LOCUS       sequence-165994-         982 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-3 (Chr XI: 
            93378..94567).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 982)
  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 982)
  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: pCfB2904(XI-3 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..982
                     /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     874..896
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     888..905
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     889..905
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     915..934
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        915..933
                     /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 attgagaatc
      301 cggaccagca gataatgcat gcagtggtgt tttgatgtat aagcttgaca tttttttgct
      361 tttcccgtta tgagaaatgt tttttttaga ctgaaaataa aagaatggat agtattaaaa
      421 tagtaatgaa cgtaataagg ataggacagg agcgatagga aaagatagaa ttttggagct
      481 ttttttccct ctttttttga gcaggagcca atagtagtcg gaaagatgag tgcatagatt
      541 tttcgtattt tatttcgagt aaaaatatca cgctcttagg ttcaataaaa gaatatacta
      601 tttcttcaaa atgcttctct aatctatttt tttattttcc ttaataactg aaatattatt
      661 ttttgatgta aaatgagtga acaagaataa gggatttttg gtatgtttgc aagttctgtg
      721 gagttccatg tataaggtat acatattaga aaagctttac gattatatac taacatcatg
      781 tacaaaacaa tttaataatg atctgtattg ctggctcaat ccacgtaaat taatgcctca
      841 gcactagtcc tgcaggggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
      901 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggacgcgat cttcgagcgt
      961 cccaaaacct tctcaagcaa gg
//
