LOCUS       sequence-110235-        3583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes LEU2 as a Type 6 part to be used in the Dueber YTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3583)
  AUTHORS   Lee ME, DeLoache WC, Cervantes B, Dueber JE
  TITLE     A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
  JOURNAL   ACS Synth Biol. 2015 Apr 14.
  PUBMED    25871405
REFERENCE   2  (bases 1 to 3583)
  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. 2015 Apr 14."
COMMENT     Sequence Label: pYTK075
FEATURES             Location/Qualifiers
     source          1..3583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        254..661
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             662..1756
                     /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(1948..1982)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2053..2712)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2627..2646
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2894..3482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3383..3402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcat acatcgagga gaacttctag tatatctaca tacctaatat tattgcctta
       61 ttaaaaatgg aatcccaaca attacatcaa aatccacatt ctcttcaaaa tcaattgtcc
      121 tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt ataggataat tatactctat
      181 ttctcaacaa gtaattggtt gtttggccga gcggtctaag gcgcctgatt caagaaatat
      241 cttgaccgca gttaactgtg ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc
      301 ttagcaacca ttattttttt cctcaacata acgagaacac acaggggcgc tatcgcacag
      361 aatcaaattc gatgactgga aattttttgt taatttcaga ggtcgcctga cgcatatacc
      421 tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac cggcttttca
      481 tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga agttgacaat
      541 attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac
      601 ttttcttacc ttttacattt cagcaatata tatatatata tttcaaggat ataccattct
      661 aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac
      721 agccgaagcc attaaggttc ttaaagctat ttctgatgtt cgttccaatg tcaagttcga
      781 tttcgaaaat catttaattg gtggtgctgc tatcgatgct acaggtgttc cacttccaga
      841 tgaggcgctg gaagcctcca agaaggctga tgccgttttg ttaggtgctg tgggtggtcc
      901 taaatgggga accggtagtg ttagacctga acaaggttta ctaaaaatcc gtaaagaact
      961 tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc gactctcttt tagacttatc
     1021 tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag aattagtggg
     1081 aggtatttac tttggtaaga gaaaggagga cgatggtgat ggtgtcgctt gggatagtga
     1141 acaatacacc gttccagaag tgcaaagaat cacaagaatg gccgctttca tggccctaca
     1201 acatgagcca ccattgccta tttggtcctt ggataaagct aatgttttgg cctcttcaag
     1261 attatggaga aaaactgtgg aggaaaccat caagaacgaa tttcctacat tgaaggttca
     1321 acatcaattg attgattctg ccgccatgat cctagttaag aacccaaccc acctaaatgg
     1381 tattataatc accagcaaca tgtttggtga tatcatctcc gatgaagcct ccgttatccc
     1441 aggttccttg ggtttgttgc catctgcgtc cttggcctct ttgccagaca agaacaccgc
     1501 atttggtttg tacgaaccat gccacggttc tgctccagat ttgccaaaga ataaggtcaa
     1561 ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa ttgtcattga acttgcctga
     1621 agaaggtaag gccattgaag atgcagttaa aaaggttttg gatgcaggta tcagaactgg
     1681 tgatttaggt ggttccaaca gtaccaccga agtcggtgat gctgtcgccg aagaagttaa
     1741 gaaaatcctt gcttaacctg gaggaccctt ctctttagac tattctactc ttatgcacgt
     1801 aaaaaattct aggaaatatg tattaactag gagtaaaata accggctagt ggcattcata
     1861 tagccgtctg tttacatcta catcacacat ttcgagtgta tatctcgcaa cgttggcgtt
     1921 aaataggcag gagttgagac cagaccaata aaaaacgccc ggcggcaacc gagcgttctg
     1981 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
     2041 ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     2101 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     2161 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     2221 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgaaac
     2281 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     2341 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     2401 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     2461 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg
     2521 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     2581 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     2641 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     2701 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     2761 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc cgatcaatca
     2821 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2881 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2941 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3001 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3061 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3541 tgttctttcc tgcgttatcc cctgattctg tggataaccg tag
//
