LOCUS       sequence-369375-        6208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes LYS2 as a yeast Vector selectable marker to be used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6208)
  AUTHORS   Shaw WM, Khalil AS, Ellis T
  TITLE     A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423.
  PUBMED    37930278
REFERENCE   2  (bases 1 to 6208)
  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"; journalName: "ACS Synth 
            Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423."
COMMENT     Sequence Label: pMYT034
FEATURES             Location/Qualifiers
     source          1..6208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(36..624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(116..135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             806..1465
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(872..891)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1536..1570
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1714..5892)
                     /codon_start=1
                     /gene="S. cerevisiae LYS2"
                     /product="alpha-aminoadipate reductase, required for lysine
                     biosynthesis"
                     /label=LYS2
                     /note="yeast auxotrophic marker"
                     /translation="MTNEKVWIEKLDNPTLSVLPHDFLRPQQEPYTKQATYSLQLPQLD
                     VPHDSFSNKYAVALSVWAALIYRVTGDDDIVLYIANNKILRFNIQPTWSFNELYSTINN
                     ELNKLNSIEANFSFDELAEKIQSCQDLERTPQLFRLAFLENQDFKLDEFKHHLVDFALN
                     LDTSNNAHVLNLIYNSLLYSNERVTIVADQFTQYLTAALSDPSNCITKISLITASSKDS
                     LPDPTKNLGWCDFVGCIHDIFQDNAEAFPERTCVVETPTLNSDKSRSFTYRDINRTSNI
                     VAHYLIKTGIKRGDVVMIYSSRGVDLMVCVMGVLKAGATFSVIDPAYPPARQTIYLGVA
                     KPRGLIVIRAAGQLDQLVEDYINDELEIVSRINSIAIQENGTIEGGKLDNGEDVLAPYD
                     HYKDTRTGVVVGPDSNPTLSFTSGSEGIPKGVLGRHFSLAYYFNWMSKRFNLTENDKFT
                     MLSGIAHDPIQRDMFTPLFLGAQLYVPTQDDIGTPGRLAEWMSKYGCTVTHLTPAMGQL
                     LTAQATTPFPKLHHAFFVGDILTKRDCLRLQTLAENCRIVNMYGTTETQRAVSYFEVKS
                     KNDDPNFLKKLKDVMPAGKGMLNVQLLVVNRNDRTQICGIGEIGEIYVRAGGLAEGYRG
                     LPELNKEKFVNNWFVEKDHWNYLDKDNGEPWRQFWLGPRDRLYRTGDLGRYLPNGDCEC
                     CGRADDQVKIRGFRIELGEIDTHISQHPLVRENITLVRKNADNEPTLITFMVPRFDKPD
                     DLSKFQSDVPKEVETDPIVKGLIGYHLLSKDIRTFLKKRLASYAMPSLIVVMDKLPLNP
                     NGKVDKPKLQFPTPKQLNLVAENTVSETDDSQFTNVEREVRDLWLSILPTKPASVSPDD
                     SFFDLGGHSILATKMIFTLKKKLQVDLPLGTIFKYPTIKAFAAEIDRIKSSGGSSQGEV
                     VENVTANYAEDAKKLVETLPSSYPSREYFVEPNSAEGKTTINVFVTGVTGFLGSYILAD
                     LLGRSPKNYSFKVFAHVRAKDEEAAFARLQKAGITYGTWNEKFASNIKVVLGDLSKSQF
                     GLSDEKWMDLANTVDIIIHNGALVHWVYPYAKLRDPNVISTINVMSLAAVGKPKFFDFV
                     SSTSTLDTEYYFNLSDKLVSEGKPGILESDDLMNSASGLTGGYGQSKWAAEYIIRRAGE
                     RGLRGCIVRPGYVTGASANGSSNTDDFLLRFLKGSVQLGKIPDIENSVNMVPVDHVARV
                     VVATSLNPPKENELAVAQVTGHPRILFKDYLYTLHDYGYDVEIESYSKWKKSLEASVID
                     RNEENALYPLLHMVLDNLPESTKAPELDDRNAVASLKKDTAWTGVDWSNGIGVTPEEVG
                     IYIAFLNKVGFLPPPTHNDKLPLPSIELTQAQISLVASGAGARGSSAAA"
     promoter        complement(5893..6132)
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
ORIGIN
        1 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
