LOCUS       40924_16040        9904 bp DNA     circular SYN 17-DEC-2018
DEFINITION  S. pombe expression vector pDUAL-HFG81 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9904)
  AUTHORS   Matsuyama A, Shirai A, Yashiroda Y, Kamata A, Horinouchi S, Yoshida 
            M.
  TITLE     pDUAL, a multipurpose, multicopy vector capable of chromosomal 
            integration in fission yeast
  JOURNAL   Yeast 21 (15), 1289-1305 (2004)
  PUBMED    15546162
REFERENCE   2  (bases 1 to 9904)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Fission yeast multicopy and integration vector pDUAL-HFG81
  JOURNAL   Published Only in Database (2007)
REFERENCE   3  (bases 1 to 9904)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2007) Akihisa Matsuyama, RIKEN, Chemical Genetics 
            Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan 
            (E-mail:akihisa@riken.jp, URL:http://cgl.riken.go.jp/, 
            Tel:81-48-462-1335, Fax:81-48-462-1336)
REFERENCE   4  (bases 1 to 9904)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 9904)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2004"; volume: "21"; issue: "15"; pages: "1289-1305"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Published 
            Only in Database (2007)"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (15-OCT-2007) Akihisa Matsuyama, RIKEN, Chemical Genetics 
            Laboratory"; volume: " Hirosawa 2-1, Wako, Saitama 351-0198, Japan 
            (E-mail:akihisa@riken.jp, URL:http://cgl.riken.go.jp/, 
            Tel:81-48-462-1335, Fax"; pages: "81-48-462-1336"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     Fission yeast expression vector.
FEATURES             Location/Qualifiers
     source          1..9904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      21..208
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      244..954
                     /pseudo
                     /gene="leu1"
                     /regulatory_class="promoter"
     regulatory      complement(1483..1871)
                     /gene="ura4"
                     /regulatory_class="terminator"
     gene            complement(1483..1871)
                     /gene="ura4"
                     /label=ura4
     CDS             complement(1875..2666)
                     /label=S. pombe ura4
                     /note="orotidine 5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     regulatory      complement(2667..3190)
                     /gene="ura4"
                     /regulatory_class="promoter"
     gene            complement(2667..3190)
                     /gene="ura4"
                     /label=ura4
     rep_origin      complement(3275..4058)
                     /direction=LEFT
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(4781..4797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5010..5465
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5747..5851
                     /label=AmpR promoter
     CDS             5852..6709
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6883..7471
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7759..7780
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7795..7825
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7833..7849
                     /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     7857..7873
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7934..9083
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     CDS             9112..9129
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             9133..9156
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             9160..9873
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
ORIGIN
        1 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
       61 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      121 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
      181 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatatact
      241 agtaagatga ttcttttatc gtcgataacg atgaggatgt agacgattat gatgagagtg
      301 attagcttct aaaatttggt gttttcaaac gctatttaca gtacaatctg aaaaaattgg
      361 atgaacattg taaatggtag gatacgagtt ccaattaatg tattttttaa gtattacact
      421 tagcttatta attttatata tacattctac ctctgtgggt ttaaagacaa ctcgtacaca
      481 tactcaaatt atttatgaac attataatgc atattcgtac tgtctattaa acattttttt
      541 gacattaaaa atttcgttta ctaacgtaga aagcgctgat aattgaggct actattttaa
      601 tatctagaaa caaagattca ttaattaatt aagtaagctc caggatactt gtatatttcg
      661 ttaaaaaagc gcaatttcaa caattcctat gaacatccat gatttcatca caaaatcaaa
      721 catatcagga tttaacgtga attatttgct ttcttgtgat tttcatctga tttctggtca
      781 tttacgttac tgtaaacaag ctttttaaaa aaaggatttt ttacgcaata ttatcgatat
      841 cccaatctgt agtataaaaa gtgcaacagt gttgagtttc ccgaaaccag gcatttttgc
      901 ggtagggtct gaaactttca cgaaaccatt tcacaacaaa acttcttcaa caacatgtgt
      961 gcaaagaaaa tcgtcgtctt accaggagac catattggcc ctgaaattgt tgcttctgcc
     1021 ttggaggttt tgaaagtcgt tgagaagaag cgacctgagt taaaactcga gtttgaagaa
     1081 cacaagattg gaggtgcctc tattgatgcc tatggaaccc ctttgactga cgagactgtg
