LOCUS       sequence-408526-        9495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Episomal yeast plasmid expressing the Ercas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9495)
  AUTHORS   Bennis NX, Anderson JP, Kok SMC, Daran JG
  TITLE     Expanding the genome editing toolbox of Saccharomyces cerevisiae 
            with the endonuclease ErCas12a.
  JOURNAL   FEMS Yeast Res. 2023 Jan 4;23:foad043. doi: 10.1093/femsyr/foad043.
  PUBMED    37791490
REFERENCE   2  (bases 1 to 9495)
  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: "FEMS Yeast 
            Res."; date: "2023-01-4"; volume: "23:foad043. doi"; pages: " 
            10.1093/femsyr/foad043"
COMMENT     Sequence Label: pUDE1093
FEATURES             Location/Qualifiers
     source          1..9495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4512..4532
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     gene            5012..6367
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             5356..6165
                     /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(5423..5442)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6170..6367
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      6372..7354
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(7385..8176)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      8606..9194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9095..9114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggtgagt aaggaaagag tgaggaacta tcgcatacct gcatttaaag atgccgattt
       61 gggcgcgaat cctttatttt ggcttcaccc tcatactatt atcagggcca gaaaaaggaa
      121 gtgtttccct ccttcttgaa ttgatgttac cctcataaag cacgtggcct cttatcgaga
      181 aagaaattac cgtcgctcgt gatttgtttg caaaaagaac aaaactgaaa aaacccagac
      241 acgctcgact tcctgtcatc ctattgattg cagcttccaa tttcgtcaca caacaaggtc
      301 ctagcgacgg ctcacaggtt ttgtaacaag caatcgaagg ttctggaatg gcgggaaagg
      361 gtttagtacc acatgctatg atgcccactg tgatctccag agcaaagttc gttcgatcgt
      421 actgttactc tctctctttc aaacagaatt gtccgaatcg tgtgacaaca acagcctgtt
      481 ctcacacact cttttcttct aaccaagggg gtggtttagt ttagtagaac ctcgtgaaac
      541 ttacatttac atatatataa acttgcataa attggtcaat gcaagaaata catatttggt
      601 cttttctaat tcgtagtttt tcaagttctt agatgctttc tttttctctt ttttacagat
      661 catcaaggaa gtaattatct actttttaca acaaatataa aacaagatct atgaacaacg
      721 gtactaacaa cttccaaaac ttcatcggta tctcttcttt gcaaaagact ttgagaaacg
      781 ctttgatccc aactgaaact actcaacaat tcatcgttaa gaacggtatc atcaaggaag
      841 acgaattgag aggtgaaaac agacaaatct tgaaggacat catggacgac tactacagag
      901 gtttcatctc tgaaactttg tcttctatcg acgacatcga ctggacttct ttgttcgaaa
      961 agatggaaat ccaattgaag aacggtgaca acaaggacac tttgatcaag gaacaaactg
     1021 aatacagaaa ggctatccac aagaagttcg ctaacgacga cagattcaag aacatgttct
     1081 ctgctaagtt gatctctgac atcttgccag aattcgttat ccacaacaac aactactctg
     1141 cttctgaaaa ggaagaaaag actcaagtta tcaagttgtt ctctagattc gctacttctt
     1201 tcaaggacta cttcaagaac agagctaact gtttctctgc tgacgacatc tcttcttctt
     1261 cttgtcacag aatcgttaac gacaacgctg aaatcttctt ctctaacgct ttggtttaca
     1321 gaagaatcgt taagtctttg tctaacgacg acatcaacaa gatctctggt gacatgaagg
     1381 actctttgaa ggaaatgtct ttggaagaaa tctactctta cgaaaagtac ggtgaattca
     1441 tcactcaaga aggtatctct ttctacaacg acatctgtgg taaggttaac tctttcatga
     1501 acttgtactg tcaaaagaac aaggaaaaca agaacttgta caagttgcaa aagttgcaca
     1561 agcaaatctt gtgtatcgct gacacttctt acgaagttcc atacaagttc gaatctgacg
     1621 aagaagttta ccaatctgtt aacggtttct tggacaacat ctcttctaag cacatcgttg
     1681 aaagattgag aaagatcggt gacaactaca acggttacaa cttggacaag atctacatcg
     1741 tttctaagtt ctacgaatct gtttctcaaa agacttacag agactgggaa actatcaaca
     1801 ctgctttgga aatccactac aacaacatct tgccaggtaa cggtaagtct aaggctgaca
     1861 aggttaagaa ggctgttaag aacgacttgc aaaagtctat cactgaaatc aacgaattgg
     1921 tttctaacta caagttgtgt tctgacgaca acatcaaggc tgaaacttac atccacgaaa
     1981 tctctcacat cttgaacaac ttcgaagctc aagaattgaa gtacaaccca gaaatccact
     2041 tggttgaatc tgaattgaag gcttctgaat tgaagaacgt tttggacgtt atcatgaacg
