LOCUS       sequence-166061-        1000 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 XI-1 (Chr XI: 
            67491..68573).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1000)
  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 1000)
  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: pCfB3036(XI-1 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..1000
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    175..208
                     /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(748..913)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     834..858
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggactc
       61 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
      121 tgggcctttc gcccgggcta attatggggt gtcgccctta ttcgactcta tagtgaagtt
      181 cctattctct agaaagtata ggaacttctg aagtggggat ttaaatgcgg ccgcgctgag
      241 ggtttaatgg aatagtgacg ttgtgatgcg gtgagttcgg cggttagggg aatggtatat
      301 gataaaaaac ggaaacgtgc ttctttaatt taattgttta atattgttgc agatatataa
      361 aaagggggaa agaaccgaag atgtaattat ttttttatcg cctcaaccta aagcaagcaa
      421 taaggtataa agatcaggac gtctcgagcg ctgatatcta aatttgaagc cacgcaagta
      481 actacgtagg tcagagggga caaggaataa cacttgacat ttttcttttt tctttttttt
      541 tctttttttt ttttttgtta atcttggctt ctgtaccgta gcctcctgta tacggtttta
      601 gatgcagtat gtttatcatc gccgagaaat ttggggggtg caaaggaata aaatccgtgc
      661 tgaaatccgt gcaccgcatc aaactttctc ggaggattct ttgcagccga ttaagtcctc
      721 agcgagctcg catggaatgc gtgcgatgag cgacctcatg ctatacctga gaaagcaacc
      781 tgacctacag gaaagagtta ctcaagaata agaattttcg ttttaaaacc taagagtcac
      841 tttaaaattt gtatacactt atttttttta taacttattt aataataaaa atcataaatc
      901 ataagaaatt cgcttattta gaagtgtcaa caacgtatct accaacggaa tgcgtgcgat
      961 cgcgtgcatt catccgctct aaccgaaaag gaaggagtta
//
