LOCUS       sequence-467224-        2360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid for MoClo, encodes inducible PGK1-tetO2p for K. 
            marxianus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2360)
  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 2360)
  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: pMTKM32
FEATURES             Location/Qualifiers
     source          1..2360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1272..1290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1293..1311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      complement(1415..1449)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1520..2179)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2094..2113
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtag tcggtctcaa acgttaccct cactctttca
      721 cattaccctc cccaccatat cttccccccc ccagcttcct gtcttccact atccctgcaa
      781 ccaccaccac cacacaacaa acatccccca gcagcccgga aaaatccacg cctccccccg
      841 aacacacgaa gcccgccccg gcttccactc caacactgcc aacactggaa ccccgcacca
      901 cgtaaccacc cgctttagtg gaaccccgcc cctcctgcgc ttcccttgcg cttcccttag
      961 gcttccctgg ggcttccctg gggcttccct tgcgaccaca ctccggtggc atggccctcg
     1021 cttgctacat gctacctttt ttttgccctt ttctggcctt ttctggcttt tctggcctca
     1081 ccacagtgta gtggtgtacc acagtgtaat ggtgtaacat tgtcgaatgg tattgtagcc
     1141 tgttgtagcc tgttgtagcc tgttgtagcc tggtgtaatg tctggcatag tctggtgtct
     1201 ggtgtctggt gtcttttgga aactgaacag tggaaagtgg aagtgaaaaa ttgtataaat
     1261 ataggtgtcc ctccctatca gtgatagaga tatccctatc agtgatagag aagacttgat
     1321 aagtatttcc ctagtaatac cccaccaacc atcggttgat acattttaat acacactacc
     1381 tatataaaga tacaaaatat gtgagaccag accaataaaa aacgcccggc ggcaaccgag
     1441 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     1501 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     1561 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1621 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     1681 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     1741 ctgaaacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     1801 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     1861 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     1921 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     1981 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     2041 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2101 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2161 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2221 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga
     2281 tcaatcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2341 gaaaagatca aaggatcttc
//
