgggt ccggtggtgg tggtggtggt ggtggtggtg
     3181 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3241 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3301 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3361 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     3421 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3481 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3541 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3601 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3661 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3721 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3781 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3841 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagctcgagg
     3901 gtgggcgcgc ctaaaatatg ccgacacaag ggagtatata tataggattc atttagataa
     3961 tagaacaaaa gtaaatagat gcctctaagg ttttatagac acgaatccat atttttcact
     4021 ttcggtcact cacaacggtt ttttctctac tataatgtgg ctgaaattaa acgaacaggc
     4081 caaaatgtgg tacgcaatag tagccagtag gtgaattaac cataaataat tacgctaatt
     4141 ttgccttggt aaatttcata agaccaatga tagcaggtgc gactgctccg tcaagtcagc
     4201 atgaaatttt gatcgcatcg aatctcatca agaagccatc aacgtctcaa aataaaacgc
     4261 cgacagctca atccagttca ggtaacaatg gagctgctga tggtgcaccg cagggttacc
     4321 atcatcacca tcatcaccat cgtcatttat ggtggcctcg gacaacagac catcaatatt
     4381 ggtgtgtctt gaggaaaaac caatttgcct attacaaaac tcgggatgaa agagaggcta
     4441 taagtgtcat accgagattt gacatactta attttaagat aagcgagctc gatggtatat
     4501 taactgttta tactccatcc aaggatttaa tattcaaatt tccacgaggg caaaatgaga
     4561 aggttggtat ggaactgatg cataattgga aaattgccct tgaaaaattc ctctctagtc
     4621 ctagtggcaa cgaaagtgtc actacaggta gtgattatga tgaagaagag gatgatgatg
     4681 atttaattgt agttgacgaa aaggcgggtc cttctagcag caagcattct tgcagtttga
     4741 cgatggatga gcaactgtct cgtgaagata aagaatttta taggatgttc gatccaagaa
     4801 atgcagagca ccaggtttgt tctgggattc tttacacaaa agtgaaaaag aagaaactgt
     4861 tcaatagggc taaatggcaa aagttcaatg tggaattgac taatacttcg ttcagaattc
     4921 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4981 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5041 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     5101 tacgcatctg tgcggtattt cacaccgcat atatcgctgg gccattctca tgaagaatat
     5161 cttgaattta ttgtcatatt actagttggt gtggaagtcc atatatcggt gatcaatata
     5221 gtggttgaca tgctggctag tcaacattga gccttttgat catgcaaata tattacggta
     5281 ttttacaatc aaatatcaaa cttaactatt gactttataa cttatttagg tggtaacatt
     5341 cttataaaaa agaaaaaaat tactgcaaaa cagtactagc ttttaacttg tatcctaggt
     5401 tatctatgct gtctcaccat agagaatatt acctatttca gaatgtatgt ccatgattcg
     5461 ccgggtaaat acatataata cacaaatctg gcttaataaa gtctataata tatctcataa
     5521 agaagtgcta aattggctag tgctatatat ttttaagaaa atttcttttg actaagtcca
     5581 tatcgacttt gtaaaagttc actttagcat acatatatta cacgagccag aaattgtaac
     5641 ttttgcctaa aatcacaaat tgcaaaattt aattgcttgc aaaaggtcac atgcttataa
     5701 tcaacttttt taaaaattta aaatactttt ttatttttta tttttaaaca taaatgaaat
     5761 aatttattta ttgtttatga ttaccgaaac ataaaacctg ctcaagaaaa agaaactgtt
     5821 ttgtccttgg aaaaaaagca ctacctagga gcggccaaaa tgccgaggct ttcatagctt
     5881 aaactcttta cagaaaatag gcattataga tcagttcgag ttttcttatt cttccttccg
     5941 gttttatcgt cacagtttta cagtaaataa gtatcacctc ttagagttcg atgataagct
     6001 gtcaaacatg agaattaatt ccacatgtta aaatagtgaa ggagcatgtt cggcacacag
     6061 tggaccgaac gtggggtaag tgcactaggg tccggttaaa cggatctcgc attgatgagg
     6121 caacgctaat tatcaacata tagattgtta tctatctgca tgaacacgaa atctttactt
     6181 gacgacttga ggctgatggt gtttatgcaa agaaaccact gtgtttaata tgtgtcactg
     6241 tttgatatta ctgtcagcgt agaagataat agtaaaagcg gttaataagt gtatttgaga
     6301 taagtgtgat aaagttttta cagcgaaaag acgataaata caagaaaatg attacgagga
     6361 tacggagaga ggtatgtaca tgtgtattta tatactaagc tgccggcggt tgtttgcaag
     6421 accgagaaaa ggctagcaag aatcgggtca ttgtagcgta tgcgcctgtg aacattctct
     6481 tcaacaagtt tgattccatt gcggtgaaat ggtaaaagtc aaccccctgc gatgtatatt
     6541 ttcctgtaca atcaatcaaa aagccaaatg atttagcatt atctttacat cttgttattt
     6601 tacagatttt atgtttagat cttttatgct tgcttttcaa aaggcttgca ggcaagtgca
     6661 caaacaatac ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa
     6721 atttgctatt ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac
     6781 accaataacg ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc
     6841 taacataaaa tgtaagctct cggggctctc ttgccttcca acccagtcag aaatcgagtt
     6901 ccaatccaaa agttcacctg tcccacctgc ttctgaatca aacaagggaa taaacgaatg
     6961 aggtttctgt gaagctgcac tgagtagtat gttgcagtct tttggaaata cgagtctttt
     7021 aataactggc aaaccgagga actcttggta ttcttgccac gactcatctc catgcagttg
     7081 gacgatatca atgccgtaat cattgaccag agccaaaaca tcctccttag gttgattacg
     7141 aaacacgcca accaagtatt tcggagtgcc tgaactattt ttatatgctt ttacaagact
     7201 tgaaattttc cttgcaataa ccgggtcaat tgttctcttt ctattgggca cacatataat
     7261 acccagcaag tcagcatcgg aatctagtgc acattctgcg gcctctgtgc tctgcaagcc
     7321 gcaaactttc accaatggac cagaactacc tgtgaaatta ataacagaca tactccaagc
     7381 tgcctttgtg tgcttaatca cgtatactca cgtgctcaat agtcaccaat gccctccctc
     7441 ttggccctct ccttttcttt tttcgaccga attaattctt gaagacgaaa gggcctcgtg
     7501 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     7561 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7621 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7681 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7741 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7801 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7861 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7921 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7981 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     8041 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     8101 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     8161 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     8221 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     8281 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8341 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8401 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8461 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8521 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8821 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     8881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     9001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     9061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     9121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     9181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     9241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     9301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     9361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     9421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     9481 gaaga
//
