LOCUS       40924_16180       10117 bp DNA     circular SYN 17-DEC-2018
DEFINITION  S. pombe expression vector pDUAL2-HFF1c DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10117)
  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 10117)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Fission yeast multicopy and integration vector pDUAL2-HFF1c
  JOURNAL   Published Only in Database (2007)
REFERENCE   3  (bases 1 to 10117)
  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 10117)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 10117)
  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..10117
                     /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"
     rep_origin      975..1758
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(2206..2222)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2435..2890
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3172..3276
                     /label=AmpR promoter
     CDS             3277..4134
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4308..4896
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5184..5205
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5220..5250
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5258..5274
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     5282..5298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(5317..5746)
                     /gene="ura4"
                     /regulatory_class="terminator"
     CDS             complement(5750..6541)
                     /label=S. pombe ura4
                     /note="orotidine 5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     regulatory      complement(6542..7068)
                     /gene="ura4"
                     /regulatory_class="promoter"
     promoter        7117..8272
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces
                     pombe, conferring strong thiamine-repressible expression"
     CDS             8301..8318
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             8322..8345
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             8352..8375
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     protein_bind    8382..8506
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        8531..8561
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             8615..9271
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             9616..9918
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(9962..10086)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
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 agtgcctcta ttgatgccta tggaacccct ttgactgacg agactgtgaa ggcttgtttg
      301 gaagctgacg gtgttctttt gggtgccgtt ggtggtcctg aatggaccaa ccccaattgt
      361 cgtcctgagc aaggtttatt gaagcttcgt aagagtatgg gtgtttgggc caaccttcga
      421 ccttgcaact ttgccagcaa gtctttagtc aagtacagcc ctttgaagcc tgaaatcgtt
      481 gaaggtgtcg atttttgtgt tgtacgagaa cttactggag gttgttactt tggtgagcgc
      541 actgaggaca acggatcggg ttatgctatg gacacttggc cttacagttt ggaagaagtt
      601 tctcgtattg ctcgtttggc ggccgcgggc ccaaccttcc aattcattaa atcgagtcct
      661 ccatcagatg atttacaagg ttttgcatag aatccctaag taagagaagt tgttagtgat
      721 ttcttcaagt ttactaaaat ttacatacaa atggatgatc tcttctccat tgtttccttt
      781 cttcttgcaa acgcgtttta caaagagatg acatgctatt ttctgattat ttttttctat
      841 agttttctat tttgtacgtc cttgggagtc cgaaatgtaa aatcggggta tggttagtag
      901 cggcagcgtt cattatggta tataaaagat aaaaagtagg ggcaagcgag acagaatggg
      961 atacaagggc atcgtctata attatagcta aaaattgtat tttaatttgt attttttgta
     1021 attttatttc tcattgtttt acttaaaatg aatgcgaatt agagaaaact tcaacgaaat
     1081 gtcaaaataa gctcagcaaa atatacaatt ttagggaaag cgatcagcaa cttattctcc
     1141 gttgtagatt ctttttcttc ataaattcca acaatgtaac tgccaattga tttttctcta
     1201 gatgctaata aattgaaact aatgaatata ataacacagg ttttgataaa ttgtaaactc
     1261 ttactcaaaa aaagaatgtt aaaactaatt taaccttttt ataaaggctc atggcgttaa
     1321 agaggatagt aaaaattcta caatttttta atttaatgtc acaattgtct tgcgaatata
     1381 attctattaa ttttcgctaa ctgctgtttt agtactatta ctttcaatat gccttatatg
     1441 caactttaat tcatgaagaa tgcaatattt ttgcaatctt tcttgtactg agcgcttttt
     1501 atcatttagt actcattatt taatttttca gtaaagggca ataaataaat ttgtagaaga
