LOCUS       sequence-350308-        7707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mWasabi for auxotrophic selection in the absence of 
            methionine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  TITLE     Ralser Yeast Expression plasmids for auxotrophic selection
REFERENCE   2  (bases 1 to 7707)
  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"
COMMENT     Sequence Label: pM-mWasabi
FEATURES             Location/Qualifiers
     source          1..7707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        177..843
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     713..736
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      1610..1797
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1665..1689)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1864..2367
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(2626..3960)
                     /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="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFGNSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENASKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     primer_bind     complement(5599..5618)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5600..5618)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5705..5722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5876..6464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5956..5975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6635..7426,7427..7495))
                     /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     7258..7277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7496..7600)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
       61 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
      121 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgtcat
      181 tatcaatact gccatttcaa agaatacgta aataattaat agtagtgatt ttcctaactt
      241 tatttagtca aaaaattagc cttttaattc tgctgtaacc cgtacatgcc caaaataggg
      301 ggcgggttac acagaatata taacatcgta ggtgtctggg tgaacagttt attcctggca
      361 tccactaaat ataatggagc ccgcttttta agctggcatc cagaaaaaaa aagaatccca
      421 gcaccaaaat attgttttct tcaccaacca tcagttcata ggtccattct cttagcgcaa
      481 ctacagagaa caggggcaca aacaggcaaa aaacgggcac aacctcaatg gagtgatgca
      541 acctgcctgg agtaaatgat gacacaaggc aattgaccca cgcatgtatc tatctcattt
      601 tcttacacct tctattacct tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt
      661 gaaaccagtt ccctgaaatt attcccctac ttgactaata agtatataaa gacggtaggt
      721 attgattgta attctgtaaa tctatttctt aaacttctta aattctactt ttatagttag
      781 tctttttttt agttttaaaa caccaagaac ttagtttcga ataaacacac ataaacaaac
      841 aaagtcgacg gatcggtgac ggtgctggtt taattaacat ggtaagtaaa ggtgaagaga
      901 caactatggg ggtaatcaaa ccagacatga agatcaaact gaaaatggaa ggtaacgtta
      961 atggccacgc gtttgttatt gaaggtgaag gagaggggaa accttacgac ggaaccaaca
     1021 ctatcaattt ggaggttaag gaaggtgcac cattaccatt cagctatgac atcttaacaa
     1081 ctgcattttc atacggcaac agagctttta ccaagtaccc tgacgatata ccaaattact
     1141 tcaaacaatc ctttcctgaa ggttattcat gggagagaac aatgacgttt gaagataagg
     1201 gcattgtcaa ggtcaaatct gacatttcta tggaagagga ttcattcatc tacgagattc
     1261 acctcaaggg cgaaaacttc ccaccaaatg gtccagtgat gcaaaaggaa actactggat
     1321 gggatgcttc tacagaaagg atgtacgtga gagatggagt cttaaaaggt gatgttaaaa
     1381 tgaagttgct acttgaaggt ggtggccatc atagagttga tttcaaaaca atctacagag
     1441 ccaaaaaggc tgtaaaactt cctgattatc attttgtgga tcatcgtata gaaattctaa
     1501 accacgacaa agattacaat aaggtcactg tttatgaaat agcagttgcc agaaattcta
     1561 cagatggaat ggatgaattg tacaaataag gcgcgccact tctaaataag cgaatttctt
     1621 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     1681 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     1741 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     1801 atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc
     1861 accgatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg
     1921 gaatttactc tgtatttatt tatttttatg ttttgtattt ggattttaga aagtaaataa
     1981 agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca
     2041 acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta aatagatata
     2101 cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca
     2161 cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt
     2221 agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
     2281 tctttaattt ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
     2341 attataatta tttttatagc acgtgatgaa aaggacccta agaaaccatt attatcatga
     2401 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggt cgagcttgtg
     2581 agagaaagta ggtttataca taattttaca actcattacg cacactcatg gtttttggcc
     2641 agcgaaaaca gtttcaaaag attgctggaa gtctgcaata atgtcatcaa taaattcgat
     2701 accaacagag acacgaatta agtccttggt aacaccagat gccaactttt ctttgtcatt
     2761 taattgtttg tgggtagtga agtatggagc aatgactaag gtcttggcat caccaacatt
     2821 ggccaagtta gaggcaagct ttaaattgtc aacaacttga gcaccagaaa gtttgaatgg
     2881 gtcagtttcc ttgtcggcat ttggtaagtc ttttacaccg aaagataaga caccaccgaa
     2941 accgttagat agatacttct tagcattttc atgatgagaa tgagatgcta aaccagggta
     3001 tgaaacccaa gatacgtatg gggattgttc taaccatttg gctaacttcg atgcattttc
