LOCUS       sequence-166081-         996 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 996)
  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 996)
  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..996
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(3..25)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        149..492
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
       61 agcagacaag cccgtcaggg cgcgtcagcg ggctgcattt aaatgatctg tttagcttgc
      121 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
      181 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
      241 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
      301 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
      361 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
      421 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
      481 acataaacaa ccatgggtac cactcttgac gacacggctt accggtaccg caccagtgtc
      541 ccgggggacg ccgaggccat cgaggcactg gatgggtcct tcaccaccga caccgttttc
      601 cgcgtcacca ccaccgggga cggcttcacc ctgcgggagg tgccggtgga cccgcccctg
      661 accaaggtgt tccccgacga cgaatcggac gacgaatcgg acgacgggga ggacggcgac
      721 ccggactccc ggacgttcgt cgcgtacggg gacgacggcg acctggcggg cttcgtggtc
      781 gtctcgtact ccggctggaa ccgccggctg accgtcgagg acatcgaggt cgccccggag
      841 caccgggggc acggggtcgg gcgcgcgttg atggggctcg cgacggagtt cgcccgcgag
      901 cggggcgccg ggcacctctg gctggaggtc accaacgtca acgcaccggc gatccacgcg
      961 taccggcgga tggggttcac cctctgcggc ctggac
//
