LOCUS       sequence-260825-        9589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S. cerevisiae Episomal plasmid harboring Fncpf1 under the control of
            TDH3p.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9589)
  AUTHORS   Swiat MA, Dashko S, den Ridder M, Wijsman M, van der Oost J, Daran 
            JM, Daran-Lapujade P
  TITLE     FnCpf1: a novel and efficient genome editing tool for Saccharomyces 
            cerevisiae.
  JOURNAL   Nucleic Acids Res. 2017 Dec 1;45(21):12585-12598. doi: 
            10.1093/nar/gkx1007.
  PUBMED    29106617
REFERENCE   2  (bases 1 to 9589)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2017-12-1- 1"; volume: "45"; issue: 
            "21"; pages: "12585-12598"
COMMENT     Sequence Label: pUDE731
FEATURES             Location/Qualifiers
     source          1..9589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    99..146
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      127..1109
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1136..1240
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1241..1309,1310..2101)
                     /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     complement(1459..1478)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2272..2860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2761..2780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3014..3031
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3158..3556
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             7461..7508
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7515..7595
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     terminator      7592..7839
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7608..7626)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(7981..8014)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(9466..9499)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tacaggcaac acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc
       61 gttgcatttt cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc
      121 tattctctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgaaga
      181 cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat
      241 accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc
      301 ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac
      361 attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt
      421 catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag
      481 acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac
      541 tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt
      601 atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca
      661 ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac
      721 ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt
      781 aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa
      841 agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa
      901 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac
      961 aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct
     1021 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
     1081 tctacaaaat gaagcacaga tgcttcgttc aggtggcact tttcggggaa atgtgcgcgg
     1141 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1201 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1261 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1321 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1381 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1441 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     1501 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1561 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1621 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1681 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1741 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     1801 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1861 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1921 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1981 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2041 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2101 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2881 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2941 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3001 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3061 gcctctcgcc tacggttccc gaagtatgct gctgatgtct ggctatacct atccgtctac
