LOCUS       V001877                 8670 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V001877
VERSION     V001877
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8670)
  AUTHORS   Kobayashi H.
  TITLE     Inducible suppression of global translation by overuse of rare
            codons
  JOURNAL   Appl. Environ. Microbiol. 81 (7), 2544-2553 (2015)
   PUBMED   25636849
REFERENCE   2  (bases 1 to 8670)
  AUTHORS   Kobayashi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JAN-2015) Contact:Hideki Kobayashi Agency for
            Marine-Earth Science and Technology, Institute of Biogeosciences;
            2-15 Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL
            :http://www.jamstec.go.jp/j/
REFERENCE   3  (bases 1 to 8670)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8670)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2015"; volume: "81"; issue: "7"; pages:
            "2544-2553"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (07-JAN-2015) Contact:Hideki Kobayashi Agency for Marine-Earth
            Science and Technology, Institute of Biogeosciences; 2-15
            Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL
            :http://www.jamstec.go.jp/j/"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        152..256
                     /label="AmpR promoter"
     CDS             257..1114
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1288..1876
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2062..2202)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(2903..3195)
                     /label="CEN4"
                     /note="S. cerevisiae CEN4 centromere"
     rep_origin      complement(3196..3649)
                     /direction=LEFT
                     /label="ARS1"
                     /note="S. cerevisiae autonomously replicating sequence
                     ARS1"
     promoter        3651..3866
                     /label="URA3 promoter"
     CDS             3867..4667
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        4750..5191
                     /label="GAL1 promoter"
                     /note="inducible promoter, regulated by Gal4"
     promoter        5223..5241
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5255..5968
                     /label="yeGFP"
                     /note="yeast-enhanced green fluorescent protein"
     terminator      6068..6315
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     CDS             6694..7896
                     /label="AurR"
                     /note="aureobasidin A-resistant mutant of S. cerevisiae
                     inositol phosphorylceramide synthase (Aeed et al., 2009)"
     CDS             complement(8214..8606)
                     /gene="MRP17"
                     /label="Small ribosomal subunit protein bS6m"
                     /note="Small ribosomal subunit protein bS6m from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P28778"
ORIGIN
        1 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
       61 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      121 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      181 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      241 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      301 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      361 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      421 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      481 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      541 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      601 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      661 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      721 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      781 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
