LOCUS       sequence-108559-        6727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible MET15 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6727)
  AUTHORS   Gnugge R, Liphardt T, Rudolf F
  TITLE     A shuttle vector series for precise genetic engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2015 Dec 9. doi: 10.1002/yea.3144.
  PUBMED    26647923
REFERENCE   2  (bases 1 to 6727)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast. 2015
            Dec 9. doi: 10.1002/yea.3144."
COMMENT     Sequence Label: FRP1671
FEATURES             Location/Qualifiers
     source          1..6727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..521
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(839..1642)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDLLTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSACVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1643..1859)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     1910..1932
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1924..1941
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1925..1941
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1948..1967
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1948..1966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2384..3718
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     promoter        complement(4410..4428)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4449..4465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4449..4465)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4462..4484)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4473..4489
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     complement(4799..4816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4970..5558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5050..5069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5737..6528,6529..6597))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6360..6379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6598..6702)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
       61 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
      121 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
      181 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
      241 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
      301 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
      361 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
      421 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
      481 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtcttaat taactctggc
      541 gcgccactac tttttggaac tcttgttgtt ctttggagtt caatgcgtcc atctttacag
      601 tcctgtctta ttgttcttga tttgtgcccc gtaaaatact gttacttggt tctggcgagg
      661 tattggatag ttccttttta taaaggccat gaaggttttt ctttccaatt tttttttttt
      721 cgtcattata gaaatcatta cgaccgagat tcccggcata tggtaataac tgatataatt
      781 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
      841 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
      901 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
      961 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     1021 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     1081 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     1141 tacccttagt atattctcca gtagctaggg agcccttgca tgacaattct gctaacatca
     1201 aaaggcctct aggttccttt gttacttctt ccgccgcctg cttcaaaccg ctaacaatac
     1261 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     1321 acgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     1381 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     1441 aatcagtcaa gagatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     1501 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     1561 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     1621 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcacgttttt gttctgtgca
     1681 gttgggttaa gaatactggg caatttcatg tttcttcaac accacctatg cgtatatata
     1741 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ctcggagatt accgaatcaa
     1801 aaaaatttca aagaaaccgg aatcaaaaaa aagaacaaaa aaaaaaaaga tgaatgaaaa
     1861 gcgatcggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     1921 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attggagaaa
     1981 cagaataata tataaacctg tataatataa ccttgaagac tatatttctt ttcgcggccg
     2041 cctatagggc gaattggagc tccctaggtg ctgtatatac tcacagcata actgtatata
     2101 cacccagggt ctaggtgctg tatatactca cagcataact gtatatacac ccagggtcta
     2161 ggtgctgtat atactcacag cataactgta tatacaccca gggtctaggt gctgtatata
     2221 ctcacagcat aactgtatat acacccaggg tctagagcat gtgctctgta tgtatataaa
     2281 actcttgttt tcttcttttc tcttctttcc ttatacatta ggacctttgc agcataaatt
     2341 actatacttc tatactagtg gatcccccgg gctgcaggaa ttcatgccat ctcatttcga
     2401 tactgttcaa ctacacgccg gtcaagagaa ccctggtgac aatgctcaca gatccagagc
     2461 tgtaccaatt tacgccacca cttcttatgt tttcgaaaac tctaagcatg gttcgcaatt
     2521 gtttggttta gaagttccag gttacgtcta ttcccgtttc caaaacccaa ccagtaatgt
     2581 tttggaagaa agaattgctg ctttagaagg tggtgctgct gctttggctg tttcctccgg
     2641 tcaagccgct caaacccttg ccatccaagg tttggcacac actggtgaca acatcgtttc
     2701 cacttcttac ttatacggtg gtacttataa ccagttcaaa atctcgttca aaagatttgg
     2761 tatcgaggct agatttgttg aaggtgacaa tccagaagaa tttgaaaagg tctttgatga
     2821 aagaaccaag gctgtttatt tggaaaccat tggtaatcca aagtacaatg ttccggattt
     2881 tgaaaaaatt gttgcaattg ctcacaaaca cggtattcca gttgtcgttg acaacacatt
     2941 tggtgccggt ggttacttct gtcagccaat taaatacggt gctgatattg taacacattc
     3001 tgctaccaaa tggattggtg gtcatggtac tactatcggt ggtattattg ttgactctgg
     3061 taagttccca tggaaggact acccagaaaa gttccctcaa ttctctcaac ctgccgaagg
     3121 ttatcacggt actatctaca atgaagccta cggtaacttg gcatacatcg ttcatgttag
     3181 aactgaacta ttaagagatt tgggtccatt gatgaaccca tttgcctctt tcttgctact
     3241 acaaggtgtt gaaacattat ctttgagagc tgaaagacac ggtgaaaatg cattgaagtt
     3301 agccaaatgg ttagaacaat ccccatacgt atcttgggtt tcataccctg gtttagcatc
     3361 tcattctcat catgaaaatg ctaagaagta tctatctaac ggtttcggtg gtgtcttatc
     3421 tttcggtgta aaagacttac caaatgccga caaggaaact gacccattca aactttctgg
     3481 tgctcaagtt gttgacaatt taaagctagc ctctaacttg gccaatgttg gtgatgccaa
     3541 gaccttagtc attgctccat acttcactac ccacaaacaa ttaaatgaca aagaaaagtt
     3601 ggcatctggt gttaccaagg acttaattcg tgtctctgtt ggtatcgaat ttattgatga
     3661 cattattgca gacttccagc aatcttttga aactgttttc gctggccaaa aaccatgagt
     3721 gtgcgtaatg agttgtaaaa ttatgtataa acctactttc tctcacaagt actatacttt
     3781 tataaaacga actttattga aatgaatatc ctttttttcc cttgttacat gtcgtgactc
     3841 gtactttgaa cctaaattgt tctaacatca aagaacagtg ttaattcgca gtcgagaaga
     3901 aaaatatggt gaacaagact catctacttc atgagactac tttacgcctc ctataaagct
     3961 gtcacactgg ataaatttat tgtaggacca agttacaaaa gaggatgatg gaggtttctt
     4021 tacaataaag aagcacatgt gtgttaacgt ttttagtatt tgcttgttat gtaaatcagg
     4081 aaaacttcgc gggatttggt tggatgctac tttccataca ataaatatta tagatcaaaa
     4141 aagccaaatt acaagtaaag attagtaaag ctgttgcaat tccatcgttg ataaaaatgt
     4201 tagttattaa atataaaagt cagaataggt gaacttggat ttaattgttg gcatttcgtt
     4261 gctgctagag gccataatat tagatagcca ggacatacta ggtctcctcg tggtatagga
     4321 atggtaccca gcttcggccg tatttacaga taacgatgtc gttattatat atatatatat
     4381 atattgtcac ttgtacccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
     4441 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaac agctggatga
     4501 attgaatagc tttatggacc ctgaaaccac agccacatta accttctttg atggtcaaaa
     4561 cttatccttc accataaata tgcctcgcaa aaaaggtaat taacatatat agaattacat
     4621 tatttatgaa atatcatcac tatctcttag catctttaat ccttttctac atcagataac
     4681 ttcggtttgt tatcatcgtc tgtattgtca tcaattggcg cgcctctccc tgcaggcatt
     4741 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4801 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4861 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttgtt
     5581 taaacctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5641 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5701 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5821 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5881 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5941 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6001 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6061 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6121 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6181 ggttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6241 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6301 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6361 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6421 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6481 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6541 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6601 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6661 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6721 gccacct
//
