LOCUS       sequence-474392-        3487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minimal centromeric shuttle vector depleted of restriction sites 
            outside the polylinker region, yeast selection marker KanMX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3487)
  AUTHORS   Scutteri L, Barth P, Rahi SJ
  TITLE     Minimal shuttle vectors for Saccharomyces cerevisiae.
  JOURNAL   Synth Biol (Oxf). 2025 May 21;10(1):ysaf010. doi: 
            10.1093/synbio/ysaf010. eCollection 2025.
  PUBMED    40612073
REFERENCE   2  (bases 1 to 3487)
  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: "Synth Biol 
            (Oxf)."; date: "2025-05-21"; volume: "10(1):ysaf010. doi"; pages: " 
            10.1093/synbio/ysaf010. eCollection 2025"
COMMENT     Sequence Label: pLS410
FEATURES             Location/Qualifiers
     source          1..3487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..64
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(283..302)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      948..1536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1437..1456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1537..2040
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2041..2057
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2041..2057
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2114..2130)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            2131..3487
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2131..2474
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2475..3284
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      3290..3487
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
       61 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      121 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgcagaag atcagttggg
      181 tgcgcgagtg ggttacatcg aactggacct caacagtggt aagatactag agagttttcg
      241 ccccgaagaa cgattcccaa tgatgtccac ttttaaggtt ctgctatgtg gcgcggtatt
      301 atcccgtatt gacgcggggc aagagcaact aggtcgccgc atacactatt ctcagaatga
      361 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      421 attatgtagt gctgccataa cgatgagtga taacacggcg gcgaacttac ttctgacaac
      481 gattggagga ccaaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      541 ccttgatcgt tgggaaccag agctgaatga agccatacca aacgacgagc gtgacaccac
      601 gatgcctgtt gctatggcaa caaccttgcg gaaactatta actggcgaac tacttactct
      661 agcttcccgg caacagttaa tagactggat ggaggctgat aaagttgcag gaccacttct
      721 gcgctctgcg cttccggctg gctggtttat tgctgataaa tctggggccg gagagcgtgg
      781 gtcacgcggt atcatagcag cactagggcc agatggtaag ccctcccgta tcgtagttat
      841 ctacacgacg gggagccagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      901 tgcgtcactg attaagcatt ggtagtagaa aagatcaaag gatcttcttg agatcctttt
      961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1081 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1501 ttgtgatgct cgtcaggggg gcggagccta tggaaaatca cgtgctataa aaataattat
     1561 aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa
     1621 agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat
     1681 actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt
     1741 ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga
     1801 atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt
     1861 tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct
     1921 tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa
     1981 ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc
     2041 caggaaacag ctatgacgcg cgcgagctcg gtacccggga tcctctagag tcgacctgca
     2101 ggcatgccct aggactggcc gtcgttttac gacatggagg cccagaatac cctccttgac
     2161 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     2221 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     2281 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     2341 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     2401 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     2461 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaactcca
     2521 atatggatgc tgatttatat gggtataaat gggctaggga taatgtcggg caatcaggtg
     2581 cgacaatcta ccgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2641 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggagt
     2701 ttatgcctct accgaccatc aagcatttta tccgtactcc tgatgacgca tggttgctca
     2761 ccacagcgat ccccggcaaa acagctttcc aggtattaga agaatatcca gattcaggtg
     2821 aaaacattgt tgatgcgctg gctgtgttcc tgcgtcggtt gcactcgatt cctgtttgta
     2881 attgtccttt taacagtgat cgcgtatttc gactcgcaca ggcgcaatca cgaatgaata
     2941 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     3001 tctggaaaga aatgcacaaa cttttgccat tctcaccaga ttcagtcgtc actcatggtg
     3061 atttctcact tgataacctt atttttgacg aggggaaatt gataggttgt attgatgtag
     3121 gacgagtagg aatcgcagac cgctaccaag accttgcaat cctatggaac tgcctcggtg
     3181 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     3241 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcaata ctgacaataa
     3301 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     3361 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     3421 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgatggtcgc
     3481 tatactg
//
