LOCUS       sequence-165980-         596 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree backbone vector for the addition of 1 or 2 
            genes and promoters for integration into site X-4 (Chr X: 
            236336..237310).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 596)
  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 596)
  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: pCfB3035(X-4 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..596
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    176..209
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggact
       61 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      121 ctgggccttt cgcccgggct aattatgggg tgtcgccctt attcgactct atagtgaagt
      181 tcctattctc tagaaagtat aggaacttct gaagtgggga tttaaatgcg gccgcgctga
      241 gggtttaatc ccaaagctaa gagtcccatt ttattcttct atatgtatat tttcgatact
      301 ctaaaccacc ctacaatgta gccctatact aaatctgctc aattttcagc ttctacaagt
      361 gactcgaggc cacgtggaaa gatccaacta ctccagcaca acgattcaat ataatcgatt
      421 gctccactca taagaggcaa gaacaagctt caacttttgg taagccgccg tttataaaca
      481 gggaagatgt cctttgtcaa gggaggcaca gagcatggcc aatttggcaa attgcaggtt
      541 tttctgagtg aaaaatgaaa aagcattgta gtagagtcgg ctcactgaaa aaccgg
//
