LOCUS       sequence-79090-e        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast marker swap plasmid for converting KanMX3 to LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Voth WP, Jiang YW, Stillman DJ
  TITLE     New 'marker swap' plasmids for converting selectable markers on 
            budding yeast gene disruptions and plasmids.
  JOURNAL   Yeast. 2003 Aug;20(11):985-93.
  PUBMED    12898713
REFERENCE   2  (bases 1 to 5131)
  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: "Yeast."; 
            date: "2003-08"; volume: "20(11)"; pages: "985-93"
COMMENT     Sequence Label: M4755 KanMX::LEU2 Disruptor Converter
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1101..1298
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1405..1424)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1406..1424)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1511..1528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1682..2270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1762..1781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2441..3232,3233..3301))
                     /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     3064..3083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3302..3406)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3474..3492
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3530..3552)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3652..3671
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3752..3770
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3752..3769
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        3848..4191
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        4215..4620
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             join(4621..5131,1..584)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg
       61 ataaagctaa tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca
      121 agaacgaatt ccctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc
      181 tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata
      241 tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct
      301 tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg
      361 ctccagattt gccaaagaat aaggttgacc ctatcgccac tatcttgtct gctgcaatga
      421 tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa
      481 aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag
      541 tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt
      601 tatgatattt gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac
      661 aaaatggaat atgttcatag ggtagacgaa actatatacg caatctacat acatttatca
      721 agaaggagaa aaaggaggat gtaaaggaat acaggtaagc aaattgatac taatggctca
      781 acgtgataag gaaaaagaat tgcactttaa cattaatatt gacaaggagg agggcaccac
      841 acaaaaagtt aggtgtaaca gaaaatcatg aaactatgat tcctaattta tatattggag
      901 gattttctct aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac ctgaccattt
      961 gatggagttt aagtcaatac cttcttgaac catttcccat aatggtgaaa gttccctcaa
     1021 gaattttact ctgtcagaaa cggccttaac gacgtagtcg actagagtcg acctgcaggc
     1081 atgccctgcc cctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     1141 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     1201 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     1261 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     1321 gccatccagt gtcgaaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     1381 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     1441 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1501 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1561 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1621 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1681 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1741 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1801 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1861 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1921 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1981 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2041 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2101 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     2161 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2221 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2281 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2341 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2401 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2461 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2521 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2581 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2641 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2701 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2761 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2821 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2881 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2941 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3001 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3061 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3121 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3181 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3241 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3301 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3361 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3421 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3481 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3541 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3601 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3661 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     3721 aatacataac cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc
     3781 cagctgaagc ttaaggcgcg ccagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
     3841 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     3901 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     3961 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     4021 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     4081 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     4141 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgggtacc
     4201 cggggatcct ctagaactgt gggaatactc aggtatcgta agatgcaaga gttcgaatct
     4261 cttagcaacc attatttttt tcctcaacat aacgagaaca cacaggggcg ctatcgcaca
     4321 gaatcaaatt cgatgactgg aaattttttg ttaatttcag aggtcgcctg acgcatatac
     4381 ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag agcgccggaa ccggcttttc
     4441 atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa
     4501 tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa
     4561 cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta
     4621 atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca
     4681 gccgaagcca ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat
     4741 ttcgaaaatc atttaattgg tggtgctgct atcgatgcta caggtgtccc acttccagat
     4801 gaggcgctgg aagcctccaa gaaggttgat gccgttttgt taggtgctgt gggtggtcct
     4861 aaatggggta ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt
     4921 caattgtacg ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct
     4981 ccaatcaagc cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga
     5041 ggtatttact ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa
     5101 caatacaccg ttccagaagt gcaaagaatc a
//
