LOCUS       sequence-165984-        1006 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-2 (Chr XI: 
            91575..92913).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1006)
  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 1006)
  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: pCfB2903(XI-2 MarkerFree)
FEATURES             Location/Qualifiers
     source          1..1006
                     /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     900..922
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     914..931
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     915..931
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     941..960
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        941..959
                     /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 tgtgggaaga
      301 ttccgctcta ccagggaaaa acaaaattat atagaaaaga aggtaagtaa taagtaatac
      361 gaaaaaaccc aaaaaaaaaa ttcaaatacc ccaaccctgt gccctttcta tttttatggt
      421 tgaaaaggtt acagaggatc ctctggagcg gtgacaacag ttgacgcaat catggtccct
      481 ggcggtccct aaggatatct cggaggaacc gaggcacgga aacggctgca cgggtacgcc
      541 agataaggat aattcagccg tcacgcatga acagaaggat gagcatttgc tcaccttcct
      601 ggacttaact tgcgtgaatg aaaagaagaa caccaatgtt agagtagggc ttcgcaccaa
      661 actttaatca agaaatctga ggagaaatga ttctaagtct agccacgaaa actgcaaagc
      721 gcaccaaaag taagaaacga caaagttttt ctagcatttt tttgtttcat caagagaaga
      781 aacacgattt atataactgt taccgctaaa acctacatct aaacttttta atatctgaaa
      841 gcgctagtcg tgtgattaat gcctcagcac tagtcctgca ggggtaacgc cagggttttc
      901 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
      961 cgaattgggt cgcgatcttc gagcgtccca aaaccttctc aagcaa
//