     3121 gtgaataccc tcactaaagg gaacaaaagc tggagctcat agcttcaaaa tgtttctact
     3181 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
     3241 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
     3301 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
     3361 caataaaaat ttttatcacg tttctttttc ttgaaaattt tttttttgat ttttttctct
     3421 ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
     3481 tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat
     3541 agcaatctaa tctaagtttt atgagcatct accaggagtt cgtcaacaag tattcactga
     3601 gtaagacact gcggttcgag ctgatcccac agggcaagac actggagaac atcaaggccc
     3661 gaggcctgat tctggacgat gagaagcggg caaaagacta taagaaagcc aagcagatca
     3721 ttgataaata ccaccagttc tttatcgagg aaattctgag ctccgtgtgc atcagtgagg
     3781 atctgctgca gaattactca gacgtgtact tcaagctgaa gaagagcgac gatgacaacc
     3841 tgcagaagga cttcaagtcc gccaaggaca ccatcaagaa acagattagc gagtacatca
     3901 aggactccga aaagtttaaa aatctgttca accagaatct gatcgatgct aagaaaggcc
     3961 aggagtccga cctgatcctg tggctgaaac agtctaagga caatgggatt gaactgttca
     4021 aggctaactc cgatatcact gatattgacg aggcactgga aatcatcaag agcttcaagg
     4081 gatggaccac atactttaaa ggcttccacg agaaccgcaa gaacgtgtac tccagcaacg
     4141 acattcctac ctccatcatc taccgaatcg tcgatgacaa tctgccaaag ttcctggaga
     4201 acaaggccaa atatgaatct ctgaaggaca aagctcccga ggcaattaat tacgaacaga
     4261 tcaagaaaga tctggctgag gaactgacat tcgatatcga ctataagact agcgaggtga
     4321 accagagggt cttttccctg gacgaggtgt ttgaaatcgc caatttcaac aattacctga
     4381 accagtccgg cattactaaa ttcaatacca tcattggcgg gaagtttgtg aacggggaga
     4441 ataccaagcg caagggaatt aacgaataca tcaatctgta tagccagcag atcaacgaca
     4501 aaactctgaa gaaatacaag atgtctgtgc tgttcaaaca gatcctgagt gataccgagt
     4561 ccaagtcttt tgtcattgat aaactggaag atgactcaga cgtggtcact accatgcaga
     4621 gcttttatga gcagatcgcc gctttcaaga cagtggagga aaaatctatt aaggaaactc
     4681 tgagtctgct gttcgatgac ctgaaagccc agaagctgga cctgagtaag atctacttca
     4741 aaaacgataa gagtctgaca gacctgtcac agcaggtgtt tgatgactat tccgtgattg
     4801 ggaccgccgt cctggagtac attacacagc agatcgctcc aaagaacctg gataatccct
     4861 ctaagaaaga gcaggaactg atcgctaaga aaaccgagaa ggcaaaatat ctgagtctgg
     4921 aaacaattaa gctggcactg gaggagttca acaagcacag ggatattgac aaacagtgcc
     4981 gctttgagga aatcctggcc aacttcgcag ccatccccat gatttttgat gagatcgccc
     5041 agaacaaaga caatctggct cagatcagta ttaagtacca gaaccagggc aagaaagacc
     5101 tgctgcaggc ttcagcagaa gatgacgtga aagccatcaa ggatctgctg gaccagacca
     5161 acaatctgct gcacaagctg aaaatcttcc atattagtca gtcagaggat aaggctaata
     5221 tcctggataa agacgaacac ttctacctgg tgttcgagga atgttacttc gagctggcaa
     5281 acattgtccc cctgtataac aagattagga actacatcac acagaagcct tactctgacg
     5341 agaagtttaa actgaacttc gaaaatagta ccctggccaa cgggtgggat aagaacaagg
     5401 agcctgacaa cacagctatc ctgttcatca aggatgacaa gtactatctg ggagtgatga
     5461 ataagaaaaa caataagatc ttcgatgaca aagccattaa ggagaacaaa ggggaaggat
     5521 acaagaaaat cgtgtataag ctgctgcccg gcgcaaataa gatgctgcct aaggtgttct
     5581 tcagcgccaa gagtatcaaa ttctacaacc catccgagga catcctgcgg attagaaatc
     5641 actcaacaca tactaagaac gggagccccc agaagggata tgagaaattt gagttcaaca
     5701 tcgaggattg caggaagttt attgacttct acaagcagag catctccaaa caccctgaat
     5761 ggaaggattt tggcttccgg ttttccgaca cacagagata taactctatc gacgagttct
     5821 accgcgaggt ggaaaatcag gggtataagc tgacttttga gaacatttct gaaagttaca
     5881 tcgacagcgt ggtcaatcag ggaaagctgt acctgttcca gatctataac aaagattttt
     5941 cagcatacag caagggcaga ccaaacctgc atacactgta ctggaaggcc ctgttcgatg
     6001 agaggaatct gcaggacgtg gtctataaac tgaacggaga ggccgaactg ttttaccgga
     6061 agcagtctat tcctaagaaa atcactcacc cagctaagga ggccatcgct aacaagaaca
     6121 aggacaatcc taagaaagag agcgtgttcg aatacgatct gattaaggac aagcggttca
     6181 ccgaagataa gttctttttc cattgtccaa tcaccattaa cttcaagtca agcggcgcta
