LOCUS       sequence-158704-         831 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of Cas9 and two gRNA cassettes in S. cerevisiae; Kan 
            Resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 831)
  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 831)
  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-K
FEATURES             Location/Qualifiers
     source          1..831
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        94..362
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        386..459
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      464..483
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      complement(504..751)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     717..735
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(767..787)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
       61 gccagcaacg cgggctaggg ataacagggt aattctttga aaagataatg tatgattatg
      121 ctttcactca tatttataca gaaacttgat gttttctttc gagtatatac aaggtgatta
      181 catgtacgtt tgaagtacaa ctctagattt tgtagtgccc tcttgggcta gcggtaaagg
      241 tgcgcatttt ttcacaccct acaatgttct gttcaaaaga ttttggtcaa acgctgtaga
      301 agtgaaagtt ggtgcgcatg tttcggcgtt cgaaacttct ccgcagtgaa agataaatga
      361 tcggatccac gttctctgcg gccgctttta gagctagaaa tagcaagtta aaataaggct
      421 agtccgttat caacttgaaa aagtggcacc gagtcggtgg tgcttttttt gttttttatg
      481 tctcggcgaa ttgggtaccg gccgcaaatt aaagccttcg agcgtcccaa aaccttctca
      541 agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa
      601 atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata aatagggacc
      661 tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg gggagggcgt
      721 gaatgtaagc gtgacataac taattacatg actcgaggtc gacttaaacc tttctcttct
      781 tctttgggtc agctctagag tcaccaccca attgagacaa gtcaattctg g
//
