LOCUS       40924_38028        6958 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS433PGK DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  AUTHORS   Ohto C, Muramatsu M, Obata S, Sakuradani E, Shimizu S.
  TITLE     Overexpression of the gene encoding HMG-CoA reductase in 
            Saccharomyces cerevisiae for production of prenyl alcohols
  JOURNAL   Appl. Microbiol. Biotechnol. 82 (5), 837-845 (2009)
  PUBMED    19083230
REFERENCE   2  (bases 1 to 6958)
  AUTHORS   Ohto C.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2007) Contact:Chikara Ohto Toyota Motor 
            Corporation, Bio Research Laboratory, Future Project Division; 1, 
            Toyota-cho, Toyota, Aichi 471-8572, Japan
REFERENCE   3  (bases 1 to 6958)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6958)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2009"; volume: "82"; issue: "5"; 
            pages: "837-845"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-MAY-2007) Contact:Chikara Ohto Toyota Motor Corporation, Bio 
            Research Laboratory, Future Project Division; 1, Toyota-cho, Toyota,
            Aichi 471-8572, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        317..503
                     /label=HIS3 promoter
     CDS             504..1160
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     rep_origin      1815..2695
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     3559..3575
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3585..3603
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(3628..3875)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3885..3901
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3935..3951)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3962..4680)
                     /gene="PGK1"
                     /regulatory_class="promoter"
     gene            complement(3962..4680)
                     /gene="PGK1"
                     /label=PGK1
     promoter        complement(4698..4716)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4737..4753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4761..4777)
                     /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)."
     promoter        complement(4785..4815)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4830..4851)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5139..5727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5901..6758)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6759..6863)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg
     1801 gggtggagct tcccattgcg aataccgctt ccacaaacat tgctcaaaag tatctctttg
     1861 ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg ctccttgaac
     1921 ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac tctttagaca
     1981 aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa aatgtaaaca
     2041 tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt ttctgaccaa
     2101 tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc acatcggtat
     2161 agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc gctagtaatc
     2221 agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga caccaaagta
     2281 gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac tgcattatag
     2341 agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata gcgctctcgg
     2401 gatgcatttt tgtagaacaa aaaagaagta tagattcttt gttggtaaaa tagcgctctc
     2461 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     2521 ctcgtcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     2581 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     2641 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttaacaa
     2701 agatatgcta ttgaagtgca agatggaaac gcagaaaatg aaccggggat gcgacgtgca
     2761 agattaccta tgcaatagat gcaatagttt ctccaggaac cgaaatacat acattgtctt
     2821 ccgtaaagcg ctagactata tattattata caggttcaaa tatactatct gtttcaggga
     2881 aaactcccag gttcggatgt tcaaaattca atgatgggta acaagtacga tcgtaaatct
     2941 gtaaaacagt ttgtcggata ttaggctgta tctcctcaaa gcgtattcga atatcattga
     3001 gaagctgcag cgtcacatcg gataataatg atggcagcca ttgtagaagt gccttttgca
     3061 tttctagtct ctttctcggt ctagctagtt ttactacatc gcgaagatag aatcttagat
     3121 cacactgcct ttgctgagct ggatcaatag agtaacaaaa gagtggtaag gcctcgttaa
     3181 aggacaagga cctgagcgga agtgtatcgt acagtagacg gagtatctag tatagtctat
     3241 agtccgtgga attaattctc atctttgaca gcttatcatc gataagggcg aacgtggcga
     3301 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3361 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgcc
     3421 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     3481 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     3541 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     3601 agggcgaatt gggtaccggg cccggccgca aattaaagcc ttcgagcgtc ccaaaacctt
     3661 ctcaagcaag gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag
     3721 aaaaatttga aatataaata acgttcttaa tactaacata actataaaaa aataaatagg
     3781 gacctagact tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg
     3841 gcgtgaatgt aagcgtgaca taactaatta catgatgcgg ccctcgaggt cgacggtatc
     3901 gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgcc
     3961 accgcggtgt tttatatttg ttgtaaaaag tagataatta cttccttgat gatctgtaaa
     4021 aaagagaaaa agaaagcatc taagaacttg aaaaactacg aattagaaaa gaccaaatat
     4081 gtatttcttg cattgaccaa tttatgcaag tttatatata tgtaaatgta agtttcacga
     4141 ggttctacta aactaaacca cccccttggt tagaagaaaa gagtgtgtga gaacaggctg
     4201 ttgttgtcac acgattcgga caattctgtt tgaaagagag agagtaacag tacgatcgaa
     4261 cgaactttgc tctggagatc acagtgggca tcatagcatg tggtactaaa ccctttcccg
     4321 ccattccaga accttcgatt gcttgttaca aaacctgtga gccgtcgcta ggaccttgtt
     4381 gtgtgacgaa attggaagct gcaatcaata ggaagacagg aagtcgagcg tgtctgggtt
     4441 ttttcagttt tgttcttttt gcaaacaaat cacgagcgac ggtaatttct ttctcgataa
     4501 gaggccacgt gctttatgag ggtaacatca attcaagaag gagggaaaca cttccttttt
     4561 ctggccctga taatagtatg agggtgaagc caaaataaag gattcgcgcc caaatcggca
     4621 tctttaaatg caggtatgcg atagttcctc actctttcct tactcacgag taattcttgg
     4681 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     4741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat aggagccgga
     4801 agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt aattgcgttg
     4861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5941 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6001 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6061 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6901 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
