LOCUS       sequence-79109-e        4331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast marker swap plasmid for converting KanMX3 to URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4331)
  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 4331)
  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: M4758 KanMX::URA3 Disruptor Converter
FEATURES             Location/Qualifiers
     source          1..4331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        80..423
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        462..677
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             678..1481
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     terminator      1664..1861
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1968..1987)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1969..1987)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2074..2091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2245..2833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2325..2344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3004..3795,3796..3864))
                     /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     3627..3646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3865..3969)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4037..4055
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4093..4115)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4215..4234
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4315..4331,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4315..4331,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttaaggcg cgccagatct gtttagcttg cctcgtcccc
       61 gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt
      121 gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt
      181 ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc
      241 ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca
      301 cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca
      361 tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca
      421 accatgggta cccggggatc cgtcgacctg cagccaagct tttcaattca tcattttttt
      481 tttattcttt tttttgattt cggtttcctt gaaatttttt tgattcggta atctccgaac
      541 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
      601 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacctgca
      661 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
      721 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
      781 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
      841 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
      901 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaagac agaaaatttg
      961 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     1021 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     1081 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     1141 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     1201 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     1261 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagac gcattgggtc
     1321 aacagtatag aaccgtggat gatgtggtct ctacaggatc tgacattatt attgttggaa
     1381 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     1441 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     1501 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     1561 ggaatctcgg tcgtaatgat ttttataatg acgaaaaaaa aaaaattgga aagaaaaagc
     1621 tttcgcgagc tcgacctgca ggcatgccct gcccctaatc agtactgaca ataaaaagat
     1681 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     1741 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     1801 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     1861 gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa acgagctcga attcatcgat
     1921 gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt
     1981 cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     2041 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2101 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     2461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     2701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3961 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4021 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4081 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4141 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4201 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
     4261 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     4321 gtgacactat a
//
