LOCUS       sequence-160425-         996 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of Cas9 and two gRNA cassettes in S. cerevisiae; 
            Nourseothricin Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 996)
  AUTHORS   Generoso WC, Gottardi M, Oreb M, Boles E
  TITLE     Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae.
  JOURNAL   J Microbiol Methods. 2016 Aug;127:203-5. doi: 
            10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17.
  PUBMED    27327211
REFERENCE   2  (bases 1 to 996)
  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: "J Microbiol
            Methods."; date: "2016-08"; volume: "127:203-5. doi"; pages: " 
            10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17"
COMMENT     Sequence Label: pDuRCC-N
FEATURES             Location/Qualifiers
     source          1..996
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(58..77)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        101..369
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        393..466
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      471..490
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(543..561)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    934..981
                     /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)."
ORIGIN
        1 aggagggtat tctgggcctc catgtcgctg attaccctgt tatccctaca ctctcagtac
       61 aatctgctct gatgccgcat agttaagcca gccgggtaat tctttgaaaa gataatgtat
      121 gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag tatatacaag
      181 gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct tgggctagcg
      241 gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt tggtcaaacg
      301 ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg cagtgaaaga
      361 taaatgatcg gatccacgtt ctctgcggcc gcttttagag ctagaaatag caagttaaaa
      421 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtggtgc tttttttgtt
      481 ttttatgtct cggcgaattg ggtgcggccg cgtttgtcac ctttcatagg cctgacgaaa
      541 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagta
      601 tgatccaata tcaaaggaaa tgatagcatt gaaggatgag actaatccaa ttgaggagtg
      661 gcagcatata gaacagctaa agggtagtgc tgaaggaagc atacgatacc ccgcatggaa
      721 tgggataata tcacaggagg tactagacta cctttcatcc tacataaata gacgcatata
      781 agtacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag
      841 gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc
      901 attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc
      961 tctagaaagt ataggaactt cagagcgctt ttgaaa
//