      841 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      901 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      961 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1021 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1081 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1141 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1201 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1261 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1321 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1501 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1621 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1681 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1861 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1921 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1981 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2041 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2101 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2161 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2221 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2281 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2341 cagctgggcc attctcatga agaatatctt gaatttattg tcatattact agttggtgtg
     2401 gaagtccata tatcggtgat caatatagtg gttgacatgc tggctagtca acattgagcc
     2461 ttttgatcat gcaaatatat tacggtattt tacaatcaaa tatcaaactt aactattgac
     2521 tttataactt atttaggtgg taacattctt ataaaaaaga aaaaaattac tgcaaaacag
     2581 tactagcttt taacttgtat cctaggttat ctatgctgtc tcaccataga gaatattacc
     2641 tatttcagaa tgtatgtcca tgattcgccg ggtaaataca tataatacac aaatctggct
     2701 taataaagtc tataatatat ctcataaaga agtgctaaat tggctagtgc tatatatttt
     2761 taagaaaatt tcttttgact aagtccatat cgactttgta aaagttcact ttagcataca
     2821 tatattacac gagccagaaa ttgtaacttt tgcctaaaat cacaaattgc aaaatttaat
     2881 tgcttgcaaa aggtcacatg cttataatca acttttttaa aaatttaaaa tactttttta
     2941 ttttttattt ttaaacataa atgaaataat ttatttattg tttatgatta ccgaaacata
     3001 aaacctgctc aagaaaaaga aactgttttg tccttggaaa aaaagcacta cctaggagcg
     3061 gccaaaatgc cgaggctttc atagcttaaa ctctttacag aaaataggca ttatagatca
     3121 gttcgagttt tcttattctt ccttccggtt ttatcgtcac agttttacag taaataagta
     3181 tcacctctta gagttggcgg ttgtttgcaa gaccgagaaa aggctagcaa gaatcgggtc
     3241 attgtagcgt atgcgcctgt gaacattctc ttcaacaagt ttgattccat tgcggtgaaa
     3301 tggtaaaagt caaccccctg cgatgtatat tttcctgtac aatcaatcaa aaagccaaat
     3361 gatttagcat tatctttaca tcttgttatt ttacagattt tatgtttaga tcttttatgc
     3421 ttgcttttca aaaggcctgc aggcaagtgc acaaacaata cttaaataaa tactactcag
     3481 taataaccta tttcttagca tttttgacga aatttgctat tttgttagag tcttttacac
     3541 catttgtctc cacacctccg cttacatcaa caccaataac gccatttaat ctaagcgcat
     3601 caccaacatt ttctggcgtc agtccaccag ctaacataaa atgtaagctt ttcaattcat
     3661 catttttttt ttattctttt ttttgatttc ggtttccttg aaattttttt gattcggtaa
     3721 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     3781 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     3841 aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     3901 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     3961 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     4021 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     4081 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
     4141 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     4201 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     4261 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     4321 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     4381 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     4441 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
     4501 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     4561 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     4621 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     4681 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     4741 tattacccac ggattagaag ccgccgagcg ggtgacagcc ctccgaagga agactctcct
     4801 ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac
     4861 tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg
     4921 gcagtaacct ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga
     4981 taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt
     5041 tgatctatta acagatatat aaatgcaaaa actgcataac cactttaact aatactttca
     5101 acattttcgg tttgtattac ttcttattca aatgtaataa aagtatcaac aaaaaattgt
     5161 taatatacct ctatacttta acgtcaagga gaaaaaaccc cggatcggac tactagcagc
     5221 tgtaatacga ctcactatag ggaatattaa gcttatgtca aagggggagg agctattcac
