LOCUS       40924_49477        8721 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector YCplac22 YCp-TG-She2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8721)
  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 8721)
  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 8721)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8721)
  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..8721
                     /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             1021..1638
                     /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(1643..1669)
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             1669..2382
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             2410..3147
                     /gene="SHE2"
                     /label=SHE2
                     /note="SWI5-dependent HO expression protein 2 from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c). 
                     Accession#: P36068"
     CDS             complement(3718..3735)
                     /label=tetracysteine tag
                     /note="tetracysteine peptide that binds biarsenical
                     labeling reagents"
     primer_bind     complement(4157..4173)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(4932..6094)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     promoter        complement(6608..6709)
                     /label=TRP1 promoter
     promoter        6815..6919
                     /label=AmpR promoter
     CDS             6920..7777
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7951..8539
                     /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 ccaagcttgc
      241 atgcctgcag atccgaaatt cgagctgcat acgactttcc aggttccgga tcatcttcgg
      301 agttgccatt aaaaaagggc gatattgtat ttataagcag agatgaacct agtggttggt
      361 ctttagcaaa gcttctcgat ggaagcaaag aaggttgggt tcctactgct tatatgactc
      421 cttataaaga cacaagaaat acggtgcctg ttgcagcgac aggggcagtg aatgacgtta
      481 caaatcagaa atcaagccaa atcgacaata ccatttcaag tgcacaagaa ggtgtacagt
      541 ttggaagtgc aacagttgga cccactagtg ataatcagag taatccggtg gggacgttta
      601 gcgacggttt agcttctgca ttggcagcaa gagcaaacaa aatgcgggcg gaaagtgctg
      661 acgatgatga taatgacgat ggcgatgatg atgatgactg gtaattaaca tgaatatata
      721 gatgtttagt ctcttataga atggttcttc gtgcatgcct cttaacatac caaaagtgca
      781 ttacatatac ttttaaacat actctcgcag aaagttaaag tccctttttt tttttttcat
      841 tttttgtttg tattttcccg ttttgaaaaa ctcccacttt aatgcaggac atcgttcaaa
      901 acaagcttta aagatttgac agtaaaatcc tcactttact caagacattt ttctcaaaaa
      961 aagagaaagc acagtaaacc ctccttaatt ttccttttgc ataataccag acacttaaaa
     1021 atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
     1081 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca
     1141 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
     1201 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
     1261 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
     1321 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
     1381 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt
     1441 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
     1501 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
     1561 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
     1621 cttaaatgtg aaagtgggtc cggtggtggt ggtggtggtg gtggtggtgt gagcaagggc
     1681 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1741 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1801 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1861 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1921 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1981 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2041 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2101 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2161 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2221 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     2281 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2341 accgccgccg ggatcactct cggcatggac gagctgtaca agctcgaggc tggtgctgct
     2401 gctggtgcta tgagcaaaga caaagatatc aaagtcactc ctggaacttg tgaattagtt
     2461 gaacaaatac tcgccttact ctcccggtat ctctcatctt atattcacgt gctgaacaag
     2521 ttcatcagtc atttgcgaag ggtagctacg ctacgattcg agagaactac tttgattaaa
     2581 tttgttaaga aattgagatt ttacaacgat tgtgtgttaa gctataatgc atcagagttt
     2641 atcaatgaag gtaaaaacga attagaccca gaggcggatt cgtttgacaa ggtaatttta
     2701 ccaatagctt caatgtttgt taaatgtgtt gaaacgtttg acttactgaa ctattatcta
     2761 acgcagtcat tacaaaagga aattttatct aaaactttga acgaggacct aacgctaaca
     2821 gcagagtcca tattagccat tgacgatacc tacaatcatt tcgtgaaatt ttctcaatgg
     2881 atgattgagt ctttgcgtat aggcagcaac ctgctagatt tagaagttgt tcaatttgcc
     2941 attaaatgcg cagatgagga cggaacgaat atcggagaaa ctgataatat cttcttgcag
     3001 gagattcttc ctgtcaactc cgaagaagaa tttcagacct tgtcagctgc gtggcactcc
     3061 attttagatg gtaagctgag cgccttggac gaagaatttg atgttgtcgc tactaaatgg
     3121 catgacaaat ttggtaaatt gaaaaactga ggccttcagt ttttagctca aaaagagcaa
     3181 ataagtacca ctagttaata gaacatatat atatacatga atagcctcat cacatagcac
     3241 agaaagtaag ctttcaaaac actgatgatc ggctcttatt ctggcgtact cattttcagc
     3301 atctcaaacc gcgctgtttt tttctcactt tttgatacaa taggtatgca aaggagaatg
     3361 gcaggtgtgc caaggaaaaa aaatatgagg aactatagcc taaaatgaag cctcctctga
     3421 tgtgggaggg accccaattt aggtcaataa tgatttttta aatttatatt cttgtcttaa
     3481 tgaaacttga gtcaagtcga ccagtttcct gacgattttg atatagaata atattgctcg
     3541 tttggccggc aacagattac ataagaagca gatggatgat ataagcggaa ggcaaacttt
     3601 acctcgaata aaccgtttgt tggagcacgt gggaaatccc caagatagtt tgtcaatcct
