LOCUS       40924_38143        6848 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRS446GAP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6848)
  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 6848)
  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 6848)
  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 6848)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6848)
  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..6848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..416
                     /label=URA3 promoter
     CDS             417..1217
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(2270..3150)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     3487..3503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3513..3531
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(3550..3797)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3807..3823
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3857..3873)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3890..4556)
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        complement(4588..4606)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4627..4643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4651..4667)
                     /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(4675..4705)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4720..4741)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5029..5617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5791..6648)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6649..6753)
                     /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 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccct
     1681 tatcgatgat aagctgtcaa agatgagaat taattccacg gactatagac tatactagat
     1741 actccgtcta ctgtacgata cacttccgct caggtccttg tcctttaacg aggccttacc
     1801 actcttttgt tactctattg atccagctca gcaaaggcag tgtgatctaa gattctatct
     1861 tcgcgatgta gtaaaactag ctagaccgag aaagagacta gaaatgcaaa aggcacttct
     1921 acaatggctg ccatcattat tatccgatgt gacgctgcag cttctcaatg atattcgaat
     1981 acgctttgag gagatacagc ctaatatccg acaaactgtt ttacagattt acgatcgtac
     2041 ttgttaccca tcattgaatt ttgaacatcc gaacctggga gttttccctg aaacagatag
     2101 tatatttgaa cctgtataat aatatatagt ctagcgcttt acggaagaca atgtatgtat
     2161 ttcggttcct ggagaaacta ttgcatctat tgcataggta atcttgcacg tcgcatcccc
     2221 ggttcatttt ctgcgtttcc atcttgcact tcaatagcat atctttgtta acgaagcatc
     2281 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     2341 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     2401 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga cgagagcgct aatttttcaa
     2461 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     2521 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     2581 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     2641 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     2701 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     2761 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     2821 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     2881 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     2941 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     3001 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     3061 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     3121 agcaatgttt gtggaagcgg tattcgcaat gggaagctcc accccggttg ataatcagaa
     3181 aagccccaaa aacaggaaga ttgtataagc aaatggcgaa cgtggcgaga aaggaaggga
     3241 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3301 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcgcgccat tcgccattca
     3361 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3421 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3481 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
     3541 gtaccggccg caaattaaag ccttcgagcg tcccaaaacc ttctcaagca aggttttcag
     3601 tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa
     3661 taacgttctt aatactaaca taactataaa aaaataaata gggacctaga cttcaggttg
     3721 tctaactcct tccttttcgg ttagagcgga tgtgggggga gggcgtgaat gtaagcgtga
     3781 cataactaat tacatgatgc ggccctcgag gtcgacggta tcgataagct tgatatcgaa
     3841 ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt ttgtttgttt
     3901 atgtgtgttt attcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat
     3961 aaaagtagaa tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg
     4021 tctttatata cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt
     4081 ttttcagctt tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga
     4141 tagatacatg cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac
     4201 tccattgagg ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta
     4261 agagaatgga cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc
     4321 tttttttttc tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga
     4381 ataaactgtt cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt
     4441 tgggcatgta cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg
     4501 aaaatcacta ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa
     4561 ctcgaactgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     4621 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4681 aggagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt
     4741 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4801 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5461 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5641 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5701 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5761 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5821 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5881 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5941 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6001 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6061 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6121 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6181 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6241 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6301 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6361 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6421 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6481 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6541 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6601 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6721 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6781 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6841 ctttcgtc
//