     3061 accgtgtctt tcagctctca aagataatgt ttcaacacct tgtagtagca agaaagaggc
     3121 aaatgggttc atcaatggac ccaaatctct taatagttca gttctaacat gaacgatgta
     3181 tgccaagtta ccgtaggctt cattgtagat agtaccgtga tatccttcgg caggttgaga
     3241 gaattgaggg aacttttctg ggtagtcctt ccatgggaac ttaccagagt caacaataat
     3301 accaccgata gtagtaccat gaccaccaat ccatttggta gcagaatgtg ttacaatatc
     3361 agcaccgtat ttaattggct gacagaagta accaccggca ccaaatgtgt tgtcaacgac
     3421 aactggaata ccgtgtttgt gagcaattgc aacaattttt tcaaaatccg gaacattgta
     3481 ctttggatta ccaatggttt ccaaataaac agccttggtt ctttcatcaa agaccttttc
     3541 gaattcttct ggattgtcac cttcaacaaa tctagcctcg ataccaaatc ttttgaacga
     3601 gattttgaac tggttataag taccaccgta taagtaagaa gtggaaacga tgttgtcacc
     3661 agtgtgtgcc aaaccttgga tggcaagggt ttgagcggct tgaccggagg aaacagccaa
     3721 agcagcagca ccaccttcta aagcagcaat tctttcttcc aaaacattac tggttgggtt
     3781 ttggaaacgg gaatagacgt aacctggaac ttctagacca aacaattgcg aaccatgctt
     3841 agagtttccg aaaacataag aagtggtggc gtaaattggt acagctctgg atctgtgagc
     3901 attgtcacca gggttctctt ggccggcgtg tagttgaaca gtatcgaaat gagatggcat
     3961 tgtatggatg ggggtaatag aattgtatct atgtatctga cgaccctgta ttacacgaag
     4021 gaaatagtag atgaaaagac aagagagcaa gaaaaaggaa aaacttcctt tccaacagac
     4081 gctttactta accttatata tatcttattt ttttcatcga gcgtgttcaa ttggacaagg
     4141 tgccctgatg aatcccctaa tgattttggt aaaaatcatt aagttaaggt ggatacacat
     4201 cttgtcatat gatcaaatgg tttcgcgaaa aatcaataat cagacaacaa gatgtgcgaa
     4261 ctcgatattt tacacgactc tctttaccaa ttctgccccg aattacactt aaaacgactc
     4321 aacagcttaa cgttggcttg ccacgcatta cttgactgta aaactctcac tcttaccgaa
     4381 cttggccgta acctgccaac caaagcgaga acaaaacata acatcaaacg aatcgaccga
     4441 ttgttaggta atcgtcacct ccacaaagag cgactcgctg tataccgttg gcatgctagc
     4501 tttatctgtt cgggcaatac gatgcccatt gtacttgttg actggtctga tattcgtgag
     4561 caaaaacgac ttatggtatt gcgagcttca gtcgcactac acggtcgttc tgttactctt
     4621 tatgagaaag cgttcccgct ttcagagcaa tgttcaaaga aagctcatga ccaatttcta
     4681 gccgaccttg cgagcattct accgagtaac accacaccgc tcattgtcag tgatgctggc
     4741 tttaaagtgc catggtataa atccgttgag aagctgggtt ggtactggtt aagtcgagta
     4801 agaggaaaag tacaatatgc agacctagga gcggaaaact ggaaacctat cagcaactta
     4861 catgatatgt catctagtca ctcaaagact ttaggctata agaggctgac taaaagcaat
     4921 ccaatctcat gccaaattct attgtataaa tctcgctcta aaggccgaaa aaatcagcgc
     4981 tcgacacgga ctcattgtca ccacccgtca cctaaaatct actcagcgtc ggcaaaggag
     5041 ccatgggttc tagcaactaa cttacctgtt gaaattcgaa cacccaaaca acttgttaat
     5101 atctattcga agcgaatgca gattgaagaa accttccgag acttgaaaag tcctgcctac
     5161 ggactaggcc tacgccatag ccgaacgagc agctcagagc gttttgatat catgctgcta
     5221 atcgccctga tgcttcaact aacatgttgg cttgcgggcg ttcatgctca gaaacaaggt
     5281 tgggacaagc acttccaggc taacacagtc agaaatcgaa acgtactctc aacagttcgc
     5341 ttaggcatgg aagttttgcg gcattctggc tacacaataa caagggaaga cttactcgtg
     5401 gctgcaaccc tactagctca aaatttattc acacatggtt acgctttggg gaaattatga
     5461 ggggatctct cagcaaggtg ccaccttttc gacacttcgg ttattatgtt acacagtttt
     5521 catgaggatg gcggagctcg aattcatcga tgatatcaga tccactagtg gcctatgcgg
     5581 ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc
     5641 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5701 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5761 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5821 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5881 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5941 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6001 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6061 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6121 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6181 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6241 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6301 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6361 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6421 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6481 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6541 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6601 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6661 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6721 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6781 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6841 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6901 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6961 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7021 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7081 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7141 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7201 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7261 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7321 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7381 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7441 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7501 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7561 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7621 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7681 cgaggccctt tcgtctcgcg cgtttcg
//
