LOCUS       sequence-165999-         936 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-4 (Chr 
            XII:830227..831248).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 936)
  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 936)
  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: pCfB3040(XII-4 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..936
                     /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     832..854
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     846..863
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     847..863
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     873..892
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        873..891
                     /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 attttctgcc
      301 gtacctggat ggtcatttct taacaaatcc ctaaactggt atgtaagtat ttggtacaca
      361 atttctggag gaatagccgc tcccaaagat tggcttgaca aggttttgga cttcaaatct
      421 ggcatttggc ttcttccgac tctgctgttt cctattgttt ctaatagggt acgtactgcc
      481 acgtatttaa tgagttgaat aagaacacag ttttagtatt gccccaacct tatgagcgct
      541 agttagatat aaaaggataa gggtagcaat ataaatacaa cttagttggt ggtaaaagta
      601 atgtctacaa cttttaagaa ttgtcggatc actacttcat cagatgagga ggggagggta
      661 caccaaattc cacctctaat tcaatgagcc atgtgaaata gcgcgggtaa catacctccg
      721 tgaggcatcc ttttatttga ctctaatggg gaattgtaca aattggcttc aagaattgag
      781 taaactatta atgcctcagc actagtcctg caggggtaac gccagggttt tcccagtcac
      841 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
      901 gacgcgatct tcgagcgtcc caaaaccttc tcaagc
//
