LOCUS       sequence-79100-e        6042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast marker swap plasmid for converting Trp1 to KanMX3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  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 6042)
  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: M3925 trp1::KanMX3 Disruptor Converter
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..138
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     gene            complement(1052..2408)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(1052..1249)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1255..2064)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1978..1997
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2065..2408)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(3426..3442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3426..3442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3439..3461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3450..3466
                     /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(3474..3480,3481..3498,3499..3504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3519..3540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3657..3674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3828..4416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3908..3927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4587..5378,5379..5447))
                     /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     5210..5229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5448..5552)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5620..5638
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5676..5698)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5798..5817
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6011..6033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6025..6042
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6026..6042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcccgg ggatccggtg attgattgag caagctaatt cggtcgaaaa aagaaaagga
       61 gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac
      121 acaaaggcag cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg
      181 aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac
      241 ttgctgggta ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg
      301 aaaatttcaa gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt
      361 ggcgtgtttc gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt
      421 gaatcatcga tgaattcgag ctcgtttacc gtttaaacgt acaaagatgg catcgaccta
      481 aacaccatag aatggtcggg ccattgactg cctatttaac gccaacgttg cgagatatac
      541 actcgaaatg tgtgatgtag atgtaaacag acggctatat gaatgccact agccggttat
      601 tttacttcta gttaatacat atttcctaga attttttacg tgcataagag tagaatagtc
      661 taaagagaag ggtcctcgag gctagctcag accagctgcg gattgcctgc gcaaggcttc
      721 cttaactgca agcgcaatgg cgtcgccaac ctccgaagtg gagctcgagc cgagcaggtc
      781 gcccgttctg actcccgagt ccaacacctc ctgcactgcc tgctcgaccg cctcgccggc
      841 agcaaccatg ttcaacgaca acttcagcat catggcagca gagaggatgc atccgatcgg
      901 gttcgccttc ccggcgggca gatcgggcgc agagccgtgg cagggctcgt agaggccaaa
      961 ggcgctcttg ctatccggca aagacgcgag cgaggccgat ggcagcaacc ctagtcgaca
     1021 ctggatggcg gcgttagtat cgaatcgaca gcagtatagc gaccagcatt cacatacgat
     1081 tgacgcatga tattactttc tgcgcactta acttcgcatc tgggcagatg atgtcgaggc
     1141 gaaaaaaaat ataaatcacg ctaacatttg attaaaatag aacaactaca atataaaaaa
     1201 actatacaaa tgacaagttc ttgaaaacaa gaatcttttt attgtcagta ctgattagaa
     1261 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     1321 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     1381 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     1441 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     1501 cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt
     1561 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     1621 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     1681 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     1741 taatacctgg aatgctgttt tgccggggat cgcagtggtg agtaaccatg catcatcagg
     1801 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     1861 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     1921 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     1981 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     2041 aacgtgagtc ttttccttac ccatggttgt ttatgttcgg atgtgatgtg agaactgtat
     2101 cctagcaaga ttttaaaagg aagtatatga aagaagaacc tcagtggcaa atcctaacct
     2161 tttatatttc tctacagggg cgcggcgtgg ggacaattca acgcgtctgt gaggggagcg
     2221 tttccctgct cgcaggtctg cagcgaggag ccgtaatttt tgcttcgcgc cgtgcggcca
     2281 tcaaaatgta tggatgcaaa tgattataca tggggatgta tgggctaaat gtacgggcga
     2341 cagtcacatc atgcccctga gctgcgcacg tcaagactgt caaggagggt attctgggcc
     2401 tccatgtcgc tggccgggtg acccggcggg gacgaggcaa gctgtacaaa gatggcatcg
     2461 acctaaacac catagaatgg tcgggccatt gactgcctat ttaacgccaa cgttgcgaga
     2521 tatacactcg aaatgtgtga tgtagatgta aacagacggc tatatgaatg ccactagccg
     2581 gttattttac tcctagttaa tacatatttc ctagaatttt ttacgtgcat aagagtagaa
     2641 tagtctaaag agaagggtcc tcgaggctag ctcagaccag ctgcggattg cctgcgcaag
     2701 gcttccttaa ctgcaagcgc aatggcgtcg ccaacctccg aagtggagct cgagccgagc
     2761 aggtcgcccg ttctgactcc cgagtccaac acctcctgca ctgcctgctc gaccgcctcg
     2821 ccggcagcaa ccatgttcaa cgacaacttc agcatcatgg cagcagagag gatgcatccg
     2881 atcgggttcg ccttcccggc gggcagatct ggcgcgcctt aattaaccca tcgtccaact
     2941 gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag
     3001 actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc
     3061 gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc
     3121 gatttctgac tgggttggaa ggcaagagag ccccgaaagc ttacatttta tgttagctgg
     3181 tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga
     3241 tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt
     3301 caaaaatgct aagaaatagg ttattactga gtagtattta tttaagtatt gtttgtgcac
     3361 ttgcccttgc tcaatcaatc accggatccg tcgacctgca gccaagctag cttggcgtaa
     3421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3541 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3601 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3661 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3721 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4021 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4261 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4381 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4441 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4501 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4561 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4621 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4681 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4741 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4801 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4861 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4921 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4981 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5041 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5101 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5161 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5221 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5281 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5341 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5401 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5461 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5521 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5581 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5641 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5701 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5761 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     5821 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     5881 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5941 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6001 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gt
//
