LOCUS       sequence-165979-         913 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-4 (Chr X: 
            236336..237310).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 913)
  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 913)
  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: pCfB3035(X-4 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..913
                     /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     784..806
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     798..815
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     799..815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     825..844
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        825..843
                     /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 atctggtgag
      301 gatttacggt atgatcatgc tggtaagctt cgttactcat gctttggttg gcttcttgaa
      361 agtgtctagt tcactttatg cccggttata aacataaact ttcacagggt tctctttgat
      421 cacctctttg tttaatctta tgaatccaag ggattttatc ggtcaaaggg gaaatcaatg
      481 cgaaagacag taatgatgag agaaaaactc tccgtaacca ccaagtttgg ttcagcgcga
      541 cgagattttt atcgattatc gagaaaaata cctgtatatc tacatttcta tgtcagtgat
      601 atatacttct tagataagtt atgccaccag tgcatacgct tacgcacaca cacgtattct
      661 tgtgcacacg cctgttactt cttgcagaca tcagacatac tattgtaatt caaaaaaaaa
      721 aagcgaatct tcccatgtat taatgcctca gcactagtcc tgcaggggta acgccagggt
      781 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
      841 agggcgaatt gggacgcgat cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt
      901 ataatgttac atg
//
