LOCUS       40924_38023        6926 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS433GAP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  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 6926)
  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 6926)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6926)
  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..6926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        317..503
                     /label=HIS3 promoter
     CDS             504..1160
                     /codon_start=1
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     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"
     promoter        complement(3968..4634)
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        complement(4666..4684)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4705..4721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4729..4745)
                     /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(4753..4783)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4798..4819)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5107..5695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5869..6726)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6727..6831)
                     /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 accgcggttt gtttgtttat gtgtgtttat tcgaaactaa gttcttggtg ttttaaaact
     4021 aaaaaaaaga ctaactataa aagtagaatt taagaagttt aagaaataga tttacagaat
     4081 tacaatcaat acctaccgtc tttatatact tattagtcaa gtaggggaat aatttcaggg
     4141 aactggtttc aacctttttt ttcagctttt tccaaatcag agagagcaga aggtaataga
     4201 aggtgtaaga aaatgagata gatacatgcg tgggtcaatt gccttgtgtc atcatttact
     4261 ccaggcaggt tgcatcactc cattgaggtt gtgcccgttt tttgcctgtt tgtgcccctg
     4321 ttctctgtag ttgcgctaag agaatggacc tatgaactga tggttggtga agaaaacaat
     4381 attttggtgc tgggattctt tttttttctg gatgccagct taaaaagcgg gctccattat
     4441 atttagtgga tgccaggaat aaactgttca cccagacacc tacgatgtta tatattctgt
     4501 gtaacccgcc ccctattttg ggcatgtacg ggttacagca gaattaaaag gctaattttt
     4561 tgactaaata aagttaggaa aatcactact attaattatt tacgtattct ttgaaatggc
     4621 agtattgata atgataaact cgaactggag ctccagcttt tgttcccttt agtgagggtt
     4681 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4741 cacaattcca cacaacatag gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4801 agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4861 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5761 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5821 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5881 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5941 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6001 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6061 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6121 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6181 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6241 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6301 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6361 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6421 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6481 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6541 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6601 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6661 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6721 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6781 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6841 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6901 taggcgtatc acgaggccct ttcgtc
//