     3661 acatatagcg ggaacaaatg gtaaggagac agtatctaaa tttttgacat ccatattgca
     3721 gcatccgggg caacagcggc aaagggtctt gattggtagg tatactacat cttccttgct
     3781 aaacgccaaa gaagaggata ttagcattaa taatgaagcc atttccttga tagagtattc
     3841 taggatcgag aaggaactta tagaagcaga tagttctttg aaattacagt gcaacaacct
     3901 cgaactgcta acaagcgtag ctctcgtata cttcgctaag aaaaattgcc aatggtgcat
     3961 aatagaaact ggtttagctg gaaaacagga ccctggaagt ataattgctg gtcaaagtag
     4021 agtctgttgc gccattacta acgtgggcat tagcgatgaa gcttttttat gcaagttttt
     4081 gtctcaaatt actgaaagtt ccacaaataa agcaattttt ctattagacg gttctaatga
     4141 cgaatttgta gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4201 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4261 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     4321 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatat cgctgggcca
     4381 ttctcatgaa gaatatcttg aatttattgt catattacta gttggtgtgg aagtccatat
     4441 atcggtgatc aatatagtgg ttgacatgct ggctagtcaa cattgagcct tttgatcatg
     4501 caaatatatt acggtatttt acaatcaaat atcaaactta actattgact ttataactta
     4561 tttaggtggt aacattctta taaaaaagaa aaaaattact gcaaaacagt actagctttt
     4621 aacttgtatc ctaggttatc tatgctgtct caccatagag aatattacct atttcagaat
     4681 gtatgtccat gattcgccgg gtaaatacat ataatacaca aatctggctt aataaagtct
     4741 ataatatatc tcataaagaa gtgctaaatt ggctagtgct atatattttt aagaaaattt
     4801 cttttgacta agtccatatc gactttgtaa aagttcactt tagcatacat atattacacg
     4861 agccagaaat tgtaactttt gcctaaaatc acaaattgca aaatttaatt gcttgcaaaa
     4921 ggtcacatgc ttataatcaa cttttttaaa aatttaaaat acttttttat tttttatttt
     4981 taaacataaa tgaaataatt tatttattgt ttatgattac cgaaacataa aacctgctca
     5041 agaaaaagaa actgttttgt ccttggaaaa aaagcactac ctaggagcgg ccaaaatgcc
     5101 gaggctttca tagcttaaac tctttacaga aaataggcat tatagatcag ttcgagtttt
     5161 cttattcttc cttccggttt tatcgtcaca gttttacagt aaataagtat cacctcttag
     5221 agttcgatga taagctgtca aacatgagaa ttaattccac atgttaaaat agtgaaggag
     5281 catgttcggc acacagtgga ccgaacgtgg ggtaagtgca ctagggtccg gttaaacgga
     5341 tctcgcattg atgaggcaac gctaattatc aacatataga ttgttatcta tctgcatgaa
     5401 cacgaaatct ttacttgacg acttgaggct gatggtgttt atgcaaagaa accactgtgt
     5461 ttaatatgtg tcactgtttg atattactgt cagcgtagaa gataatagta aaagcggtta
     5521 ataagtgtat ttgagataag tgtgataaag tttttacagc gaaaagacga taaatacaag
     5581 aaaatgatta cgaggatacg gagagaggta tgtacatgtg tatttatata ctaagctgcc
     5641 ggcggttgtt tgcaagaccg agaaaaggct agcaagaatc gggtcattgt agcgtatgcg
     5701 cctgtgaaca ttctcttcaa caagtttgat tccattgcgg tgaaatggta aaagtcaacc
     5761 ccctgcgatg tatattttcc tgtacaatca atcaaaaagc caaatgattt agcattatct
     5821 ttacatcttg ttattttaca gattttatgt ttagatcttt tatgcttgct tttcaaaagg
     5881 cttgcaggca agtgcacaaa caatacttaa ataaatacta ctcagtaata acctatttct
     5941 tagcattttt gacgaaattt gctattttgt tagagtcttt tacaccattt gtctccacac
     6001 ctccgcttac atcaacacca ataacgccat ttaatctaag cgcatcacca acattttctg
     6061 gcgtcagtcc accagctaac ataaaatgta agctctcggg gctctcttgc cttccaaccc
     6121 agtcagaaat cgagttccaa tccaaaagtt cacctgtccc acctgcttct gaatcaaaca
     6181 agggaataaa cgaatgaggt ttctgtgaag ctgcactgag tagtatgttg cagtcttttg
     6241 gaaatacgag tcttttaata actggcaaac cgaggaactc ttggtattct tgccacgact
     6301 catctccatg cagttggacg atatcaatgc cgtaatcatt gaccagagcc aaaacatcct
     6361 ccttaggttg attacgaaac acgccaacca agtatttcgg agtgcctgaa ctatttttat
     6421 atgcttttac aagacttgaa attttccttg caataaccgg gtcaattgtt ctctttctat
     6481 tgggcacaca tataataccc agcaagtcag catcggaatc tagtgcacat tctgcggcct
     6541 ctgtgctctg caagccgcaa actttcacca atggaccaga actacctgtg aaattaataa
     6601 cagacatact ccaagctgcc tttgtgtgct taatcacgta tactcacgtg ctcaatagtc
     6661 accaatgccc tccctcttgg ccctctcctt ttcttttttc gaccgaatta attcttgaag
     6721 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6781 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6901 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6961 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7021 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7081 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7141 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7201 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7261 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7321 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7381 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7441 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7501 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7561 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7621 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7681 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7741 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7801 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7861 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7921 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7981 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8041 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     8101 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8161 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8221 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8281 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     8341 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8401 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8461 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8521 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8581 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     8641 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     8701 agtgagcgag gaagcggaag a
//
