LOCUS       sequence-165998-        1049 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 XII-1 (Chr XII: 
            795787..796720).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1049)
  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 1049)
  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: pCfB3038(XII-1 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..1049
                     /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     927..949
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     941..958
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     942..958
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     968..987
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        968..986
                     /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 atcaatcctc
      301 gcatttcagc ttccattata tttgatgact gtttctcaat ctttatgtta tcttcttaca
      361 ccgtatgtga tgatatacta ataacacgga tacttaataa acagatgatg ttacagtttc
      421 atccaatacg aacgattatc aaattattgc aacatttccc agtgaaatct tcaccaaatg
      481 ctcatacagc gtgacaataa tgagcatttt gaattttaag gaaattaatt tgtcctctat
      541 cttttgcatc cgtgcctcct ggataataac ttcaatcctt gcaccaccat tgaacttgta
      601 acagagcttt ttttttaact tcttgtcctg gtactgcata atgaatagcg ccagaagttt
      661 cacggatgtg ctactgttgg gaagagccgc gagtatggaa cttcctaagc atcgcaagtc
      721 aggggccaga tgggtaaccg gcaaagacta attagtattg atgaagttac ttaccaatat
      781 atgtcatgag aagtggcttg taaaatttat tattacagtc actcagacag agcactaaac
      841 tgaactagtt attaaggtat gtgcagttga ttcacgggaa gattaatgcc tcagcactag
      901 tcctgcaggg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
      961 gcgcgcgtaa tacgactcac tatagggcga attgggacgc gatcttcgag cgtcccaaaa
     1021 ccttctcaag caaggttttc agtataatg
//
