LOCUS       sequence-79106-e        6295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast marker swap plasmid for converting URA3 to KanMX3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  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 6295)
  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: M3927 ura3::KanMX3 Disruptor Converter
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1231..2587
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1231..1574
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1575..2384
                     /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     complement(1642..1661)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2390..2587
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(3396..3611)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3679..3695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3679..3695)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3692..3714)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3703..3719
                     /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(3727..3733,3734..3751,3752..3757))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3772..3793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3910..3927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4081..4669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4161..4180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4840..5631,5632..5700))
                     /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     5463..5482
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5701..5805)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5873..5891
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5929..5951)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6051..6070
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6264..6286
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6278..6295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6279..6295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcccgg ggatccggtg attgattgag caagctgggt aataactgat ataattaaat
       61 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
      121 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
      181 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
      241 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
      301 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
      361 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
      421 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
      481 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
      541 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
      601 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
      661 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
      721 agtcaagatg atctgcccgc cgggaaggcg aacccgatcg gatgcatcct ctctgctgcc
      781 atgatgctga agttgtcgtt gaacatggtt gctgccggcg aggcggtcga gcaggcagtg
      841 caggaggtgt tggactcggg agtcagaacg ggcgacctgc tcggctcgag ctccacttcg
      901 gaggttggcg acgccattgc gcttgcagtt aaggaagcct tgcgcaggca atccgcagct
      961 ggtctgagct agcctcgagg acccttctct ttagactatt ctactcttat gcacgtaaaa
     1021 aattctagga aatatgtatt aactaggagt aaaataaccg gctagtggca ttcatatagc
     1081 cgtctgttta catctacatc acacatttcg agtgtatatc tcgcaacgtt ggcgttaaat
     1141 aggcagtcaa tggcccgacc attctatggt gtttaggtcg atgccatctt tgtacagctt
     1201 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1261 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1321 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1381 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1441 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1501 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     1561 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
     1621 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1681 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1741 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1801 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1861 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1921 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1981 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2041 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2101 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2161 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2221 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2281 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2341 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     2401 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     2461 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     2521 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     2581 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgactaggg ttgctgccat
     2641 cggcctcgct cgcgtctttg ccggatagca agagcgcctt tggcctctac gagccctgcc
     2701 acggctctgc gcccgatctg cccgccggga aggcgaaccc gatcggatgc atcctctctg
     2761 ctgccatgat gctgaagttg tcgttgaaca tggttgctgc cggcgaggcg gtcgagcagg
     2821 cagtgcagga ggtgttggac tcgggagtca gaacgggcga cctgctcggc tcgagctcca
     2881 cttcggaggt tggcgacgcc attgcgcttg cagttaagga agccttgcgc aggcaatccg
     2941 cagctggtct gagctagcct cgaggaccct tctctttaga ctattctact cttatgcacg
     3001 taaaaaattc taggaaatat gtattaacta gaagtaaaat aaccggctag tggcattcat
     3061 atagccgtct gtttacatct acatcacaca tttcgagtgt atatctcgca acgttggcgt
     3121 taaataggca gtcaatggcc cgaccattct atggtgttta ggtcgatgcc atctttgtac
     3181 gtttaaacgg taaacgagct cgaattatcc acatgtgttt ttagtaaaca aattttggga
     3241 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     3301 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     3361 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     3421 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     3481 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     3541 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     3601 gatgaattga aaagctagct tgctcaatca atcaccggat ccgtcgacct gcagccaagc
     3661 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3721 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3781 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3841 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3901 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3961 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4021 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4081 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4141 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4201 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4261 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4321 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4381 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4441 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4501 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4561 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4621 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4681 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4741 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4801 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4861 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4921 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4981 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5041 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5101 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5161 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5221 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5281 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5341 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5401 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5461 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5521 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5581 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5641 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5701 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5761 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5821 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5881 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5941 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6001 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6061 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6121 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6181 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6241 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagt
//