       61 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga ttgatcgggc acgtaagagg
      721 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
      781 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
      841 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
      901 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
      961 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1021 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1081 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1141 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1201 ctatttccct aaagggttta ttgagaatat gtttttcgtt tcagccaatc cctgggtgag
     1261 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1321 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1381 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1441 cgatgagtgg cagggcgggg cgtaatttga tatcgagctc gcttggactc ctgttgatag
     1501 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
     1561 cgttttttat tggtgaagac atactctatt ttattctctt attatatatt attctcggag
     1621 tttttaagtg acatcacccg aaaagaagct aagtctttct cctaattcat atttaattat
     1681 tgtacatgga catatcatac gtaatgctca accttaagct gctgcggagc ttccacgagc
     1741 accagcacct gaagcaacta gacttatttg cgcttgagtt agttctatac ttggcagtgg
     1801 aagtttgtca ttatgagttg gtggaggtaa aaatccaacc ttgtttaaaa atgcaatata
     1861 tataccaacc tcttctggag taacacctat tccattagac caatcaacac ctgtccatgc
     1921 ggtgtctttc tttaaagatg ccacggcgtt cctatcgtct agttccggag ctttggtact
     1981 ttcaggtaag ttgtctaaga ccatgtgtag caaaggatac aacgcatttt cttcattcct
     2041 gtcaataaca gacgcctcca atgatttctt ccatttagaa tagctttcga tttcgacatc
     2101 gtaaccataa tcgtgtaaag tatacaagta gtctttgaat aatattcttg ggtgacccgt
     2161 tacttgagca acggccaatt cattttcttt gggaggattc aaagacgtag caacaacaac
     2221 acgagcaaca tgatctactg gaaccatatt cacggaattt tcgatatctg gaatcttacc
     2281 taattggact gaacctttca aaaatctcaa taagaaatca tctgtgtttg aagaaccatt
     2341 ggcagaggca cctgttacgt aacctggtct gacaatacac ccacgtaggc ccctttcacc
     2401 tgcacgtcta atgatgtact cagcagccca tttggactga ccatatccac cagtgagccc
     2461 gcttgcagag ttcattaaat cgtctgattc taaaatgcct ggcttccctt cgctaacaag
     2521 tttatctgac aaattaaagt agtattcagt gtcaagagta gaagtggagg aaacaaagtc
     2581 aaagaacttt ggcttgccga cggcggctaa gctcataaca ttgatagttg aaataacatt
     2641 tggatctctc aatttggcat atggataaac ccagtgaact aacgcaccat tatggataat
     2701 tatatcaact gtgtttgcca aatccatcca cttctcatct gaaagaccaa attggctttt
     2761 agataaatcg cctaatacaa ctttaatatt tgaggcaaat ttttcgttcc aagtaccata
     2821 ggtgatacct gccttttgta atcttgcaaa tgcagcttct tcatccttgg ccctgacgtg
     2881 ggcaaacact ttgaaactgt agttctttgg agaacgtcct aacaaatctg caaggatgta
     2941 ggagcccaga aatcctgtga caccggtaac aaacacatta attgttgttt ttccttcggc
     3001 actattaggt tcaacaaaat attccctaga ggggtacgaa cttggtagcg tttcaaccaa
     3061 tttcttggca tcttccgcat aatttgcagt gacattttcg acgacctcac cttgagatga
     3121 tccacccgat gatttaattc tgtcaatttc cgcggcaaag gcctttatcg ttggatactt
     3181 gaaaattgtg cccaatggta aatcaacttg cagctttttc tttaaggtaa aaatcatttt
     3241 ggtagccaag atagaatgac cacctaaatc gaaaaacgaa tcatctggtg atacagatgc
     3301 tggcttggta ggtaatatac ttaaccataa gtctctaacc tcgcgctcaa cattggtaaa
     3361 ctgagagtcg tcagtttcag aaactgtatt ttcagctacc aaatttaatt gcttgggagt
     3421 tgggaattga agtttaggct tatcaacttt accatttgga ttcaatggta gtttatccat
     3481 aaccacaatc aaggaaggca tagcatagct agccaatctt ttctttaaga aagtcctgat