     2101 ctttccactg gtgttctgtt ttcatgactg aagaattggt tgacaaggac aacaacttct
     2161 acgctgaatt ggaagaaatc tacgacgaaa tctacccagt tatctctttg tacaacttgg
     2221 ttagaaacta cgttactcaa aagccatact ctactaagaa gatcaagttg aacttcggta
     2281 tcccaacttt ggctgacggt tggtctaagt ctaaggaata ctctaacaac gctatcatct
     2341 tgatgagaga caacttgtac tacttgggta tcttcaacgc taagaacaag ccagacaaga
     2401 agatcatcga aggtaacact tctgaaaaca agggtgacta caagaagatg atctacaact
     2461 tgttgccagg tccaaacaag atgatcccaa aggttttctt gtcttctaag actggtgttg
     2521 aaacttacaa gccatctgct tacatcttgg aaggttacaa gcaaaacaag cacatcaagt
     2581 cttctaagga cttcgacatc actttctgtc acgacttgat cgactacttc aagaactgta
     2641 tcgctatcca cccagaatgg aagaacttcg gtttcgactt ctctgacact tctacttacg
     2701 aagacatctc tggtttctac agagaagttg aattgcaagg ttacaagatc gactggactt
     2761 acatctctga aaaggacatc gacttgttgc aagaaaaggg tcaattgtac ttgttccaaa
     2821 tctacaacaa ggacttctct aagaagtcta ctggtaacga caacttgcac actatgtact
     2881 tgaagaactt gttctctgaa gaaaacttga aggacatcgt tttgaagttg aacggtgaag
     2941 ctgaaatctt cttcagaaag tcttctatca agaacccaat catccacaag aagggttcta
     3001 tcttggttaa cagaacttac gaagctgaag aaaaggacca attcggtaac atccaaatcg
     3061 ttagaaagaa catcccagaa aacatctacc aagaattgta caagtacttc aacgacaagt
     3121 ctgacaagga attgtctgac gaagctgcta agttgaagaa cgttgttggt caccacgaag
     3181 ctgctactaa catcgttaag gactacagat acacttacga caagtacttc ttgcacatgc
     3241 caatcactat caacttcaag gctaacaaga ctggtttcat caacgacaga atcttgcaat
     3301 acatcgctaa ggaaaaggac ttgcacgtta tcggtatcga cagaggtgaa agaaacttga
     3361 tctacgtttc tgttatcgac acttgtggta acatcgttga acaaaagtct ttcaacatcg
     3421 ttaacggtta cgactaccaa atcaagttga agcaacaaga aggtgctaga caaatcgcta
     3481 gaaaggaatg gaaggaaatc ggtaagatca aggaaatcaa ggaaggttac ttgtctttgg
     3541 ttatccacga aatctctaag atggttatca agtacaacgc tatcatcgct atggaagact
     3601 tgtcttacgg tttcaagaag ggtagattca aggttgaaag acaagtttac caaaagttcg
     3661 aaactatgtt gatcaacaag ttgaactact tggttttcaa ggacatctct atcactgaaa
     3721 acggtggttt gttgaagggt taccaattga cttacatccc agacaagttg aagaacgttg
     3781 gtcaccaatg tggttgtatc ttctacgttc cagctgctta cacttctaag atcgacccaa
     3841 ctactggttt cgttaacatc ttcaagttca aggacttgac tgttgacgct aagagagaat
     3901 tcatcaagaa gttcgactct atcagatacg actctgaaaa gaacttgttc tgtttcactt
     3961 tcgactacaa caacttcatc actcaaaaca ctgttatgtc taagtcttct tggtctgttt
     4021 acacttacgg tgttagaatc aagagaagat tcgttaacgg tagattctct aacgaatctg
     4081 acactatcga catcactaag gacatggaaa agactttgga aatgactgac atcaactgga
     4141 gagatggtca cgacttgaga caagacatca tcgactacga aatcgttcaa cacatcttcg
     4201 aaatcttcag attgactgtt caaatgagaa actctttgtc tgaattggaa gacagagact
     4261 acgacagatt gatctctcca gttttgaacg aaaacaacat cttctacgac tctgctaagg
     4321 ctggtgacgc tttgccaaag gacgctgacg ctaacggtgc ttactgtatc gctttgaagg
     4381 gtttgtacga aatcaagcaa atcactgaaa actggaagga agacggtaag ttctctagag
     4441 acaagttgaa gatctctaac aaggactggt tcgacttcat ccaaaacaag agatacttga
     4501 gcagggctga ccccaagaag aagaggaagg tgtaaatcct tttgtataac taaataatat
     4561 tggaaactaa atacgaatac ccaaattttt tatctaaatt ttgccgaaag attaaaatct
     4621 gcagagatat ccgaaacagg taaatggatg tttcaatccc tgtagtcagt caggaaccca
     4681 tattatatta cagtattagt cgccgcttag gcacgccttt aattagcaaa atcaaacctt
     4741 aagtgcatat gccgtataag ggaaactcaa agaactggca tcgcaaaaat gaaaagctgc
     4801 caatgagacg acggggtcat cacggctcat catgcgccaa acaaatgtgt gcaatacacg
     4861 ctcggatgac tgcatgatga ccgcactgac tggggacagc agatccacct aagcctgtga
     4921 gagaagcaga cacccgacag atcaaggcag ttaactagtg cactgcagta cagtttagct
     4981 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
     5041 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
     5101 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
     5161 aaattacggc tcctcgctcc agacctgcga gcagggaaac gctcccctca cagacgcgtt
     5221 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
     5281 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
     5341 cgaacataaa caaaaatggg taaggaaaag actcacgttt cgaggccgcg attaaattcc
