LOCUS       sequence-353605-        5774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TagRFP657 for auxotrophic selection in the absence of 
            lysine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5774)
  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 5774)
  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-TagRFP657
FEATURES             Location/Qualifiers
     source          1..5774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        2750..2937
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2938..3600
                     /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(3666..3685)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3667..3685)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3772..3789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3943..4531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4023..4042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4702..5493,5494..5562))
                     /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     5325..5344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5563..5667)
                     /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 cggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     2281 ccttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     2341 atttttatag cacgtgatga aaaggaccct aagaaaccat tattatcatg acattaacct
     2401 ataaaaatag gcgcatcacg aggccctttc gtctcgcgtg tttcggtgat gacggtgaaa
     2461 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2521 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg acgtcggggc tggcttaact
     2581 atgcggcatc agagcagatt gtactgagag tgcaccataa attcccgttt taagagcttg
     2641 gtgagcgcta ggagtcactg ccaggtatcg tttgaacacg gcattagtca gggaagtcat
     2701 aacacagtcc tttcccgcaa ttttcttttt ctattactct tggcctcctc tagtacactc
     2761 tatatttttt tatgcctcgg taatgatttt catttttttt tttccaccta gcggatgact
     2821 cttttttttt cttagcgatt ggcattatca cataatgaat tatacattat ataaagtaat
     2881 gtgatttctt cgaagaatat actaaaaaat gagcaggcaa gataaacgaa ggcaaagatg
     2941 acagagcaga aagccctagt aaagcgtatt acaaatgaaa ccaagattca gattgcgatc
     3001 tctttaaagg gtggtcccct agcgatagag cactcgatct tcccagaaaa ggaggcagaa
     3061 gcagtagcag aacaggccac acaatcgcaa gtgattaacg tccacacagg tatagggttt
     3121 ctggaccata tgatacatgc tctggccaag cattccggct ggtcgctaat cgttgagtgc
     3181 attggtgact tacacataga cgaccatcac accactgaag actgcgggat tgctctcggt
     3241 caagctttta aagaggccct aggggccgtg cgtggagtaa aaaggtttgg atcaggattt
     3301 gcgcctttgg atgaggcact ttccagagcg gtggtagatc tttcgaacag gccgtacgca
     3361 gttgtcgaac ttggtttgca aagggagaaa gtaggagatc tctcttgcga gatgatcccg
     3421 cattttcttg aaagctttgc agaggctagc agaattaccc tccacgttga ttgtctgcga
     3481 ggcaagaatg atcatcaccg tagtgagagt gcgttcaagg ctcttgcggt tgccataaga
     3541 gaagccacct tgcccaatgg taccaacgat gttccctcca ccaaaggtgt tcttatgtag
     3601 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     3661 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     3721 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4381 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4561 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4681 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4741 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4801 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4861 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4921 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4981 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5041 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5101 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5161 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5221 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5281 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5341 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5401 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5461 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5521 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5581 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5641 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5701 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5761 tctcgcgcgt ttcg
//
