LOCUS       sequence-464920-        2375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid for MoClo, encodes inducible TDH3-tetO2p for K. 
            marxianus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2375)
  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 2375)
  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: pMTKM30
FEATURES             Location/Qualifiers
     source          1..2375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     493..512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1286..1304
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1307..1325
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      complement(1433..1467)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1538..2197)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2112..2131
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
       61 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      661 tgcgttatcc cctgattctg tggataaccg tagtcggtct caaacgcact atatcaggcc
      721 tccactattc cacgcttcca ctatttcagg tttttccgac cacttattcc gatccggcag
      781 ggttcatcct ctcacctccc cccgctgcgc acccgtctaa ccaccctcgg catgacgaat
      841 tcactcgaga ggccctcatc cacacaacat atggctaaag tggtgaaact tgggccatga
      901 caaaggccat ggaaaatggt agtaccatgg tactaccagt ggtactacca ctagtggtgg
      961 cattagtggt accagaacca cctgttgatt ggtgctggga ctcctgtatt ttggttaggg
     1021 cctgtctgtt ggggcctgtc aaacaaccaa caacactctt ccatattctg ttctatccag
     1081 ctagttagtt agctagctag ctagctagct agctgtcttt ctctctcgta tgacatggcc
     1141 taaggccatg tcatgtcaca catacacata cacaccggtt taccgtttca gtaaatatcc
     1201 aacatgcgat catgccaata tcctgccaaa tattctttct accattattc tttctgccat
     1261 tctgccatat ataaagacca cattttccct atcagtgata gagatatccc tatcagtgat
     1321 agagaagtct ataataatta tatctcttcc ttgttcctcc caaaccaagt cttttagatt
     1381 taacaacagt acacacacac ttttacacat tcacatatgt gagaccagac caataaaaaa
     1441 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     1501 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     1561 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1621 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1681 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1741 aactcaccca gggattggct gaaacgaaaa acatattctc aataaaccct ttagggaaat
     1801 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1861 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1921 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     1981 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     2041 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     2101 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     2161 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     2221 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     2281 aacctcttac gtgcccgatc aatcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2341 tgagcgtcag accccgtaga aaagatcaaa ggatc
//
