LOCUS       sequence-369377-        3057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes KanR 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 3057)
  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 3057)
  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: pMYT035
FEATURES             Location/Qualifiers
     source          1..3057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(614..1202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(694..713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1384..2043
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1450..1469)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2114..2148
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     gene            complement(join(2165..3057,1..463))
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(2165..2362)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(2367..3057,1..119))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
       61 tcagcatcca tgttggaatt taatcgcggc ctcgaaacgt gagtcttttc cttacccatt
      121 tttgtttatg ttcggatgtg atgtgagaac tgtatcctag caagatttta aaaggaagta
      181 tatgaaagaa gaacctcagt ggcaaatcct aaccttttat atttctctac aggggcgcgg
      241 cgtggggaca attcaacgcg tctgtgaggg gagcgtttcc ctgctcgcag gtctggagcg
      301 aggagccgta atttttgctt cgcgccgtgc ggccatcaaa atgtatggat gcaaatgatt
      361 atacatgggg atgtatgggc taaatgtacg ggcgacagtc acatcatgcc cctgagctgc
      421 gcacgtcaag actgtcaagg agggtattct gggcctccat gtcgctggcc gggtgacccg
      481 gcggggacga ggcaagctaa actgtaatgt cttcacggtt atccacagaa tcaggggata
      541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1261 aagggatttt ggtcatgatt gatcgggcac gtaagaggtt ccaactttca ccataatgaa
     1321 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     1381 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     1441 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     1501 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     1561 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
     1621 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     1681 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     1741 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     1801 gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     1861 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     1921 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     1981 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     2041 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact
     2101 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgaagacat
     2161 actccagtat agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac
     2221 ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat
     2281 ttgattaaaa tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa
     2341 caagaatctt tttattgtca gttactttag aaaaactcat cgagcatcaa atgaaactgc
     2401 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     2461 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     2521 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     2581 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt
     2641 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     2701 accaaaccgt tattcattcg tgattgcgcc tgagccagac gaaatacgcg atcgctgtta
     2761 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     2821 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttgccgggg
     2881 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     2941 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     3001 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaat
//
