LOCUS       sequence-198964-        5102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of a Cpf1 programming crRNA targeting ADE2 and HIS4 
            (crADE2-3 crHIS4-4.S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5102)
  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 5102)
  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: pUDE710
FEATURES             Location/Qualifiers
     source          1..5102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..269
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     gene            450..1806
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        450..793
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             794..1603
                     /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(861..880)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1609..1806
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1903..1920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2074..2662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2154..2173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2833..3624,3625..3693))
                     /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     3456..3475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3694..3798)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4048..5029
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt
       61 ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt
      121 agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt
      181 caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga
      241 aacttctccg cagtgaaaga taaatgatca atttctactg ttgtagatcc ggttgtggta
      301 tatttggtgt ggaaatttct actgttgtag atgcatcttg gctagcaatg aacagagaat
      361 ttctactgtt gtagatgttt tttatgtctt atcgtgacgc agtcccatgg gccattacaa
      421 actcatgcag acagtcaggg aatcctagcg acatggaggc ccagaatacc ctccttgaca
      481 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
      541 atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa
      601 attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
      661 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
      721 tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg
      781 aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat taaattccaa
      841 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
      901 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
      961 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1021 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1081 cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1141 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1201 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1261 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1321 ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     1381 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     1441 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     1501 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     1561 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac tgacaataaa
     1621 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     1681 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     1741 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     1801 atactgtatt cacgtagacg gataggtata gccagacatc agcagcatac ttcgggaacc
     1861 gtaggcgaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3781 acaaataggg gttccgcgca actatatgtg aaggcatggc tatggcacgg cagacattcc
     3841 gccagatcat caataggcac cgccctccac gcatttaagc ataaacacgc actatgccgt
     3901 tcttctcatg tatatatata tacaggcaac acgcagatat aggtgcgacg tgaacagtga
     3961 gctgtatgtg cgcagctcgc gttgcatttt cggaagcgct cgttttcgga aacgctttga
     4021 agttcctatt ccgaagttcc tattctgtag aaagtatagg aacctcagag cgcttttgaa
     4081 aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
     4141 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
     4201 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
     4261 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
     4321 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
     4381 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
     4441 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
     4501 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
     4561 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
     4621 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
     4681 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     4741 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     4801 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     4861 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt
     4921 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     4981 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgttg gagggcgtga
     5041 acgcatcgct attcagcttg ctgctacagc gcaacttatc tagattgtat cccatctgac
     5101 cg
//
