LOCUS       sequence-166013-         916 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 XII-5 (Chr XII: 
            839226..840357).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 916)
  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 916)
  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: pCfB2909(XII-5 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..916
                     /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)"
     terminator      complement(742..907)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     828..852
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
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 aatctggcgg cttgagttct caacatgttt tattttttac ttatattgct
      301 ggtagggtaa aaaaatataa ctcctaggaa taggttgtct atatgttttt gtcttgcttc
      361 tataattgta acaaacaagg aaagggaaaa tactgggtgt aaaagccatt gagtcaagtt
      421 aggtcatccc ttttatacaa aatttttcaa ttttttttcc aagattcttg tacgattaat
      481 tatttttttt ttgcgtccta cagcgtgatg aaaatttcgc ctgctgcaag atgagcggga
      541 acgggcgaaa tgtgcacgcg cacaacttac gaaacgcgga tgagtcactg acagccaccg
      601 cagaggttct gactcctact gagctctatt ggaggtggca gaaccggtac cggaggaggc
      661 cgctataacc ggtttgaatt tattgtcaca gtgtcacatc agcattaagt cctcagcgag
      721 ctcgcatgga atgcgtgcga tgagcgacct catgctatac ctgagaaagc aacctgacct
      781 acaggaaaga gttactcaag aataagaatt ttcgttttaa aacctaagag tcactttaaa
      841 atttgtatac acttattttt tttataactt atttaataat aaaaatcata aatcataaga
      901 aattcgctta tttaga
//
