LOCUS       sequence-369369-        3579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes LEU2 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 3579)
  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 3579)
  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: pMYT030
FEATURES             Location/Qualifiers
     source          1..3579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             178..1272
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     terminator      complement(1463..1497)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1568..2227)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2142..2161
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2409..2997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2898..2917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3349..3579,1..177)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
ORIGIN
        1 gaatagagaa gcgttcatga ctaaatgctt gcatcacaat acttgaagtt gacaatatta
       61 tttaaggacc tattgttttt tccaataggt ggttagcaat cgtcttactt tctaactttt
      121 cttacctttt acatttcagc aatatatata tatatatttc aaggatatac cattctaatg
      181 tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc
      241 gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa gttcgatttc
      301 gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgttccact tccagatgag
      361 gcgctggaag cctccaagaa ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa
      421 tggggaaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa agaacttcaa
      481 ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga cttatctcca
      541 atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt agtgggaggt
      601 atttactttg gtaagagaaa ggaggacgat ggtgatggtg tcgcttggga tagtgaacaa
      661 tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc cctacaacat
      721 gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc ttcaagatta
      781 tggagaaaaa ctgtggagga aaccatcaag aacgaatttc ctacattgaa ggttcaacat
      841 caattgattg attctgccgc catgatccta gttaagaacc caacccacct aaatggtatt
      901 ataatcacca gcaacatgtt tggtgatatc atctccgatg aagcctccgt tatcccaggt
      961 tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa caccgcattt
     1021 ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa ggtcaaccct
     1081 atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa
     1141 ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag aactggtgat
     1201 ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa
     1261 atccttgctt aacctggagg acccttctct ttagactatt ctactcttat gcacgtaaaa
     1321 aattctagga aatatgtatt aactaggagt aaaataaccg gctagtggca ttcatatagc
     1381 cgtctgttta catctacatc acacatttcg agtgtatatc tcgcaacgtt ggcgttaaat
     1441 aggcaggagt atgtcttcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     1501 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     1561 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     1621 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     1681 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     1741 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gaaacgaaaa
     1801 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     1861 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     1921 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     1981 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     2041 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     2101 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     2161 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     2221 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     2281 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aatcatgacc
     2341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2521 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3061 tttcctgcgt tatcccctga ttctgtggat aaccgtgaag acattacatc gaggagaact
     3121 tctagtatat ctacatacct aatattattg ccttattaaa aatggaatcc caacaattac
     3181 atcaaaatcc acattctctt caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc
     3241 aaaaacgtta tatttatagg ataattatac tctatttctc aacaagtaat tggttgtttg
     3301 gccgagcggt ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa ctgtgggaat
     3361 actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt tttttcctca
     3421 acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga ctggaaattt
     3481 tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa attgggagaa
     3541 aaaggaaagg tgagagcgcc ggaaccggct tttcatata
//
