LOCUS       sequence-166083-         901 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            sites X-3, XI-2, and XII-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 901)
  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 901)
  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: pCfB3051(gRNA X-3 XI-2 XII-2)
FEATURES             Location/Qualifiers
     source          1..901
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        56..324
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        345..419
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      424..443
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        493..761
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        782..856
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      861..880
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
ORIGIN
        1 ctcttcgcta ttacgccagc tggaatgcgt gcgataggga acaaaagctg gagcttcttt
       61 gaaaagataa tgtatgatta tgctttcact catatttata cagaaacttg atgttttctt
      121 tcgagtatat acaaggtgat tacatgtacg tttgaagtac aactctagat tttgtagtgc
      181 cctcttgggc tagcggtaaa ggtgcgcatt ttttcacacc ctacaatgtt ctgttcaaaa
      241 gattttggtc aaacgctgta gaagtgaaag ttggtgcgca tgtttcggcg ttcgaaactt
      301 ctccgcagtg aaagataaat gatcctaatg tgtccgcgtt tctagtttta gagctagaaa
      361 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgg
      421 tgcttttttt gttttttatg tcttcgagtc atgtaattag ttaagtgcag gtagggaaca
      481 aaagctggag cttctttgaa aagataatgt atgattatgc tttcactcat atttatacag
      541 aaacttgatg ttttctttcg agtatataca aggtgattac atgtacgttt gaagtacaac
      601 tctagatttt gtagtgccct cttgggctag cggtaaaggt gcgcattttt tcacacccta
      661 caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa gtgaaagttg gtgcgcatgt
      721 ttcggcgttc gaaacttctc agcagtgaaa gataaatgat cgttgaccag ttgatcagtt
      781 ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
      841 tggcaccgag tcggtggtgc tttttttgtt ttttatgtct tcgagtcatg taattagtta
      901 a
//
