LOCUS       sequence-166023-         991 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            site XI-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 991)
  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 991)
  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: pCfB3044(gRNA XI-2)
FEATURES             Location/Qualifiers
     source          1..991
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(79..98)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(103..177)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(198..466)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     terminator      complement(640..837)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggaatg cacgcgatta
       61 actaattaca tgactcgaag acataaaaaa caaaaaaagc accaccgact cggtgccact
      121 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaaccaa
      181 ctgatcaact ggtcaacgat catttatctt tcactgcgga gaagtttcga acgccgaaac
      241 atgcgcacca actttcactt ctacagcgtt tgaccaaaat cttttgaaca gaacattgta
      301 gggtgtgaaa aaatgcgcac ctttaccgct agcccaagag ggcactacaa aatctagagt
      361 tgtacttcaa acgtacatgt aatcaccttg tatatactcg aaagaaaaca tcaagtttct
      421 gtataaatat gagtgaaagc ataatcatac attatctttt caaagaagct ccagcttttg
      481 ttccctatcg cacgcattcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      541 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtaggcggc cgcagctcgt
      601 tttcgacact ggatggcggc gttagtatcg aatcgacagc agtatagcga ccagcattca
      661 catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg ggcagatgat
      721 gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa caactacaat
      781 ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat tgtcagtact
      841 gattaggggc agggcatgct catgtagagc gcctgctcgc cgtccgaggc ggtgccgtcg
      901 tacagggcgg tgtccaggcc gcagagggtg aaccccatcc gccggtacgc gtggatcgcc
      961 ggtgcgttga cgttggtgac ctccagccag a
//