     1141 aaggcttgtt tggaagctga cggtgttctt ttgggtgccg ttggtggtcc tgaatggacc
     1201 aaccccaatt gtcgtcctga gcaaggttta ttgaagcttc gtaagagtat gggtgtttgg
     1261 gccaaccttc gaccttgcaa ctttgccagc aagtctttag tcaagtacag ccctttgaag
     1321 cctgaaatcg ttgaaggtgt cgatttttgt gttgtacgag aacttactgg aggttgttac
     1381 tttggtgagc gcactgagga caacggatcg ggttatgcta tggacacttg gccttacagt
     1441 ttggaagaag tttctcgtat tgctcgtttg gcggccgcgg gcccaacacc aatgtttata
     1501 accaagtttt atcttgtttg tctacatggt attttacatt catctacata catctttcat
     1561 tggctttgta catagttatc attacaagtc taaaaaaatt cactcttttc ttattcaatg
     1621 tcaatccaag agaaaagatt gtggtaatgt tgtaggagca tgtttaataa attactatag
     1681 caaattactt tttattccca aggtgtttat ctataatagt taatatttta gtcgctacat
     1741 aaaattttac caaagagtac ttgtatacta attctaaatg ccttctgaca taaaacgcct
     1801 aggaaaacaa acgcaaacaa ggcatcgact ttttcaataa ccaaccaaaa aaattttaca
     1861 ttagtctttt tttaatgctg agaaagtctt tgctgatatg ccttccaacc agcttctcta
     1921 tatctcttgg cttcgacaac aggattacga ccagctccat agactccacg accaacaatg
     1981 atgatgtcgc taccgcagtt tacaatcact tcttcaggag tacgatattg ctgtcccagc
     2041 ccgtctcctt taacatccaa gccgatacca ggggacatag ttatgtagtc gctttgaagg
     2101 ttaggaaatc gacgaccagc tataaagcca aagcaaaaat cggtatgctt ctcaaaccat
     2161 tctaaggttt tctctgtgta ggaaccagta gccaaagagc ctttggaaga catttcagcc
     2221 aaaagcaaga gaccacgtcc caaaggtaaa ccaacttctt tgaggccttg tataataccc
     2281 tcgcctggca ctgtatggca atttgtgata tgagcccaag aagcaatttt gtacacacca
     2341 gatgcatatt gtagcttgac ggtatttcca atgtctgcga atttgcgatc ctcaaagata
     2401 agaaaacgat gctttttacc taaggccacc agtttttcta ccatatcctg gtcgaaatcc
     2461 tcgacaacgt caatatgtgt cttgataaca cagacatagg gtccaatttt atctaccaat
     2521 tctaagattt cggatttctt cgtcaaatcg accgcgactg acaagttgct ttgcttttct
     2581 tccatcaaag ccaacaattc cttggcaatg ggatttttca tcccctcagc tctagctgaa
     2641 tagctttgaa atactctagc atccataact ttgcttttaa acctttaatt tcgatccaag
     2701 caaaaaagag gttcttggta ggacaatacg gtaagaaaac acgacatgtg cagagatgcc
     2761 gacgaagcat agttaaactg ggatggtaaa atcaattaag aatttataaa gacaaaattg
     2821 tataagtctc taaaacatct taattatacc tcacagaact atctaaaata tattcacaaa
     2881 gtgcaaacat tatcatgaaa aagaaccatt ttaatttaaa gcaagggcat taaggcttat
     2941 ttacagaatt tcttactttt gtaaagatta taaggctgat tatctttttc accatgccaa
     3001 aaattacaca agatagaatg gatgtttgaa attaaacgtg agtatacaaa caaatacact
     3061 aggtaaatcg aaacattttt ttctccatta agtaacaaat tcctatttag agaaagaatg
     3121 ctgagtagat taaataatct atacaaactt ttttaacaca aatgcataca tatagccagt
     3181 gggatttgta gctaagcttc cgaattcgag tctaactcct taaccactcg gacatagtga
     3241 cttatctgac actcattgta aattaatata tatataggca ttttgtttag ttaaaggtac
     3301 ttaagtaatt agtataaacg aaccaatttt ataatcagga agttaagtga atggtagcac
     3361 atgtcgtaaa aattgtgaat ttttattgaa taatatttta aatacaagcc tttctagact
     3421 aggtatactc ataaacatat atgagcaaaa ggatagagga gattacattg catcttctac
     3481 aaatttattt attgcccttt actgaaaaat taaataatga gtactaaatg ataaaaagcg
     3541 ctcagtacaa gaaagattgc aaaaatattg cattcttcat gaattaaagt tgcatataag
     3601 gcatattgaa agtaatagta ctaaaacagc agttagcgaa aattaataga attatattcg
     3661 caagacaatt gtgacattaa attaaaaaat tgtagaattt ttactatcct ctttaacgcc
     3721 atgagccttt ataaaaaggt taaattagtt ttaacattct ttttttgagt aagagtttac
     3781 aatttatcaa aacctgtgtt attatattca ttagtttcaa tttattagca tctagagaaa
     3841 aatcaattgg cagttacatt gttggaattt atgaagaaaa agaatctaca acggagaata
     3901 agttgctgat cgctttccct aaaattgtat attttgctga gcttattttg acatttcgtt
     3961 gaagttttct ctaattcgca ttcattttaa gtaaaacaat gagaaataaa attacaaaaa
     4021 atacaaatta aaatacaatt tttagctata attatagacg atgcccttgt atcccattct
     4081 gtctcgcttg cccctacttt ttatctttta tataccataa tgaacgctgc cgctactaac
     4141 cataccccga ttttacattt cggactccca aggacgtaca aaatagaaaa ctatagaaaa
     4201 aaataatcag aaaatagcat gtcatctctt tgtaaaacgc gtttgcaaga agaaaggaaa
     4261 caatggagaa gagatcatcc atttgtatgt aaattttagt aaacttgaag aaatcactaa
     4321 caacttctct tacttaggga ttctatgcaa aaccttgtaa atcatctgat ggaggactcg
     4381 atttaatgaa ttggaaggtt gggcccgcgg ccgcttagct gaaacttcca accctcctgc
     4441 tcccgtcaca ttactcgaca aagctaatgt tttggcaact tctcgtcttt ggcgcaagac
     4501 tgtagctaag atttttaagg aggaatatcc tcaccttacc ttaaaaaacc agctcattga
     4561 ctccgcagcc atgcttttgg tcaagagccc tcgtacactt aacggtgttg ttttgactga
     4621 caacttgttt ggtgacatta tttcagatga ggcttctgtc attcctggta gcttgggcct
     4681 tttgccttct gcctcccttt ccggtgtggt aggaaaatca gaagaaaagg ttcattgttt
     4741 ggttgagccc attcacggta gcgctcccga gctcgaattc actggccgtc gttttacaac
