LOCUS       sequence-166092-         999 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            sites X-2, XI-5, and XII-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 999)
  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 999)
  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: pCfB3053(gRNA X-2, XI-5, XII-4)
FEATURES             Location/Qualifiers
     source          1..999
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(7..29)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        153..496
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
       61 cgggagcaga caagcccgtc agggcgcgtc agcgggctgc atttaaatga tctgtttagc
      121 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
      181 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
      241 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
      301 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
      361 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
      421 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
      481 ccgaacataa acaaccatgg gtaccactct tgacgacacg gcttaccggt accgcaccag
      541 tgtcccgggg gacgccgagg ccatcgaggc actggatggg tccttcacca ccgacaccgt
      601 tttccgcgtc accaccaccg gggacggctt caccctgcgg gaggtgccgg tggacccgcc
      661 cctgaccaag gtgttccccg acgacgaatc ggacgacgaa tcggacgacg gggaggacgg
      721 cgacccggac tcccggacgt tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt
      781 ggtcgtctcg tactccggct ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc
      841 ggagcaccgg gggcacgggg tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg
      901 cgagcggggc gccgggcacc tctggctgga ggtcaccaac gtcaacgcac cggcgatcca
      961 cgcgtaccgg cggatggggt tcaccctctg cggcctgga
//
