LOCUS       sequence-459791-        2983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minimal integrating 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 2983)
  AUTHORS   Scutteri L, Barth P, Rahi SJ
  TITLE     Minimal integrating shuttle vectors for Saccharomyces cerevisiae 
            depleted of restriction sites outside the polylinker region
  JOURNAL   bioRxiv 2024.11.05.622133
REFERENCE   2  (bases 1 to 2983)
  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: "bioRxiv 
            2024.11.05.622133"
COMMENT     Sequence Label: pLS400
FEATURES             Location/Qualifiers
     source          1..2983
                     /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"
     primer_bind     1537..1553
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1537..1553
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1610..1626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            1627..2983
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1627..1970
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1971..2780
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2786..2983
                     /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 tggaaacagg aaacagctat gacgcgcgcg
     1561 agctcggtac ccgggatcct ctagagtcga cctgcaggca tgccctagga ctggccgtcg
     1621 ttttacgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     1681 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     1741 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
     1801 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
     1861 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
     1921 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg
     1981 aaaagactca cgtttcgagg ccgcgattaa actccaatat ggatgctgat ttatatgggt
     2041 ataaatgggc tagggataat gtcgggcaat caggtgcgac aatctaccga ttgtatggga
     2101 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2161 cagatgagat ggtcagacta aactggctga cggagtttat gcctctaccg accatcaagc
     2221 attttatccg tactcctgat gacgcatggt tgctcaccac agcgatcccc ggcaaaacag
     2281 ctttccaggt attagaagaa tatccagatt caggtgaaaa cattgttgat gcgctggctg
     2341 tgttcctgcg tcggttgcac tcgattcctg tttgtaattg tccttttaac agtgatcgcg
     2401 tatttcgact cgcacaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2461 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cacaaacttt
     2521 tgccattctc accagattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2581 ttgacgaggg gaaattgata ggttgtattg atgtaggacg agtaggaatc gcagaccgct
     2641 accaagacct tgcaatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2701 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2761 tgctcgatga gtttttctaa tcaatactga caataaaaag attcttgttt tcaagaactt
     2821 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     2881 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     2941 aatatcatgc gtcaatcgta tgtgaatgat ggtcgctata ctg
//
