LOCUS       sequence-166094-         935 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 935)
  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 935)
  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..935
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        55..323
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        344..418
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      423..442
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        492..759
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        780..854
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      859..878
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
ORIGIN
        1 tcttcgctat tacgccagct ggaatgcgtg cgatagggaa caaaagctgg agcttctttg
       61 aaaagataat gtatgattat gctttcactc atatttatac agaaacttga tgttttcttt
      121 cgagtatata caaggtgatt acatgtacgt ttgaagtaca actctagatt ttgtagtgcc
      181 ctcttgggct agcggtaaag gtgcgcattt tttcacaccc tacaatgttc tgttcaaaag
      241 attttggtca aacgctgtag aagtgaaagt tggtgcgcat gtttcggcgt tcgaaacttc
      301 tccgcagtga aagataaatg atcctctcga agtggtcacg tgcgttttag agctagaaat
      361 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtggt
      421 gctttttttg ttttttatgt cttcgagtca tgtaattagt taagtgcagg tagggaacaa
      481 aagctggagc ttctttgaaa agataatgta tgattatgct ttcactcata tttatacaga
      541 aacttgatgt tttctttaga gtatatacaa ggtgattaca tgtacgtttg aagtacaact
      601 ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac
      661 aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag taaagttggt gcgcatgttt
      721 cggcgttcga aacttctccg cagtgaaaga taaatgatct gagaatactg ttgtaaaacg
      781 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      841 gcaccgagtc ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta attagttaat
      901 ctgtcatagg gaacaaaagc tggagcttct ttgaa
//
