LOCUS       sequence-350292-        5786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mRuby2 for auxotrophic selection in the absence of 
            histidine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5786)
  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 5786)
  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: pH-mRuby2
FEATURES             Location/Qualifiers
     source          1..5786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        2905..3092
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3093..3755
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATLPNGTNDVPSTKGVLM"
     primer_bind     complement(3821..3840)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3822..3840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3927..3944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4098..4686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4178..4197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4857..5648,5649..5717))
                     /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     5480..5499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5718..5786,1..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 tttgtcggcg atccccctag agtcttttac
     2401 atcttcggaa aacaaaaact attttttctt taattccttt ttttactttc tatttttaat
     2461 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg
     2521 accctaagaa accattatta tcatgacatt aacctataaa aataggcgca tcacgaggcc
     2581 ctttcgtctc gcgtgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2641 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2701 agcgggtgtt ggcggacgtc ggggctggct taactatgcg gcatcagagc agattgtact
     2761 gagagtgcac cataaattcc cgttttaaga gcttggtgag cgctaggagt cactgccagg
     2821 tatcgtttga acacggcatt agtcagggaa gtcataacac agtcctttcc cgcaattttc
     2881 tttttctatt actcttggcc tcctctagta cactctatat ttttttatgc ctcggtaatg
     2941 attttcattt ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat
     3001 tatcacataa tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa
     3061 aaaatgagca ggcaagataa acgaaggcaa agatgacaga gcagaaagcc ctagtaaagc
     3121 gtattacaaa tgaaaccaag attcagattg cgatctcttt aaagggtggt cccctagcga
     3181 tagagcactc gatcttccca gaaaaggagg cagaagcagt agcagaacag gccacacaat
     3241 cgcaagtgat taacgtccac acaggtatag ggtttctgga ccatatgata catgctctgg
     3301 ccaagcattc cggctggtcg ctaatcgttg agtgcattgg tgacttacac atagacgacc
     3361 atcacaccac tgaagactgc gggattgctc tcggtcaagc ttttaaagag gccctagggg
     3421 ccgtgcgtgg agtaaaaagg tttggatcag gatttgcgcc tttggatgag gcactttcca
     3481 gagcggtggt agatctttcg aacaggccgt acgcagttgt cgaacttggt ttgcaaaggg
     3541 agaaagtagg agatctctct tgcgagatga tcccgcattt tcttgaaagc tttgcagagg
     3601 ctagcagaat taccctccac gttgattgtc tgcgaggcaa gaatgatcat caccgtagtg
     3661 agagtgcgtt caaggctctt gcggttgcca taagagaagc caccttgccc aatggtacca
     3721 acgatgttcc ctccaccaaa ggtgttctta tgtaggagct cgaattcatc gatgatatca
     3781 gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta
     3841 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5761 tgtctcatga gcggatacat atttga
//
