LOCUS       sequence-166064-        1016 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-5 (Chr XI: 
            11779..118967).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1016)
  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 1016)
  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: pCfB3037(XI-5 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..1016
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    216..249
                     /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     792..814
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     806..823
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     807..823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     833..852
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        833..851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag aattaagaaa
       61 ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta tacatgtcaa
      121 cgataataca aaatataata caaactataa gatgttatca gtatttatta tgcatttaga
      181 ataaattttg tgtcgccctt attcgactca ctatagaagt tcctattctc tagaaagtat
      241 aggaacttca cttcatttta tttaaatttg cggccgcgct gaggtcttaa tgatcataga
      301 tccggcactt agagaaagac catagttgac caacttagat tggctattgc gcttgctatc
      361 gaaggcaacc gataacaacg gcacacacag ggccccaagt atgcacagca ccgacgatat
      421 aagagaagac aacaaccacc tgggaatttt ttccattttc ttgcgtttct tcactgtgat
      481 gcttgtgaga atattaggat ataccaacgt gcaactctga aatgtcaaaa cggtcgtgta
      541 taaataaatg atataccagt aggtgaaatc atctcatttg tctttctaaa aaagtccatt
      601 acccttaagg ttgttgtcac aacccacgga gcagttttag aggtgcaaaa ttgaaagaat
      661 ggccacattt tgagcaattc atcgacaaca agagcttatg attaacttgg gttgtgggca
      721 attgggtgta ctatgaagca gccaatagta atgcctcagc actagtcctg caggggtaac
      781 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
      841 ctcactatag ggcgaattgg gacgcgatct tcgagcgtcc caaaaccttc tcaagcaagg
      901 ttttcagtat aatgttacat gcgtacacgc gtctgtacag aaaaaaaaga aaaatttgaa
      961 atataaataa cgttcttaat actaacataa ctataaaaaa ataaataggg acctag
//
