LOCUS       sequence-459803-        3531 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 3531)
  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 3531)
  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: pLS405
FEATURES             Location/Qualifiers
     source          1..3531
                     /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..1627)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1610..1626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1627..2031
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2032..3126
                     /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 gacgcgcgcg
     1561 agctcggtac ccgggatcct ctagagtcga cctgcaggca tgccctagga ctggccgtcg
     1621 ttttacaact gtgggaatac tcaggtatcg taagatgcaa gagttcgagt ctcttagcaa
     1681 ccattatttt tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa
     1741 ttcgatgact ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca
     1801 actgaaaaat tgggagaaaa aggaaaggtg agaggccgga accggctttt catatagaat
     1861 agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta
     1921 aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta
     1981 ccttttacat ttcagcaata tatatatata tttcaaggat ataccattct aatgtctgcc
     2041 cctaagaaga tcgtcgtttt gccaggggac cacgttggtc aagaaatcac agccgaagcc
     2101 attaaggttc ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgagaat
     2161 catttaattg gtggtgctgc tatcgacgct actggtgtcc cacttccaga tgaggctctg
     2221 gaagcctcca agaaggctga tgccgttttg ttaggtgctg tgggtggtcc taaatggggt
     2281 acgggtagtg ttagaccaga acaaggttta ctaaaaatcc gtaaagaact tcaattatac
     2341 gccaatttaa gaccatgtaa ctttgcatcc gattctcttt tagacttatc tccaatcaag
     2401 ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac
     2461 tttggtaaga gaaaggagga cgatggtgat ggtgtcgctt gggatagtga acaatacacc
     2521 gttccagaag tgcaaagaat cacaagaatg gcggctttca tggccctaca acatgagcca
     2581 cctttaccta tttggtcctt agataaagct aatcttttgg cctcatcaag attatggaga
     2641 aaaactgtgg aggaaaccat caagaacgag tttcctacat tgaaggttca acatcagttg
     2701 attgattctg ccgccatgat cctagttaag aacccaaccc acctaaacgg tattattatc
     2761 accagcaata tgtttggtga tattatctcc gatgaagcct ccgttatccc aggttcgttg
     2821 ggtttgttgc catctgcgtc gttggcctct ttgccagaca agaacaccgc atttggtttg
     2881 tacgaaccat gtcacggttc tgcgccagat ttgccaaaga ataaggttga tcctatcgcc
     2941 actatcttgt ctgcggctat gatgttgaaa ttgtcattga acttgccaga agaaggtaag
     3001 gccattgaag atgcagttaa aaaggttttg gatgcaggaa tcagaactgg tgatttaggt
     3061 ggttccaata gtaccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt
     3121 gcttaaaaag attctctttt tttatgatat ttatacataa actatataaa tgaaattcat
     3181 aatagaaacg acacgaaatt acaaaatgga atatgttcat aggatatacg aaactatata
     3241 cgcaatctac atacatttat caagaaggag aaaaaggagg atagtaaagg aatacaggta
     3301 agcaaattga tactaatggc tcaacgtgat aaggaaaaag aattgcactt taacattaaa
     3361 attgacaagg aataggtcac cacacaaaaa gttaggtgta acagaaaatc ataaaactac
     3421 gattcctaat ttgatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa
     3481 acactcaatg acctgaccat ttgatggagt ttaagtcaat accttcttga a
//