     5281 aggggtagta cccatactag tagagctaga cggggacgta aatgggcaaa agttctcagt
     5341 atcaggggag ggggaggggg acgctacata cgggaagcta acactaaagt tcatatgtac
     5401 aacagggaag ctacccgtac cctggcccac actagtaaca acattcgggt acggggtaca
     5461 atgtttcgct aggtaccccg accacatgaa gcaacacgac ttcttcaagt cagctatgcc
     5521 cgaggggtac gtacaagaga ggacaatatt ctacaaggac gacgggaatt acaagacaag
     5581 ggctgaggta aagttcgagg gggacacact agtaaatagg atagagctaa aggggataga
     5641 cttcaaggag gacgggaata tactagggca caagatggag tacaattaca attcacacaa
     5701 tgtatacata atggctgaca agcccaagaa tgggataaag gtaaatttca agataaggca
     5761 caatataaag gacgggtcag tacaactagc tgaccactac caacaaaata cacccatagg
     5821 ggacgggccc gtactactac ccgacaatca ctacctatca acacaatcag ctctatcaaa
     5881 ggaccccaat gagaagaggg accacatgat actactagag ttcgtaacag ctgctgggat
     5941 aacacacggg atggacgagc tatacaagta ggagctcgga tccactagta acggccgcca
     6001 gtgtgctgga attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag
     6061 ggccgcatca tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct
     6121 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
     6181 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     6241 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     6301 aaggctttaa tttgccgtac cgagctcgaa tttcgagaaa gtgcccatca gtgttcatat
     6361 ttgttacaaa gtggttttct gatttggtac tactgcagag gcgtattttt tgcttcagtt
     6421 accatagcgt aagaacacta gcgacttttg ttcgtgaacc aacagagtag gatttctact
     6481 gctacatctc ttaggtagtt ggttagtccg atcgctcact tttggttgtt gttaagtact
     6541 tcataagttt atccttttcc tttttcacac tgagctactt tgggtatagc ttttggccca
     6601 aggatctttg aattttctcc aaaagtactt tattttatat cctacaggtt gcggttttca
     6661 tattttaaaa agcttttaat cattcctttg cgtatggcaa accctttttc gagatggttt
     6721 ctatcagaga gacctccaaa ctgccatgta gccgatttag aaacaagttt agatccccat
     6781 caaacgttgt tgaaggtgca aaaatacaaa cccgctttaa gcgactgggt gcattacatc
     6841 ttcttgggat ccatcatgct gtttgtgttc attactaatc ccgcaccttg gatcttcaag
     6901 atcctttttt attgtttctt gggcacttta ttcatcattc cagctacgtc acagtttttc
     6961 ttcaatgcct tgcccatcct aacatgggtg gcgctgtatt tcacttcatc gtactttcca
     7021 gatgaccgca ggcctcctat tactgtcaaa gtgttaccag cggtggaaac aattttatac
     7081 ggcgacaatt taagtgatat tcttgcaaca tcgacgaatt cctttttgga cattttagca
     7141 tggttaccgt acggactatt tcattatggg gccccatttg tcgttgctgc catcttattc
     7201 gtatttggtc caccaactgt tttgcaaggt tatgcttttg catttggtta tatgaacctg
     7261 tttggtgtta tcatgcaaaa tgtctttcca gccgctcccc catggtataa aattctctat
     7321 ggattgcaat cagccaacta tgatatgcat ggctcgcctg gtggattagc tagaattgat
     7381 aagctactcg gtattaatat gtatactaca tgtttttcaa attcctccgt cattttcggt
     7441 gcttttcctt cactgcattc cgggtgtgct actatggaag ccctgttttt ctgttattgt
     7501 tttccaaaat tgaagccctt gtttattgct tatgtttgct ggttatggtg gtcaactatg
     7561 tatctgacac accattattt tgtagacctt atggcaggtt ctgtgctgtc atacgttatt
     7621 ttccagtaca caaagtacac acatttacca attgtagata catctctttt ttgcagatgg
     7681 tcatacactt caattgagaa atacgatata tcaaagagtg atccattggc tgcagattca
     7741 aacgatatcg aaagtgtccc tttgtccaac ttggaacttg actttgatct taatatgact
     7801 gatgaaccca gtgtaagccc ttcgttattt gatggatcta cttctgtttc tcgttcgtcc
     7861 gccacgtcta taacgtcact aggtgtaaag agggcttaat gagtatttta tctgcaatta
     7921 cggatacggt tggtcttatg tagatacata taaatatata tctttttctt tctttttctt
     7981 agtcaggatt gtcgtttagc ataatataca tgtagtttat ttaatcacat accactgatt
     8041 atctttagaa ttttataaat ttttgaaata aatgggtggc ttttaatggt gtctatgtta
     8101 agtgaggctt ttagaatgct cttcctgctt tgtttattat atgtgtatga aagatatgta
     8161 tgtatttaca tgtgtttgta gcgtccccag tcaaaacctg tgcgctatac ctaaatggat
     8221 tgataatctt cattcactaa ttctaaaata gacttcttcc ccaaagaacg gtgtaacgat
     8281 gaggctctat ccagctgctt atctaaatca actttaacga tggatgatct tatgacacgg
     8341 ggatctttct ttaaagttct tagaatttca gactgtaccg cagctgatga atcaaacagc
     8401 attaaaaagt gatatgctcg aaaatgtttt tcctggtctt tcttcattat tttaggaaga
     8461 taccttatgc ccatgggtac aatgtccctc accacacctc tgttttgaat aatcagtttc
     8521 ccgattgtgg aagacaattc ttttgcttcc aactttggcg cattggagtt ggttatgcga
     8581 acaagtccga tcagctcata aagcatctta gtgaaaaggg tggttttgcg ttattctttc
     8641 ctctggatcc tctagagtcg acctcgaggc
//
