LOCUS       sequence-166088-         997 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            sites X-4, XI-3, and XII-5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 997)
  AUTHORS   Jessop-Fabre MM, Jakociunas T, Stovicek V, Dai Z, Jensen MK, 
            Keasling JD, Borodina I
  TITLE     EasyClone-MarkerFree: A vector toolkit for marker-less integration 
            of genes into Saccharomyces cerevisiae via CRISPR-Cas9.
  JOURNAL   Biotechnol J. 2016 Aug;11(8):1110-7. doi: 10.1002/biot.201600147. 
            Epub 2016 Jun 23.
  PUBMED    27166612
REFERENCE   2  (bases 1 to 997)
  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: 
            "10.1002/biot.201600147"; journalName: "Biotechnol J"; date: 
            "2016-08"; volume: "11"; issue: "8"; pages: "1110-7"
COMMENT     Sequence Label: pCfB3052(gRNA X-4, XI-3, XII-5)
FEATURES             Location/Qualifiers
     source          1..997
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        53..321
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        342..416
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      421..440
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        490..758
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        779..853
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      858..877
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
ORIGIN
        1 ttcgctatta cgccagctgg aatgcgtgcg atagggaaca aaagctggag cttctttgaa
       61 aagataatgt atgattatgc tttcactcat atttatacag aaacttgatg ttttctttcg
      121 agtatataca aggtgattac atgtacgttt gaagtacaac tctagatttt gtagtgccct
      181 cttgggctag cggtaaaggt gcgcattttt tcacacccta caatgttctg ttcaaaagat
      241 tttggtcaaa cgctgtagaa gtgaaagttg gtgcgcatgt ttcggcgttc gaaacttctc
      301 cgcagtgaaa gataaatgat ccgccattca agagcagcaa cgttttagag ctagaaatag
      361 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtggtgc
      421 tttttttgtt ttttatgtct tcgagtcatg taattagtta agtgcaggta gggaacaaaa
      481 gctggagctt ctttgaaaag ataatgtatg attatgcttt cactcatatt tatacagaaa
      541 cttgatgttt tctttcgagt atatacaagg tgattacatg tacgtttgaa gtacaactct
      601 agattttgta gtgccctctt gggctagcgg taaaggtgcg cattttttca caccctacaa
      661 tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg aaagttggtg cgcatgtttc
      721 ggcgttcgaa acttctccgc agtgaaagat aaatgatcat atgtctctaa ttttggaagt
      781 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
      841 caccgagtcg gtggtgcttt ttttgttttt tatgtcttcg agtcatgtaa ttagttaatc
      901 tgtcataggg aacaaaagct ggagcttctt tgaaaagata atgtatgatt atgctttcac
      961 tcatatttat acagaaactt gatgttttct ttcgagt
//