     1561 tgcaatgtaa tctcctctat ccttttgctc atatatgttt atgagtatac ctagtctaga
     1621 aaggcttgta tttaaaatat tattcaataa aaattcacaa tttttacgac atgtgctacc
     1681 attcacttaa cttcctgatt ataaaattgg ttcgtttata ctaattactt aagtaccttt
     1741 aactaaacaa aatgcctata tatatattaa tttacaatga gtgtcagata agtcactatg
     1801 tccgagtggt taaggagtta gactcgggcc cgcggccgct tagctgaaac ttccaaccct
     1861 cctgctcccg tcacattact cgacaaagct aatgttttgg caacttctcg tctttggcgc
     1921 aagactgtag ctaagatttt taaggaggaa tatcctcacc ttaccttaaa aaaccagctc
     1981 attgactccg cagccatgct tttggtcaag agccctcgta cacttaacgg tgttgttttg
     2041 actgacaact tgtttggtga cattatttca gatgaggctt ctgtcattcc tggtagcttg
     2101 ggccttttgc cttctgcctc cctttccggt gtggtaggaa aatcagaaga aaaggttcat
     2161 tgtttggttg agcccattca cggtagcgct cccgagctcg aattcactgg ccgtcgtttt
     2221 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2281 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2341 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2401 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     2461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2641 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2761 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2881 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2941 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3001 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3061 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3121 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3181 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3241 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3301 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3361 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3481 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3661 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3841 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3961 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4141 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4201 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4441 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4921 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4981 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5041 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5101 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5161 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5221 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5281 acaggaaaca gctatgacca tgattacgcc aagcttgtga tattgacgaa actttttgac
     5341 atctaattta ttctgttcca acaccaatgt ttataaccaa gttttatctt gtttgtctac
     5401 atggtatttt acattcatct acatacatct ttcattggct ttgtacatag ttatcattac
     5461 aagtctaaaa aaattcactc ttttcttatt caatgtcaat ccaagagaaa agattgtggt
     5521 aatgttgtag gagcatgttt aataaattac tatagcaaat tactttttat tcccaaggtg
     5581 tttatctata atagttaata ttttagtcgc tacataaaat tttaccaaag agtacttgta
     5641 tactaattct aaatgccttc tgacataaaa cgcctaggaa aacaaacgca aacaaggcat
     5701 cgactttttc aataaccaac caaaaaaatt ttacattagt ctttttttaa tgctgagaaa
     5761 gtctttgctg atatgccttc caaccagctt ctctatatct cttggcttcg acaacaggat
     5821 tacgaccagc tccatagact ccacgaccaa caatgatgat gtcgctaccg cagtttacaa
     5881 tcacttcttc aggagtacga tattgctgtc ccagcccgtc tcctttaaca tccaagccga
     5941 taccagggga catagttatg tagtcgcttt gaaggttagg aaatcgacga ccagctataa
     6001 agccaaagca aaaatcggta tgcttctcaa accattctaa ggttttctct gtgtaggaac
     6061 cagtagccaa agagcctttg gaagacattt cagccaaaag caagagacca cgtcccaaag
     6121 gtaaaccaac ttctttgagg ccttgtataa taccctcgcc tggcactgta tggcaatttg
     6181 tgatatgagc ccaagaagca attttgtaca caccagatgc atattgtagc ttgacggtat
     6241 ttccaatgtc tgcgaatttg cgatcctcaa agataagaaa acgatgcttt ttacctaagg
     6301 ccaccagttt ttctaccata tcctggtcga aatcctcgac aacgtcaata tgtgtcttga
     6361 taacacagac atagggtcca attttatcta ccaattctaa gatttcggat ttcttcgtca
     6421 aatcgaccgc gactgacaag ttgctttgct tttcttccat caaagccaac aattccttgg
     6481 caatgggatt tttcatcccc tcagctctag ctgaatagct ttgaaatact ctagcatcca
     6541 taactttgct tttaaacctt taatttcgat ccaagcaaaa aagaggttct tggtaggaca
