LOCUS       sequence-474388-        4352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minimal integrating 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 4352)
  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 4352)
  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: pLS405_YCR051W
FEATURES             Location/Qualifiers
     source          1..4352
                     /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(2431..2448)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2431..2447)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2448..2852
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2853..3947
                     /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 tggaaacagg aaacagctat gacgcgcgct
     1561 aatgtgttgg acaacgacgg cgataccccg ttgcaccatg tggaggatgt ggccactgcc
     1621 aggttgatcg tggaagagct gggtggagac ttcactatca ggaatgtgga gggccaaacg
     1681 ccatacgact cgttcgtcga gaacggtgaa gatggtgagc taatcgagta catgaggatt
     1741 aagtccggcg tggccgatgt tcacggagtg gacggcgtgc agggtgaggg tgtcatcgac
     1801 agcaaattgc tggaagagtt caaggacaac gtgagataca ccttggaaaa tgaccctgag
     1861 gaaggagccg atgaggccac tctgcaacgc aggaggcagt tggaacagat cattacggga
     1921 gacaacgctg aggaggagtt ggaaaggtac atccgtgcta tggtcagaga gcgctccacc
     1981 gtggtcaaag acaggggcaa agagctccta ggtctatata tatatctata tacatattta
     2041 tatatattat tagaacttta caatatagta tataccattc attgtttaag tttcgggtaa
     2101 tacttttttt ttccttgtca taaccccaaa aattttcgat gcctttgata taattgagaa
     2161 caagaagagt ttgcaggtga caaaaatcga tgattatagg tgttgtgacg acaaaatgaa
     2221 cgctaatata tgggtggctg cttcagatgg taatttggac cgagtggaac atatcctccg
     2281 cgagagtaaa ggcgccatga ccccgcaatc caaggacatt aacggctaca ctccaatgca
     2341 tgctgccgcc gcatacggcc acctggattt gctgaagaaa atgtgcaatg agtacaatgg
     2401 agacatgtcg acctgcaggc atgccctagg actggccgtc gttttacaac tgtgggaata
     2461 ctcaggtatc gtaagatgca agagttcgag tctcttagca accattattt ttttcctcaa
     2521 cataacgaga acacacaggg gcgctatcgc acagaatcaa attcgatgac tggaaatttt
     2581 ttgttaattt cagaggtcgc ctgacgcata tacctttttc aactgaaaaa ttgggagaaa
     2641 aaggaaaggt gagaggccgg aaccggcttt tcatatagaa tagagaagcg ttcatgacta
     2701 aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat tgttttttcc
     2761 aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat
     2821 atatatatat atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt
     2881 tgccagggga ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta
     2941 tttctgatgt tcgttccaat gtcaagttcg atttcgagaa tcatttaatt ggtggtgctg
     3001 ctatcgacgc tactggtgtc ccacttccag atgaggctct ggaagcctcc aagaaggctg
     3061 atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg tacgggtagt gttagaccag
     3121 aacaaggttt actaaaaatc cgtaaagaac ttcaattata cgccaattta agaccatgta
     3181 actttgcatc cgattctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta
     3241 ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggagg
     3301 acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa
     3361 tcacaagaat ggcggctttc atggccctac aacatgagcc acctttacct atttggtcct
     3421 tagataaagc taatcttttg gcctcatcaa gattatggag aaaaactgtg gaggaaacca
     3481 tcaagaacga gtttcctaca ttgaaggttc aacatcagtt gattgattct gccgccatga
     3541 tcctagttaa gaacccaacc cacctaaacg gtattattat caccagcaat atgtttggtg
     3601 atattatctc cgatgaagcc tccgttatcc caggttcgtt gggtttgttg ccatctgcgt
     3661 cgttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca tgtcacggtt
     3721 ctgcgccaga tttgccaaag aataaggttg atcctatcgc cactatcttg tctgcggcta
     3781 tgatgttgaa attgtcattg aacttgccag aagaaggtaa ggccattgaa gatgcagtta
     3841 aaaaggtttt ggatgcagga atcagaactg gtgatttagg tggttccaat agtaccaccg
     3901 aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt
     3961 ttttatgata tttatacata aactatataa atgaaattca taatagaaac gacacgaaat
     4021 tacaaaatgg aatatgttca taggatatac gaaactatat acgcaatcta catacattta
     4081 tcaagaagga gaaaaaggag gatagtaaag gaatacaggt aagcaaattg atactaatgg
     4141 ctcaacgtga taaggaaaaa gaattgcact ttaacattaa aattgacaag gaataggtca
     4201 ccacacaaaa agttaggtgt aacagaaaat cataaaacta cgattcctaa tttgatattg
     4261 gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca
     4321 tttgatggag tttaagtcaa taccttcttg aa
//
