LOCUS       sequence-272427-        4340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 6 module- LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4340)
  AUTHORS   Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G
  TITLE     COSPLAY: An expandable toolbox for combinatorial and swift 
            generation of expression plasmids in yeast.
  JOURNAL   PLoS One. 2019 Aug 28;14(8):e0220694. doi: 
            10.1371/journal.pone.0220694. eCollection 2019.
  PUBMED    31461461
REFERENCE   2  (bases 1 to 4340)
  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: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pMV16
FEATURES             Location/Qualifiers
     source          1..4340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..68
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     60..77
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     61..77
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        135..542
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             543..1637
                     /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
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(1801..1817)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1801..1817)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1814..1836)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1825..1841
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1849..1855,1856..1873,1874..1879))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1894..1915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2032..2049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2203..2791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2283..2302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2962..3753,3754..3822))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3585..3604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3823..3927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3995..4013
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4051..4073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4173..4192
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
       61 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gatatcggat
      121 cccggtctcg agcaaactgt gggaatactc aggtatcgta agatgcaaga gttcgaatct
      181 cttagcaacc attatttttt tcctcaacat aacgagaaca cacaggggcg ctatcgcaca
      241 gaatcaaatt cgatgactgg aaattttttg ttaatttcag aggtcgcctg acgcatatac
      301 ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag agcgccggaa ccggcttttc
      361 atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa
      421 tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa
      481 cttttcttac cttttacatt tcagcaatat atatatatat atttcaagga tataccattc
      541 taatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt caagaaatca
      601 cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat gtcaagttcg
      661 atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtt ccacttccag
      721 atgaggcgct ggaagcctcc aagaaggctg atgccgtttt gttaggtgct gtgggtggtc
      781 ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc cgtaaagaac
      841 ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt ttagacttat
      901 ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga gaattagtgg
      961 gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct tgggatagtg
     1021 aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc atggccctac
     1081 aacatgagcc accattgcct atttggtcct tggataaagc taatgttttg gcctcttcaa
     1141 gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca ttgaaggttc
     1201 aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc cacctaaatg
     1261 gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc tccgttatcc
     1321 caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac aagaacaccg
     1381 catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag aataaggtca
     1441 accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg aacttgcctg
     1501 aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt atcagaactg
     1561 gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc gaagaagtta
     1621 agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata aactttataa
     1681 atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca tagggtagac
     1741 gaccatagag accgggcccg tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg
     1801 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1861 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2041 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2641 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3001 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc
     3121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3961 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4021 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4081 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4141 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4201 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     4261 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     4321 attacgccag ctggcgaaag
//