     6601 atacggtaag aaaacacgac atgtgcagag atgccgacga agcatagtta aactgggatg
     6661 gtaaaatcaa ttaagaattt ataaagacaa aattgtataa gtctctaaaa catcttaatt
     6721 atacctcaca gaactatcta aaatatattc acaaagtgca aacattatca tgaaaaagaa
     6781 ccattttaat ttaaagcaag ggcattaagg cttatttaca gaatttctta cttttgtaaa
     6841 gattataagg ctgattatct ttttcaccat gccaaaaatt acacaagata gaatggatgt
     6901 ttgaaattaa acgtgagtat acaaacaaat acactaggta aatcgaaaca tttttttctc
     6961 cattaagtaa caaattccta tttagagaaa gaatgctgag tagattaaat aatctataca
     7021 aactttttta acacaaatgc atacatatag ccagtgggat ttgtagctaa gcttgcatgc
     7081 ctgcaggtcg atcgactcta gaggatcaga aaattatcgc cataaaagac agaataagtc
     7141 atcagcggtt gtttcatttc ctatattttt tttttatttt tttatttttt aataagggaa
     7201 aatttaacgt ctaaggatac agaagattgt tagcacatta aagtaataaa ggcttaagta
     7261 gtaagtgcct tagcatgtta ttgtatttca aaggacataa tctaaaataa taacaatatc
     7321 atttctcaca agttattcaa ttttcttttt tttttctaat aatatcaaga atgtattatt
     7381 tgtttgacat aagtcaacta atttatttaa tatgctggat taatcttgca gacatgtaaa
     7441 ttaacaagtt ttagtcaaat aacgttgaag tttcaatgaa ctcaaataat ttctcttttt
     7501 ttttatataa ctctaatctg atttatattt tccgcaggga tcaactgaag ttatgacatt
     7561 tggattggat cacttataac cttggtcgcc aaataataca aaaatcagcg ttataaaaca
     7621 aagaaggttt ttgttaagaa attaatcctc tttcttgata agaaagttga accgaaattg
     7681 cagatactga tatatgaaaa taatacccac aattttggga atagcgcaag cctcaattta
     7741 aacaataggt gaggacacat gataatgacc tcaatgattg ttagaagaaa agagcctcat
     7801 tacaaaatcg aaaaatgaat ggttgggtac aagtttccaa aacatggtaa agtggacttt
     7861 gcgtatgaga cgtaaataga aaaaaacact tgttatatgt tttctagaat tattgttgtc
     7921 tctttatggt tggatgatgc aaaatagtaa tttcggttag ttgctgtaaa acaccacgag
     7981 acaaatagat atggatattt attaaatcag gaaaaacgta actctcggct actggatggt
     8041 tcagtcaccc aacgattact ggggagagaa aacagggcaa aagcaaagct taaaggaatc
     8101 cgattgtcat tcggcaatgt gcagcgaaac taaaaaccgg ataatggacc tgttaatcga
     8161 aacattgaag atatataaag gaagaggaat cctggcatat catcaattga ataagttgaa
     8221 ttaattattt caatctcatt ctcactttct gacttatagt cgctttgtta aagctagcgt
     8281 cgacaggcct ggatcctatg catcaccacc atcatcatat ggattataag gatgatgacg
     8341 ataaacccat ggactacaag gacgacgatg acaaagatat cacaagtttg tacaaaaaag
     8401 ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa
     8461 aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg ccgcattagg
     8521 caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt tgagttagga
     8581 tccggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     8641 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     8701 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     8761 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     8821 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     8881 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     8941 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     9001 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     9061 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     9121 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     9181 tcaggttcat catgccgtct gtgatggctt ccatgtcggc agaatgctta atgaattaca
     9241 acagtactgc gatgagtggc agggcggggc gtaaacgcgt ggatccggct tactaaaagc
     9301 cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat
     9361 gtatacccga agtatgtcaa aaagaggtgt gctatgaagc agcgtattac agtgacagtt
     9421 gacagcgaca gctatcagtt gctcaaggca tatatgatgt caatatctcc ggtctggtaa
     9481 gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc
     9541 aggaagggat ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gctgacgaga
     9601 acagggactg gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt
     9661 ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc
     9721 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat
     9781 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
     9841 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
     9901 ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc tgcaggtcga
     9961 ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
    10021 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt tcttgtacaa
    10081 agtggtgata tcatgagatc tcgactctag aggatcc
//
