LOCUS       sequence-166082-         998 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 998)
  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 998)
  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..998
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     209..226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(328..347)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(352..426)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(447..715)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     terminator      complement(765..784)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(789..863)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
       61 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      181 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      241 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggaatgc
      301 acgcgattaa ctaattacat gactcgaaga cataaaaaac aaaaaaagca ccaccgactc
      361 ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta tttctagctc
      421 taaaacatag taggattaga gtttcagatc atttatcttt cactgcggag aagtttcgaa
      481 cgccgaaaca tgcgcaccaa ctttcacttc tacagcgttt gaccaaaatc ttttgaacag
      541 aacattgtag ggtgtgaaaa aatgcgcacc tttaccgcta gcccaagagg gcactacaaa
      601 atctagagtt gtacttcaaa cgtacatgta atcaccttgt atatactcga aagaaaacat
      661 caagtttctg tataaatatg agtgaaagca taatcataca ttatcttttc aaagaagctc
      721 cagcttttgt tccctatgac agattaacta attacatgac tcgaagacat aaaaaacaaa
      781 aaaagcacca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
      841 cttgctattt ctagctctaa aaccaactga tcaactggtc aacgatcatt tatctttcac
      901 tgctgagaag tttcgaacgc cgaaacatgc gcaccaactt tcacttctac agcgtttgac
      961 caaaatcttt tgaacagaac attgtagggt gtgaaaat
//
