LOCUS       sequence-165972-         969 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-2 (Chr X: 
            194944..195980).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 969)
  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 969)
  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: pCfB2899(X-2 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..969
                     /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     828..850
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     842..859
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     843..859
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     869..888
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        869..887
                     /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 atctcgccaa
      301 ggcattacca tcccatgtaa gaacggaata aaacagcatt cgaaggttag gtgtggcaca
      361 aagccgataa gaaaaagaaa agtagtgagg acaggcttaa ttgagctaat taaggcgaaa
      421 aaaggaaaag aactcgatag aaaaacttat tcatgaagga gacgtaaagt caggcaaggc
      481 tgcggcatca tgtctacttg tcataactca atttgcctat ttcttacggc ttctcataaa
      541 acgtcccaca ctattcaggg gaaggctaca cggaaacccc aataaaggaa acgaagaagt
      601 gaccttagcc ttcgtcatcc gcgcccacta ctctcttcct agtcgcccgg ttgttgaaag
      661 tttaattgtg ggttttcttt cacgacacac ctcactgaaa aaagaaacga gcggaggaat
      721 agtatgataa atcttcagca tagatgggta acgggatccc tctgtgaggc cgattatgca
      781 ggattaatgc ctcagcacta gtcctgcagg ggtaacgcca gggttttccc agtcacgacg
      841 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggacg
      901 cgatcttcga gcgtcccaaa accttctcaa gcaaggtttt cagtataatg ttacatgcgt
      961 acacgcgtc
//