     6241 acaagttcaa cgacgagatc aatctgctgc tgaaggaaaa agcaaacgat gtgcacatcc
     6301 tgagcattga ccgaggagag cggcatctgg cctactatac cctggtggat ggcaaaggga
     6361 atatcattaa gcaggataca ttcaacatca ttggcaatga ccggatgaaa accaactacc
     6421 acgataaact ggctgcaatc gagaaggata gagactcagc taggaaggac tggaagaaaa
     6481 tcaacaacat taaggagatg aaggaaggct atctgagcca ggtggtccat gagattgcaa
     6541 agctggtcat cgaatacaat gccattgtgg tgttcgagga tctgaacttc ggctttaaga
     6601 gggggcgctt taaggtggaa aaacaggtct atcagaagct ggagaaaatg ctgatcgaaa
     6661 agctgaatta cctggtgttt aaagataacg agttcgacaa gaccggaggc gtcctgagag
     6721 cctaccagct gacagctccc tttgaaactt tcaagaaaat gggaaaacag acaggcatca
     6781 tctactatgt gccagccgga ttcacttcca agatctgccc cgtgaccggc tttgtcaacc
     6841 agctgtaccc taaatatgag tcagtgagca agtcccagga atttttcagc aagttcgata
     6901 agatctgtta taatctggac aaggggtact tcgagttttc cttcgattac aagaacttcg
     6961 gcgacaaggc cgctaagggg aaatggacca ttgcctcctt cggatctcgc ctgatcaact
     7021 ttcgaaattc cgataaaaac cacaattggg acactaggga ggtgtaccca accaaggagc
     7081 tggaaaagct gctgaaagac tactctatcg agtatggaca tggcgaatgc atcaaggcag
     7141 ccatctgtgg cgagagtgat aagaaatttt tcgccaagct gacctcagtg ctgaatacaa
     7201 tcctgcagat gcggaactca aagaccggga cagaactgga ctatctgatt agccccgtgg
     7261 ctgatgtcaa cggaaacttc ttcgacagca gacaggcacc caaaaatatg cctcaggatg
     7321 cagacgccaa cggggcctac cacatcgggc tgaagggact gatgctgctg ggccggatca
     7381 agaacaatca ggaggggaag aagctgaacc tggtcattaa gaacgaggaa tacttcgagt
     7441 ttgtccagaa tagaaataac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
     7501 agaaaaaggg atcctaccca tacgatgttc cagattacgc ttatccctac gacgtgcctg
     7561 attatgcata cccatatgat gtccccgact atgcctaaat tagttatgtc acgcttacat
     7621 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     7681 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     7741 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     7801 cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gcccaccttt cgagaggacg
     7861 atgcccgtgt ctaaatgatt cgaccagcct aagaatgttc aacggctggt cgaatcattt
     7921 agacacgggc atcgtcctct cgaaaggtgg ctaggccact agtggatctg atatcaccta
     7981 ataacttcgt atagcataca ttatacgaag ttatattaag ggttctcgag agctcgtttt
     8041 atttaggttc tatcgaggag aaaaagcgac aagaagagat agaccatgga taaatgatta
     8101 tgttctaaac actcctcaga agctcatcga actgtcatcc tgcgtgaaga ttaaaatcca
     8161 acttagaaat ttcgagctta cggagacaat catatgggag aagcaattgg aagatagaaa
     8221 aaaggtactc ggtacataaa tatatgtgat tctgggtaga agatcggtct gcattggatg
     8281 gtggtaacgc atttttttac acacattact tgcctcgagc atcaaatggt ggttattcgt
     8341 ggatctatat cacgtgattt gcttaagaat tgtcgttcat ggtgacactt ttagctttga
     8401 catgattaag ctcatctcaa ttgatgttat ctaaagtcat ttcaactatc taagatgtgg
     8461 ttgtgattgg gccattttgt gaaagccagt acgccagcgt caatacactc ccgtcaatta
     8521 gttgcaccat gtccacaaaa tcatatacca gtagagctga gactcatgca agtccggttg
     8581 catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct tctcttgacg
     8641 ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac atttgccttt
     8701 tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc gttccattga
     8761 aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc gccgatatcg
     8821 gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa tggtctgata
     8881 tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa caaggtgcgc
     8941 aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct tccaagggtt
     9001 ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt gataaagatt
     9061 tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg tttgattggc
     9121 taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt cagcagtaca
     9181 gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc agaggacttt
     9241 tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct ggatgggaag
     9301 cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat agaacaaatc
     9361 acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc aacttcatct
     9421 tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac ttcgtatagc
     9481 atacattata cgaagttata ttaagggttg tcgacctgca gcgtacgaag gtgcctattg
     9541 atgatctggc ggaatgtctg ccgtgccata gccatgcctt cacatatag
//
