LOCUS       sequence-467225-        2326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid for MoClo, encodes inducible TEF3-tetO2p for K. 
            marxianus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2326)
  AUTHORS   Thornbury M, Knoops A, Summerby-Murray I, Dhaliwal J, Johnson S, 
            Utomo JC, Joshi J, Narcross L, Remondetto G, Pouliot M, Whiteway M, 
            Martin VJJ
  TITLE     Sequencing of a Dairy Isolate Unlocks Kluyveromyces marxianus as a 
            Host for Lactose Valorization.
  JOURNAL   ACS Synth Biol. 2025 Jul 8. doi: 10.1021/acssynbio.5c00096.
  PUBMED    40629255
REFERENCE   2  (bases 1 to 2326)
  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. 2025 Jul 8. doi: 10.1021/acssynbio.5c00096."
COMMENT     Sequence Label: pMTKM31
FEATURES             Location/Qualifiers
     source          1..2326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(7..41)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(112..771)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     686..705
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      953..1541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1442..1461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2213..2231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2234..2252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 agaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
       61 gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc
      121 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
      181 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
      241 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
      301 atcaaaactg gtgaaactca cccagggatt ggctgaaacg aaaaacatat tctcaataaa
      361 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
      421 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
      481 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
      541 cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
      601 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
      661 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
      721 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
      781 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
      841 atggtgaaag ttggaacctc ttacgtgccc gatcaatcat gaccaaaatc ccttaacgtg
      901 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      961 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1021 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1081 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1141 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1201 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1261 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1321 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1381 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1441 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1501 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1561 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1621 ctgattctgt ggataaccgt agtcggtctc aaacgaagca aagaagtaac agcaggaaag
     1681 aaaaacaaac acaacaaaaa aaaacaagca gcatagcatc aacagaaatt tctaaagaga
     1741 accaaattca ccccagaaac aaccgcacaa atacgacatc catccacctt tcttttatct
     1801 tctttttctg atctgataat tagtttcata tacaatacgt agaaacaggc gcacagcacc
     1861 cagacctggc ttctgcccca gtgtataagc aatgtagcat aattggaaaa aaaaacgaaa
     1921 aataccgaaa ataagtggga agctgggcca caggagtggg gcgggatgcg actggttctg
     1981 agcgggaccg ggtaataagg ttgaaaaact ttgaattgat ggaataagaa acttctttct
     2041 tttcgctggc gggagaagga aaaaaaaatt ttttttttcc ttctgtttag tactggaaca
     2101 ttgagaaggc gtgtcaattt tgaataatta gagtggtcaa aaaaattttt tttgcttggg
     2161 ataccctttt tcgataatgt aaattttttt tgaatataaa aggagattga aatccctatc
     2221 agtgatagag atatccctat cagtgataga gattgatttg tagtttattt actagtattc
     2281 tcatagttct aactccaaga gaagtaacat taaagtatgt gagacc
//