     5401 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     5461 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     5521 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     5581 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     5641 accactgcga tccccggcaa aacagcattc caggtattag aagaatatcc tgattcaggt
     5701 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     5761 aattgtcctt ttaacagcga tcgcgtattt cgtctggctc aggcgcaatc acgaatgaat
     5821 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     5881 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
     5941 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     6001 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     6061 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     6121 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaaagtaa ctgacaataa
     6181 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     6241 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     6301 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     6361 tatactggag taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     6421 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     6481 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     6541 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     6601 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     6661 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     6721 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     6781 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     6841 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     6901 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     6961 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     7021 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     7081 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     7141 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     7201 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     7261 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt
     7321 ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg
     7381 ccgcttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca
     7441 actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt
     7501 caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat
     7561 ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta
     7621 catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta
     7681 gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta
     7741 ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg
     7801 tggctggctc gaagataccc gcaagaatgt cattgcgctg ccattctcca aattgcagtt
     7861 cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta
     7921 cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca
     7981 aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac
     8041 tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg
     8101 gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga
     8161 tcaccgcttc cctcataatg tttaactttg ttttagggcg actgccctgc tgcgtaacat
     8221 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     8281 cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa accgccactg
     8341 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     8401 acttgcatta cagcttacga accgaacagg cttatgtcca ctgggttcgt gccttcatcc
     8461 gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag gcatttctgt
     8521 cctggctggt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     8581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     8761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     9001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     9061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     9121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     9181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     9241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
     9301 cccctgaatt cgcatctaga tggtagagcc acaaacagcc ggtacaagca acgatctcca
     9361 ggaccatctg aatcatgcgc ggatgacacg aactcacgac ggcgatcaca gacattaacc
     9421 cacagtacag acactgcgac aacgtggcaa ttcgtcgcaa taccgtctca ctgaactggc
     9481 cgataattgc agacg
//
