LOCUS       sequence-350987-        6360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TagRFP657 for auxotrophic selection in the absence of 
            methionine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  AUTHORS   Yu JSL, Correia-Melo C, Zorrilla F, Herrera-Dominguez L, Wu MY, 
            Hartl J, Campbell K, Blasche S, Kreidl M, Egger AS, Messner CB, 
            Demichev V, Freiwald A, Mulleder M, Howell M, Berman J, Patil KR, 
            Alam MT, Ralser M
  TITLE     Microbial communities form rich extracellular metabolomes that 
            foster metabolic interactions and promote drug tolerance.
  JOURNAL   Nat Microbiol. 2022 Apr;7(4):542-555. doi: 
            10.1038/s41564-022-01072-5. Epub 2022 Mar 21.
  PUBMED    35314781
REFERENCE   2  (bases 1 to 6360)
  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"; doi: 
            "10.1038/s41564-022-01072-5"; journalName: "Nat Microbiol"; date: 
            "2022-04"; volume: "7"; issue: "4"; pages: "542-555"
COMMENT     Sequence Label: pM-TagRFP657
FEATURES             Location/Qualifiers
     source          1..6360
                     /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      1601..1788
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1656..1680)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1855..2358
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(2617..3951)
                     /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(4252..4271)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4253..4271)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4358..4375)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4529..5117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4609..4628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5288..6079,6080..6148))
                     /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     5911..5930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6149..6253)
                     /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 gagcgaattg atcacagaaa
      901 atatgcacat gaagttgtat atggagggaa ccgtcaataa ccatcatttc aaatgcactt
      961 ctgaaggtga gggcaaacct tacgaaggga cccaaacaca gagaattaag gtagtagaag
     1021 ggggtccact gccatttgcc tttgacatcc tagcaacatc cttcatgtac ggttcccaca
     1081 cattcataaa ccacactcaa ggaataccag acttttggaa gcagagtttt ccagaaggtt
     1141 ttacgtggga aagagttact acttacgagg acggtggtgt attaacagcc acacaagata
     1201 catcattaca agatggttgt ctgatctaca acgttaaaat cagaggtgtt aactttcctt
     1261 caaatgggcc agttatgcaa aagaaaactt taggctggga ggcccatact gaaatgcttt
     1321 acccagcaga tggcggcttg gaaggcagga cggctttggc tttaaagcta gttggtggag
     1381 gacatttgat ttgtaatttc aaaacaactt acagaagcaa aaagcctgct aaaaatctta
     1441 agatgccagg agtgtactat gtcgattaca gattggaacg tattaaagag gctgataagg
     1501 aaacctatgt ggaacaacat gaagtggcag tcgcgagata ttgtgatctt ccttctaaac
     1561 taggtcataa actcaattaa ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     1621 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1681 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     1741 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     1801 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt caccgatcgc
     1861 ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact
     1921 ctgtatttat ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag
     1981 aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
     2041 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
     2101 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
     2161 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
     2221 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     2281 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     2341 atttttatag cacgtgatga aaaggaccct aagaaaccat tattatcatg acattaacct
     2401 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2461 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2521 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg tcgagcttgt gagagaaagt
     2581 aggtttatac ataattttac aactcattac gcacactcat ggtttttggc cagcgaaaac
     2641 agtttcaaaa gattgctgga agtctgcaat aatgtcatca ataaattcga taccaacaga
     2701 gacacgaatt aagtccttgg taacaccaga tgccaacttt tctttgtcat ttaattgttt
     2761 gtgggtagtg aagtatggag caatgactaa ggtcttggca tcaccaacat tggccaagtt
     2821 agaggcaagc tttaaattgt caacaacttg agcaccagaa agtttgaatg ggtcagtttc
     2881 cttgtcggca tttggtaagt cttttacacc gaaagataag acaccaccga aaccgttaga
     2941 tagatacttc ttagcatttt catgatgaga atgagatgct aaaccagggt atgaaaccca
     3001 agatacgtat ggggattgtt ctaaccattt ggctaacttc gatgcatttt caccgtgtct
     3061 ttcagctctc aaagataatg tttcaacacc ttgtagtagc aagaaagagg caaatgggtt
     3121 catcaatgga cccaaatctc ttaatagttc agttctaaca tgaacgatgt atgccaagtt
     3181 accgtaggct tcattgtaga tagtaccgtg atatccttcg gcaggttgag agaattgagg
     3241 gaacttttct gggtagtcct tccatgggaa cttaccagag tcaacaataa taccaccgat
     3301 agtagtacca tgaccaccaa tccatttggt agcagaatgt gttacaatat cagcaccgta
     3361 tttaattggc tgacagaagt aaccaccggc accaaatgtg ttgtcaacga caactggaat
     3421 accgtgtttg tgagcaattg caacaatttt ttcaaaatcc ggaacattgt actttggatt
     3481 accaatggtt tccaaataaa cagccttggt tctttcatca aagacctttt cgaattcttc
     3541 tggattgtca ccttcaacaa atctagcctc gataccaaat cttttgaacg agattttgaa
     3601 ctggttataa gtaccaccgt ataagtaaga agtggaaacg atgttgtcac cagtgtgtgc
     3661 caaaccttgg atggcaaggg tttgagcggc ttgaccggag gaaacagcca aagcagcagc
     3721 accaccttct aaagcagcaa ttctttcttc caaaacatta ctggttgggt tttggaaacg
     3781 ggaatagacg taacctggaa cttctagacc aaacaattgc gaaccatgct tagagtttcc
     3841 gaaaacataa gaagtggtgg cgtaaattgg tacagctctg gatctgtgag cattgtcacc
     3901 agggttctct tggccggcgt gtagttgaac agtatcgaaa tgagatggca ttgtatggat
     3961 gggggtaata gaattgtatc tatgtatctg acgaccctgt attacacgaa ggaaatagta
     4021 gatgaaaaga caagagagca agaaaaagga aaaacttcct ttccaacaga cgctttactt
     4081 aaccttatat atatcttatt tttttcatcg agcgtgttca attggacaag gtgccacctt
     4141 ttcgacactt cggttattat gttacacagt tttcatgagg atggcggagc tcgaattcat
     4201 cgatgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt
     4261 gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg
     4321 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4381 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4441 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4501 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4561 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4621 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4681 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4741 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4801 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4861 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4921 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4981 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5041 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5101 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5161 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5221 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5281 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5341 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5401 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5461 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5521 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5581 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5641 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5701 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5761 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5821 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5881 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5941 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6001 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6061 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6121 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6181 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6241 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6301 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
//
