LOCUS       sequence-474389-        4035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minimal centromeric shuttle vector depleted of restriction sites 
            outside the polylinker region, yeast selection marker LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4035)
  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 4035)
  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: pLS415
FEATURES             Location/Qualifiers
     source          1..4035
                     /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..2131)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2114..2130)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2131..2535
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2536..3630
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
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 aactgtggga atactcaggt atcgtaagat
     2161 gcaagagttc gagtctctta gcaaccatta tttttttcct caacataacg agaacacaca
     2221 ggggcgctat cgcacagaat caaattcgat gactggaaat tttttgttaa tttcagaggt
     2281 cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa ggtgagaggc
     2341 cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat
     2401 acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat
     2461 cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata tatatttcaa
     2521 ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg ggaccacgtt
     2581 ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga tgttcgttcc
     2641 aatgtcaagt tcgatttcga gaatcattta attggtggtg ctgctatcga cgctactggt
     2701 gtcccacttc cagatgaggc tctggaagcc tccaagaagg ctgatgccgt tttgttaggt
     2761 gctgtgggtg gtcctaaatg gggtacgggt agtgttagac cagaacaagg tttactaaaa
     2821 atccgtaaag aacttcaatt atacgccaat ttaagaccat gtaactttgc atccgattct
     2881 cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt cgttgttgtc
     2941 agagaattag tgggaggtat ttactttggt aagagaaagg aggacgatgg tgatggtgtc
     3001 gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag aatggcggct
     3061 ttcatggccc tacaacatga gccaccttta cctatttggt ccttagataa agctaatctt
     3121 ttggcctcat caagattatg gagaaaaact gtggaggaaa ccatcaagaa cgagtttcct
     3181 acattgaagg ttcaacatca gttgattgat tctgccgcca tgatcctagt taagaaccca
     3241 acccacctaa acggtattat tatcaccagc aatatgtttg gtgatattat ctccgatgaa
     3301 gcctccgtta tcccaggttc gttgggtttg ttgccatctg cgtcgttggc ctctttgcca
     3361 gacaagaaca ccgcatttgg tttgtacgaa ccatgtcacg gttctgcgcc agatttgcca
     3421 aagaataagg ttgatcctat cgccactatc ttgtctgcgg ctatgatgtt gaaattgtca
     3481 ttgaacttgc cagaagaagg taaggccatt gaagatgcag ttaaaaaggt tttggatgca
     3541 ggaatcagaa ctggtgattt aggtggttcc aatagtacca ccgaagtcgg tgatgctgtc
     3601 gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg atatttatac
     3661 ataaactata taaatgaaat tcataataga aacgacacga aattacaaaa tggaatatgt
     3721 tcataggata tacgaaacta tatacgcaat ctacatacat ttatcaagaa ggagaaaaag
     3781 gaggatagta aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa
     3841 aaagaattgc actttaacat taaaattgac aaggaatagg tcaccacaca aaaagttagg
     3901 tgtaacagaa aatcataaaa ctacgattcc taatttgata ttggaggatt ttctctaaaa
     3961 aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg gagtttaagt
     4021 caataccttc ttgaa
//
