LOCUS       sequence-350989-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mRuby2 for auxotrophic selection in the absence of 
            methionine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  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 6372)
  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-mRuby2
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     282..301
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        320..986
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     856..879
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1022..1732
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     terminator      1756..1943
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1811..1835)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    2010..2513
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(2772..4106)
                     /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(4407..4426)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4408..4426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4513..4530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4684..5272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4764..4783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5443..6234,6235..6303))
                     /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     6066..6085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6304..6372,1..36))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
       61 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      121 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      181 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      241 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      301 actgagagtg caccatatgt cattatcaat actgccattt caaagaatac gtaaataatt
      361 aatagtagtg attttcctaa ctttatttag tcaaaaaatt agccttttaa ttctgctgta
      421 acccgtacat gcccaaaata gggggcgggt tacacagaat atataacatc gtaggtgtct
      481 gggtgaacag tttattcctg gcatccacta aatataatgg agcccgcttt ttaagctggc
      541 atccagaaaa aaaaagaatc ccagcaccaa aatattgttt tcttcaccaa ccatcagttc
      601 ataggtccat tctcttagcg caactacaga gaacaggggc acaaacaggc aaaaaacggg
      661 cacaacctca atggagtgat gcaacctgcc tggagtaaat gatgacacaa ggcaattgac
      721 ccacgcatgt atctatctca ttttcttaca ccttctatta ccttctgctc tctctgattt
      781 ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa attattcccc tacttgacta
      841 ataagtatat aaagacggta ggtattgatt gtaattctgt aaatctattt cttaaacttc
      901 ttaaattcta cttttatagt tagtcttttt tttagtttta aaacaccaag aacttagttt
      961 cgaataaaca cacataaaca aacaaagtcg acggatcggt gacggtgctg gtttaattaa
     1021 catggtgtcc aaaggagagg agttaatcaa ggaaaacatg agaatgaaag ttgtcatgga
     1081 gggctccgtt aatggtcacc aattcaagtg tacaggggaa ggtgaaggta atccttacat
     1141 gggtacacaa actatgagaa ttaaagtaat tgaaggcgga ccactaccat ttgcatttga
     1201 cattctggca acgtcattca tgtacggatc acgaactttc atcaagtacc ctaaaggtat
     1261 accagacttt ttcaagcaat cttttccaga gggttttaca tgggaaaggg ttacaagata
     1321 cgaagatggg ggtgtcgtca cagttatgca agatacttca ttagaagatg gctgccttgt
     1381 ctatcatgtg caagtaagag gggtgaattt tccttctaac ggacctgtga tgcagaaaaa
     1441 gaccaaaggt tgggaaccaa atactgaaat gatgtaccca gctgatggag gtttgagagg
     1501 ctacacacac atggcgctta aagttgatgg tggaggtcat ttgtcttgta gttttgttac
     1561 cacttatcgt tctaaaaaga ctgttggcaa tatcaaaatg ccaggaatac atgctgtaga
     1621 ccacagacta gaaagactcg aagagagcga taacgaaatg ttcgttgtac agagagagca
     1681 tgccgtagcc aaatttgctg gcttaggcgg tggtatggat gaattgtata agtaaggcgc
     1741 gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
     1801 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     1861 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct
     1921 cttattgacc acacctctac cggcagatcc gctagggata acagggtaat atagatctgt
     1981 ttagcttgcc tcgtccccgc cgggtcaccg atcgcttgcc tgtaacttac acgcgcctcg
     2041 tatcttttaa tgatggaata atttgggaat ttactctgta tttatttatt tttatgtttt
     2101 gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa
     2161 aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga
     2221 caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac
     2281 aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg
     2341 agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac
     2401 atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat
     2461 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg
     2521 accctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     2581 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2641 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2701 agcgggtgtt ggcggtcgag cttgtgagag aaagtaggtt tatacataat tttacaactc
     2761 attacgcaca ctcatggttt ttggccagcg aaaacagttt caaaagattg ctggaagtct
     2821 gcaataatgt catcaataaa ttcgatacca acagagacac gaattaagtc cttggtaaca
     2881 ccagatgcca acttttcttt gtcatttaat tgtttgtggg tagtgaagta tggagcaatg
     2941 actaaggtct tggcatcacc aacattggcc aagttagagg caagctttaa attgtcaaca
     3001 acttgagcac cagaaagttt gaatgggtca gtttccttgt cggcatttgg taagtctttt
     3061 acaccgaaag ataagacacc accgaaaccg ttagatagat acttcttagc attttcatga
     3121 tgagaatgag atgctaaacc agggtatgaa acccaagata cgtatgggga ttgttctaac
     3181 catttggcta acttcgatgc attttcaccg tgtctttcag ctctcaaaga taatgtttca
     3241 acaccttgta gtagcaagaa agaggcaaat gggttcatca atggacccaa atctcttaat
     3301 agttcagttc taacatgaac gatgtatgcc aagttaccgt aggcttcatt gtagatagta
     3361 ccgtgatatc cttcggcagg ttgagagaat tgagggaact tttctgggta gtccttccat
     3421 gggaacttac cagagtcaac aataatacca ccgatagtag taccatgacc accaatccat
     3481 ttggtagcag aatgtgttac aatatcagca ccgtatttaa ttggctgaca gaagtaacca
     3541 ccggcaccaa atgtgttgtc aacgacaact ggaataccgt gtttgtgagc aattgcaaca
     3601 attttttcaa aatccggaac attgtacttt ggattaccaa tggtttccaa ataaacagcc
     3661 ttggttcttt catcaaagac cttttcgaat tcttctggat tgtcaccttc aacaaatcta
     3721 gcctcgatac caaatctttt gaacgagatt ttgaactggt tataagtacc accgtataag
     3781 taagaagtgg aaacgatgtt gtcaccagtg tgtgccaaac cttggatggc aagggtttga
     3841 gcggcttgac cggaggaaac agccaaagca gcagcaccac cttctaaagc agcaattctt
     3901 tcttccaaaa cattactggt tgggttttgg aaacgggaat agacgtaacc tggaacttct
     3961 agaccaaaca attgcgaacc atgcttagag tttccgaaaa cataagaagt ggtggcgtaa
     4021 attggtacag ctctggatct gtgagcattg tcaccagggt tctcttggcc ggcgtgtagt
     4081 tgaacagtat cgaaatgaga tggcattgta tggatggggg taatagaatt gtatctatgt
     4141 atctgacgac cctgtattac acgaaggaaa tagtagatga aaagacaaga gagcaagaaa
     4201 aaggaaaaac ttcctttcca acagacgctt tacttaacct tatatatatc ttattttttt
     4261 catcgagcgt gttcaattgg acaaggtgcc accttttcga cacttcggtt attatgttac
     4321 acagttttca tgaggatggc ggagctcgaa ttcatcgatg atatcagatc cactagtggc
     4381 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
     4441 ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4501 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4561 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4621 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4681 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4741 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4801 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4861 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4921 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4981 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5041 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5101 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5161 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5221 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6361 atacatattt ga
//
