LOCUS       sequence-198968-        5190 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expression of a Cpf1 programming crRNA targetting CAN1, HIS4, PDR12 
            and ADE2 (crCAN1-4.crHIS4-4.crPDR12-3.crADE2-3.S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5190)
  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 5190)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pUDE735
FEATURES             Location/Qualifiers
     source          1..5190
                     /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            538..1894
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        538..881
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             882..1691
                     /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(949..968)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1697..1894
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1991..2008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2162..2750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2242..2261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2921..3712,3713..3781))
                     /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     3544..3563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3782..3886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4136..5117
                     /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 acatatcttc
      301 aacgctgtta tctaatttct actgttgtag atgcatcttg gctagcaatg aacagagaat
      361 ttctactgtt gtagatgcac aaagaatcaa tatgggtgtc aaatttctac tgttgtagat
      421 ccggttgtgg tatatttggt gtggaaattt ctactgttgt agatgttttt tatgtcttat
      481 cgtgacgcag tcccatgggc cattacaaac tcatgcagac agtcagggaa tcctagcgac
      541 atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt
      601 gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat
      661 tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag
      721 ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat
      781 ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg
      841 ctaggataca gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc
      901 acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
      961 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     1021 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     1081 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     1141 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg
     1201 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     1261 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     1321 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     1381 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     1441 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     1501 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     1561 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     1621 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     1681 agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     1741 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     1801 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     1861 cgtcaatcgt atgtgaatgc tggtcgctat actgtattca cgtagacgga taggtatagc
     1921 cagacatcag cagcatactt cgggaaccgt aggcgagagg cggtttgcgt attgggcgct
     1981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2341 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2581 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2701 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2761 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2821 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2881 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2941 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3001 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3061 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3121 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3181 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3241 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3301 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3361 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3421 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3481 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3541 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3601 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3661 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3721 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3781 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3841 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaac tatatgtgaa
     3901 ggcatggcta tggcacggca gacattccgc cagatcatca ataggcaccg ccctccacgc
     3961 atttaagcat aaacacgcac tatgccgttc ttctcatgta tatatatata caggcaacac
     4021 gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt tgcattttcg
     4081 gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc gaagttccta ttctgtagaa
     4141 agtataggaa cctcagagcg cttttgaaaa ccaaaagcgc tctgaagacg cactttcaaa
     4201 aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac cgcttccaca
     4261 aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct atataaccta
     4321 cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat tttttatgtt
     4381 tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca tagagtgaat
     4441 cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac aaaatagaag
     4501 aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt tctgttcaca
     4561 aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat cttgaaaaaa
     4621 tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg ctttttttat
     4681 ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct acagtgcaaa
     4741 aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc
     4801 tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat
     4861 tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     4921 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca
     4981 cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag
     5041 ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga
     5101 agcacagatg cttcgttgga gggcgtgaac gcatcgctat tcagcttgct gctacagcgc
     5161 aacttatcta gattgtatcc catctgaccg
//