     3541 gtccttggat aaaagatggt aaccgattaa gcccttaact atagggtcag tttcaacctc
     3601 ctttggaaca tcactttgga acttagacaa gtcatctggc ttgtcaaatc ttgggaccat
     3661 aaatgtgatc aatgttggct cattgtcggc atttttgcga actaaagtaa tgttttctct
     3721 taccaatgga tgttgggaaa tgtgcgtatc tatttctcct aattcgattc tgaacccacg
     3781 aattttaact tgatcatcag ccctaccgca acattcacag tcaccgtttg gtagataacg
     3841 acctaaatca cccgttctgt acaatctatc tcttggacct aaccagaatt gtctccaagg
     3901 ttcaccatta tccttatcca aataattcca gtgatctttt tcaacaaacc agttgttcac
     3961 aaatttttct ttattcaatt ctggtaatcc tctataacct tcggccaaac cacctgcacg
     4021 aacataaatc tcacctattt cgccaatacc acatatttga gtacgatcgt tcctgttaac
     4081 aactaatagc tgaacgttca acataccttt accagcaggc atgacatctt tcaatttttt
     4141 caaaaagttt ggatcgtcat tttttgattt aacttcgaaa taagaaactg cacgctgtgt
     4201 ttcagtggta ccgtacatat taacaatacg gcaattttct gccaaggttt gtaacctcag
     4261 acaatcacgt tttgttaaaa tgtcacccac aaagaacgca tgatgtaact tagggaatgg
     4321 tgtagtagct tgggcagtaa gtaattgacc catggcaggt gttaaatggg taactgtgca
     4381 accatactta ctcatccatt ccgctaaacg gcccggtgta ccaatatcat cttgagtagg
     4441 gacatacaat tgggcaccta aaaataatgg tgtaaacata tctctttgaa ttggatcatg
     4501 tgcaataccg ctcagcattg tgaatttatc attttctgtt aagttgaacc ttttggacat
     4561 ccaattgaaa taataagcca aggaaaaatg tctaccaaga acacccttag gaataccttc
     4621 ggaaccagat gtgaaagata gggttgggtt ggaatctggt ccaactacaa cacctgttct
     4681 ggtgtctttg tagtgatcat atggagccaa aacatcctcg ccattgtcca atttgccacc
     4741 ttcaatggta ccattttctt gaatagcgat ggaattgatt cttgaaacaa tctccaattc
     4801 atcattgatg taatcttcta ccaattgatc caattgtcca gcagctctaa taacaatcaa
     4861 cccacgtggt ttagcaacac ctaagtaaat ggtttgtctg gctgggggat atgcagggtc
     4921 gataactgaa aaggttgcgc cggctttcaa gacacccatc acacatacca tcaaatccac
     4981 acccctagaa gaatagatca tcactacatc acctcttttg atacctgttt taatcaaata
     5041 atgggcaact atgttagaag tgcggttgat gtcgcgataa gtgaaagaac gggacttgtc
     5101 ggaatttagt gttggagtct ccacaacaca ggttctctct gggaaggctt cagcattgtc
     5161 ctggaaaatg tcgtgaatac accccacgaa atcgcaccag cccaagttct tagttggatc
     5221 aggtaaacta tccttggatg atgcggtgat cagagagatt ttagttatgc aattggatgg
     5281 atcgcttagc gcagcagtca aatattgagt aaattggtcc gcaacaatgg ttactctttc
     5341 attcgaatac agtaagctgt tataaattaa gttcaaaaca tgcgcattat tactggtatc
     5401 caaattcaaa gcaaagtcca ctaaatgatg cttgaactcg tctaatttga aatcttggtt
     5461 ttccaaaaag gccaaacgga acaactgagg ggtcctttcc aaatcttggc aactttgaat
     5521 tttttcagct agctcgtcaa aggaaaaatt ggcctcaata gaattgagct tgttcaactc
     5581 attgttaatt gtagaataca gctcattaaa tgaccacgtt ggttgaatat tgaatcttaa
     5641 gattttgtta ttcgcaatat aaagaacaat atcatcgtca ccggttactc tatatatcaa
     5701 tgcagcccat acactcaaag cgacagcgta tttgttagaa aaactatcat gaggcacatc
     5761 gagctgaggt agctgtaacg aatatgtagc ttgtttcgta taaggttctt gttgtgggcg
     5821 taaaaagtca tgtggtaaca ctgaaagagt tggattatcc aacttctcta tccagacctt
     5881 ttcgttagtc attagtgagt aactctgtga tatctctcta taattagcag tttttcactg
     5941 aaattccagg aaaggtaata aactcagatt tttttttata ctattggctg cttgttactt
     6001 atatatcttg aacttctccc agcgggtctt gaaatacatt tgggcgatgt tcatgttcat
     6061 taggcaggta tttcgacatt gagtcacacg cgaaaaaccg ccggaatttt ttatgtaatt
     6121 gcaagtggaa tttgtaatgt cttcacggtt atccacagaa tcaggggata acgcaggaaa
     6181 gaacatgtga gcaaaaggcc agcaaaag
//
