LOCUS       40924_38123        7977 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS445GAP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7977)
  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 7977)
  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 7977)
  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 7977)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7977)
  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..7977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(666..1757)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /label=LEU2 promoter
     rep_origin      complement(3393..4273)
                     /direction=LEFT
                     /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"
     promoter        complement(5019..5685)
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        complement(5717..5735)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5756..5772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5780..5796)
                     /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(5804..5834)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5849..5870)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6158..6746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6920..7777)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7778..7882)
                     /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 ccttatcgat gataagctgt
     2821 caaagatgag aattaattcc acggactata gactatacta gatactccgt ctactgtacg
     2881 atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt tgttactcta
     2941 ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac
     3001 tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat
     3061 tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac
     3121 agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga
     3181 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     3241 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     3301 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     3361 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     3421 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     3481 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     3541 tgtaaaacaa aaatgcaacg cgacgagagc gctaattttt caaacaaaga atctgagctg
     3601 catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac
     3661 ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta
     3721 gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact
     3781 gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa
     3841 agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca
     3901 agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca
     3961 gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt
     4021 ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc
     4081 tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa
     4141 aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat
     4201 atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag
     4261 cggtattcgc aatgggaagc tccaccccgg ttgataatca gaaaagcccc aaaaacagga
     4321 agattgtata agcaaatggc 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 caccgcggtt tgtttgttta tgtgtgttta
     5041 ttcgaaacta agttcttggt gttttaaaac taaaaaaaag actaactata aaagtagaat
     5101 ttaagaagtt taagaaatag atttacagaa ttacaatcaa tacctaccgt ctttatatac
     5161 ttattagtca agtaggggaa taatttcagg gaactggttt caaccttttt tttcagcttt
     5221 ttccaaatca gagagagcag aaggtaatag aaggtgtaag aaaatgagat agatacatgc
     5281 gtgggtcaat tgccttgtgt catcatttac tccaggcagg ttgcatcact ccattgaggt
     5341 tgtgcccgtt ttttgcctgt ttgtgcccct gttctctgta gttgcgctaa gagaatggac
     5401 ctatgaactg atggttggtg aagaaaacaa tattttggtg ctgggattct ttttttttct
     5461 ggatgccagc ttaaaaagcg ggctccatta tatttagtgg atgccaggaa taaactgttc
     5521 acccagacac ctacgatgtt atatattctg tgtaacccgc cccctatttt gggcatgtac
     5581 gggttacagc agaattaaaa ggctaatttt ttgactaaat aaagttagga aaatcactac
     5641 tattaattat ttacgtattc tttgaaatgg cagtattgat aatgataaac tcgaactgga
     5701 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     5761 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa
     5821 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
     5881 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7921 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
