LOCUS       sequence-166069-        1001 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            site X-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1001)
  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 1001)
  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: pCfB3042(gRNA X-4)
FEATURES             Location/Qualifiers
     source          1..1001
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(10..32)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        156..499
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
       61 tgccgggagc agacaagccc gtcagggcgc gtcagcgggc tgcatttaaa tgatctgttt
      121 agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc
      181 ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc
      241 ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa
      301 gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg
      361 cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac
      421 tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca
      481 catccgaaca taaacaacca tgggtaccac tcttgacgac acggcttacc ggtaccgcac
      541 cagtgtcccg ggggacgccg aggccatcga ggcactggat gggtccttca ccaccgacac
      601 cgttttccgc gtcaccacca ccggggacgg cttcaccctg cgggaggtgc cggtggaccc
      661 gcccctgacc aaggtgttcc ccgacgacga atcggacgac gaatcggacg acggggagga
      721 cggcgacccg gactcccgga cgttcgtcgc gtacggggac gacggcgacc tggcgggctt
      781 cgtggtcgtc tcgtactccg gctggaaccg ccggctgacc gtcgaggaca tcgaggtcgc
      841 cccggagcac cgggggcacg gggtcgggcg cgcgttgatg gggctcgcga cggagttcgc
      901 ccgcgagcgg ggcgccgggc acctctggct ggaggtcacc aacgtcaacg caccggcgat
      961 ccacgcgtac cggcggatgg ggttcaccct ctgcggcctg g
//
