LOCUS       sequence-198965-        5058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of a Cpf1 programming crRNA targetting HIS4 (crHIS4-4.S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5058)
  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 5058)
  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: pUDE714
FEATURES             Location/Qualifiers
     source          1..5058
                     /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            406..1762
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        406..749
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             750..1559
                     /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(817..836)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1565..1762
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1859..1876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2030..2618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2110..2129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2789..3580,3581..3649))
                     /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     3412..3431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3650..3754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4004..4985
                     /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 ttgtagatgc atcttggcta
      301 gcaatgaaca gagaatttct actgttgtag atgtttttta tgtcttatcg tgacgcagtc
      361 ccatgggcca ttacaaactc atgcagacag tcagggaatc ctagcgacat ggaggcccag
      421 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
      481 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
      541 acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc
      601 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
      661 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
      721 tctcacatca catccgaaca taaacaacca tgggtaagga aaagactcac gtttcgaggc
      781 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
      841 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
      901 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
      961 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1021 atgcatggtt actcaccact gcgatccccg gcaaaacagc attccaggta ttagaagaat
     1081 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     1141 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     1201 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     1261 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
     1321 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1381 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     1441 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1501 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     1561 cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta
     1621 tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga
     1681 catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat
     1741 gtgaatgctg gtcgctatac tgtattcacg tagacggata ggtatagcca gacatcagca
     1801 gcatacttcg ggaaccgtag gcgagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1861 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1921 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1981 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2041 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2101 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2161 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2221 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2281 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2341 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2401 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2461 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2521 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2581 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2641 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2701 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2761 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2821 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2881 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2941 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3001 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3061 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3721 gtatttagaa aaataaacaa ataggggttc cgcgcaacta tatgtgaagg catggctatg
     3781 gcacggcaga cattccgcca gatcatcaat aggcaccgcc ctccacgcat ttaagcataa
     3841 acacgcacta tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt
     3901 gcgacgtgaa cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt
     3961 ttcggaaacg ctttgaagtt cctattccga agttcctatt ctgtagaaag tataggaacc
     4021 tcagagcgct tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg
     4081 caccggactg taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa
     4141 aagtatctct ttgctatata tctctgtgct atatccctat ataacctacc catccacctt
     4201 tcgctccttg aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat
     4261 tactctttag acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac
     4321 gaaaatgtaa acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata
     4381 attttctgac caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa
     4441 tccacatcgg tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc
     4501 ttcgctagta atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat
     4561 agacaccaaa gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga
     4621 gactgcatta tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa
     4681 atagcgctct cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta
     4741 aaatagcgct ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg
     4801 aaaaattagc gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt
     4861 tggtaaaata gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt
     4921 gtttgaaaaa ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct
     4981 tcgttggagg gcgtgaacgc atcgctattc agcttgctgc tacagcgcaa cttatctaga
     5041 ttgtatccca tctgaccg
//
