LOCUS       sequence-166014-        1024 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            site X-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1024)
  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 1024)
  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: pCfB3020(gRNA X-2)
FEATURES             Location/Qualifiers
     source          1..1024
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(12..34)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        158..501
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
       61 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gctgcattta aatgatctgt
      121 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct
      181 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
      241 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
      301 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
      361 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
      421 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
      481 cacatccgaa cataaacaac catgggtacc actcttgacg acacggctta ccggtaccgc
      541 accagtgtcc cgggggacgc cgaggccatc gaggcactgg atgggtcctt caccaccgac
      601 accgttttcc gcgtcaccac caccggggac ggcttcaccc tgcgggaggt gccggtggac
      661 ccgcccctga ccaaggtgtt ccccgacgac gaatcggacg acgaatcgga cgacggggag
      721 gacggcgacc cggactcccg gacgttcgtc gcgtacgggg acgacggcga cctggcgggc
      781 ttcgtggtcg tctcgtactc cggctggaac cgccggctga ccgtcgagga catcgaggtc
      841 gccccggagc accgggggca cggggtcggg cgcgcgttga tggggctcgc gacggagttc
      901 gcccgcgagc ggggcgccgg gcacctctgg ctggaggtca ccaacgtcaa cgcaccggcg
      961 atccacgcgt accggcggat ggggttcacc ctctgcggcc tggacaccgc cctgtacgac
     1021 ggca
//
