LOCUS       sequence-166035-         985 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            site XII-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 985)
  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 985)
  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: pCfB3049(gRNA XII-4)
FEATURES             Location/Qualifiers
     source          1..985
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1..23)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        147..490
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
       61 cagacaagcc cgtcagggcg cgtcagcggg ctgcatttaa atgatctgtt tagcttgcct
      121 cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc
      181 ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc
      241 cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt
      301 acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt
      361 gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct
      421 tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac
      481 ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca ccagtgtccc
      541 gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca ccgttttccg
      601 cgtcaccacc accggggacg gcttcaccct gcgggaggtg ccggtggacc cgcccctgac
      661 caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg acggcgaccc
      721 ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct tcgtggtcgt
      781 ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg ccccggagca
      841 ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg cccgcgagcg
      901 gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga tccacgcgta
      961 ccggcggatg gggttcaccc tctgc
//
