LOCUS       sequence-165977-        1018 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-3 (Chr X: 
            223616..224744).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1018)
  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 1018)
  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: pCfB3034(X-3 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..1018
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    217..250
                     /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)"
     primer_bind     879..901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     893..910
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     894..910
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     920..939
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        920..938
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa
       61 aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca
      121 acgataatac aaaatataat acaaactata agatgttatc agtatttatt atgcatttag
      181 aataaatttt gtgtcgccct tattcgactc actatagaag ttcctattct ctagaaagta
      241 taggaacttc acttcatttt atttaaattt gcggccgcgc tgaggtctta atgaggtggt
      301 tattgatcac cggaaagttt ggcattaaaa acaaacgaca gcacaaagga actttcaccc
      361 tacagcacct tctgtgtgca tgattacatg tacccgagta gcgaaaaaag ccagaaaatg
      421 tgcaagaata aaggtatctc aatgaacgag ctcgagcgtc aagagacggg tataagcgaa
      481 gaagcgcaat gccgccagcc aagcaaagcc aaacgaagag atcgaccttc catggggtcg
      541 caacttttcc cggtgacctc tacatgtgtg attgttagtt cagcgtaaca gagtttcaac
      601 aacatgcata tacgagtgtg ccttcgtgcc tgattttggt tgattatccg gcttccaacc
      661 tcgttaaatt ttcataaagt gcatatcgtg taatgtaaaa agtaaaaaga taaaaaagac
      721 tcatatgaaa tcttgctgca tggagacaag tgcactcgaa ttctttactt atattttcgc
      781 tcttgagctc gtccttttac tagcatatca atatccgttt cattgaaaag tggattaatg
      841 cctcagcact agtcctgcag gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      901 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggtc gcgatcttcg
      961 agcgtcccaa aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcg
//
