LOCUS       40924_38068        8009 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS435PGK DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8009)
  AUTHORS   Mumberg D, Muller R, Funk M.
  TITLE     Yeast vectors for the controlled expression of heterologous proteins
            in different genetic backgrounds
  JOURNAL   Gene 156 (1), 119-122 (1995)
  PUBMED    7737504
REFERENCE   2  (bases 1 to 8009)
  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   3  (bases 1 to 8009)
  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   4  (bases 1 to 8009)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8009)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1995"; volume: "156"; issue: "1"; pages: "119-122"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2009"; volume: "82"; issue: "5"; 
            pages: "837-845"
COMMENT     SGRef: number: 3; 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: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(666..1757)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(1770..2174)
                     /label=LEU2 promoter
     rep_origin      2866..3746
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     4610..4626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4636..4654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(4679..4926)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4936..4952
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4986..5002)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(5013..5731)
                     /gene="PGK1"
                     /regulatory_class="promoter"
     gene            complement(5013..5731)
                     /gene="PGK1"
                     /label=PGK1
     promoter        complement(5749..5767)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5788..5804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5812..5828)
                     /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(5836..5866)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5881..5902)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6952..7809)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7810..7914)
                     /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 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatgcttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaagattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accagccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc catttgctta tacaatcttc
     2821 ctgtttttgg ggcttttctg attatcaacc ggggtggagc ttcccattgc gaataccgct
     2881 tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat atccctatat
     2941 aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct ctacattttt
     3001 tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac tattcataga
     3061 gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat agagacaaaa
     3121 tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta tcactttctg
     3181 ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc ctttatcttg
     3241 aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga gtcaggcttt
     3301 ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac ggacctacag
     3361 tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa aagtaatcta
     3421 agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca aaaaagaagt
     3481 atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg taaaaatgca
     3541 gctcagattc tttgtttgaa aaattagcgc tctcgtcgcg ttgcattttt gttttacaaa
     3601 aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt ctgttctgta
     3661 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgttct
     3721 acaaaatgaa gcacagatgc ttcgttaaca aagatatgct attgaagtgc aagatggaaa
     3781 cgcagaaaat gaaccgggga tgcgacgtgc aagattacct atgcaataga tgcaatagtt
     3841 tctccaggaa ccgaaataca tacattgtct tccgtaaagc gctagactat atattattat
     3901 acaggttcaa atatactatc tgtttcaggg aaaactccca ggttcggatg ttcaaaattc
     3961 aatgatgggt aacaagtacg atcgtaaatc tgtaaaacag tttgtcggat attaggctgt
     4021 atctcctcaa agcgtattcg aatatcattg agaagctgca gcgtcacatc ggataataat
     4081 gatggcagcc attgtagaag tgccttttgc atttctagtc tctttctcgg tctagctagt
     4141 tttactacat cgcgaagata gaatcttaga tcacactgcc tttgctgagc tggatcaata
     4201 gagtaacaaa agagtggtaa ggcctcgtta aaggacaagg acctgagcgg aagtgtatcg
     4261 tacagtagac ggagtatcta gtatagtcta tagtccgtgg aattaattct catctttgac
     4321 agcttatcat cgataagggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4381 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4441 cttaatgcgc cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg caactgttgg
     4501 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4561 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4621 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gcccggccgc
     4681 aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac
     4741 atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
     4801 atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
     4861 ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt
     4921 acatgatgcg gccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc
     4981 cgggggatcc actagttcta gagcggccgc caccgcggtg ttttatattt gttgtaaaaa
     5041 gtagataatt acttccttga tgatctgtaa aaaagagaaa aagaaagcat ctaagaactt
     5101 gaaaaactac gaattagaaa agaccaaata tgtatttctt gcattgacca atttatgcaa
     5161 gtttatatat atgtaaatgt aagtttcacg aggttctact aaactaaacc acccccttgg
     5221 ttagaagaaa agagtgtgtg agaacaggct gttgttgtca cacgattcgg acaattctgt
     5281 ttgaaagaga gagagtaaca gtacgatcga acgaactttg ctctggagat cacagtgggc
     5341 atcatagcat gtggtactaa accctttccc gccattccag aaccttcgat tgcttgttac
     5401 aaaacctgtg agccgtcgct aggaccttgt tgtgtgacga aattggaagc tgcaatcaat
     5461 aggaagacag gaagtcgagc gtgtctgggt tttttcagtt ttgttctttt tgcaaacaaa
     5521 tcacgagcga cggtaatttc tttctcgata agaggccacg tgctttatga gggtaacatc
     5581 aattcaagaa ggagggaaac acttcctttt tctggccctg ataatagtat gagggtgaag
     5641 ccaaaataaa ggattcgcgc ccaaatcggc atctttaaat gcaggtatgc gatagttcct
     5701 cactctttcc ttactcacga gtaattcttg gagctccagc ttttgttccc tttagtgagg
     5761 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5821 gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta
     5881 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7921 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7981 aaataggcgt atcacgaggc cctttcgtc
//