     4801 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4861 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4921 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4981 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     5041 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5101 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5161 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5221 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5281 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5341 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5401 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     5461 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5521 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5581 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5641 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5701 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5761 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5821 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5881 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5941 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6001 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6061 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6121 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6181 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6241 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6301 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6361 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6421 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6481 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6541 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6601 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6661 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6721 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6781 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6841 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6901 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6961 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7021 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7081 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7141 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7201 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7261 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7321 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7381 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7441 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7501 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7561 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7621 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7681 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7741 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     7801 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7861 aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgatc gactctagag
     7921 gatcagaaaa ttatcgccat aaaagacaga ataagtcatc agcggttgtt tcatttccta
     7981 tatttttttt ttattttttt attttttaat aagggaaaat ttaacgtcta aggatacaga
     8041 agattgttag cacattaaag taataaaggc ttaagtagta agtgccttag catgttattg
     8101 tatttcaaag gacataatct aaaataataa caatatcatt tctcacaagt tattcaattt
     8161 tctttttttt ttctaataat atcaagaatg tattatttgt ttgacataag tcaactaatt
     8221 tatttaatat gctggattaa tcttgcagac atgtaaatta acaagtttta gtcaaataac
     8281 gttgaagttt caatgaactc aaataatttc tctttttttt tatataactc taatctgatt
     8341 tatattttcc gcagggatca actgaagtta tgacatttgg attggatcac ttataacctt
     8401 ggtcgccaaa taatacaaaa atcagcgtta taaaacaaag aaggtttttg ttaagaaatt
     8461 aatcctcttt cttgataaga aagttgaacc gaaattgcag atactgatat atgaaaataa
     8521 tacccacaat tttgggaata gcgcaagcct caatttaaac aataggtgag gacacatgat
     8581 aatgacctca atgattgtta gaagaaaaga gcctcattac aaaatcgaaa aatgaatggt
     8641 tgggtacaag tttccaaaac atggtaaagt ggactttgcg tatgagacgt aaatagaaaa
     8701 aaacacttgt tatatgtttt ctagaattat tgttgtctct ttatggttgg atgatgcaaa
     8761 atagtaattt cggttagttg ctgtaaaaca ccacgagaca aatagatatg gatatttatt
     8821 aaatcaggaa aaacgtaact ctcggctact ggatggttca gtcacccaac gattactggg
     8881 gagagaaaac agggcaaaag caaagcttaa aggaatccga ttgtcattcg gcaatgtgca
     8941 gcgaaactaa aaaccggata atggacctgt taatcgaaac attgaagaat ggaagaggaa
     9001 tcctggcata tcatcaattg aataagttga attaattatt tcaatctcat tctcactttc
     9061 tgacttatag tcgctttgtt aaagctagcg tcgacaggcc tggatcctat gcatcaccac
     9121 catcatcata tggattataa ggatgatgac gataaaccca tgggtaaagg agaagaactt
     9181 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     9241 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     9301 tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactacttt ctgttatggt
     9361 gttcaatgct tttcaagata cccagatcac atgaaacagc atgacttttt caagagtgcc
     9421 atgcccgaag gctatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     9481 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     9541 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
     9601 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt
     9661 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     9721 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     9781 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct
     9841 gggattacac atggcatgga tgaactatac aaagatatca tgagatctcg actctagagg
     9901 atcc
//
