LOCUS       sequence-166020-        1013 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at 
            site X-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1013)
  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 1013)
  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: pCfB3041(gRNA X-3)
FEATURES             Location/Qualifiers
     source          1..1013
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     209..226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(328..347)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(352..426)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(447..715)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
ORIGIN
        1 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
       61 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      181 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      241 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggaatgc
      301 acgcgattaa ctaattacat gactcgaaga cataaaaaac aaaaaaagca ccaccgactc
      361 ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta tttctagctc
      421 taaaactaga aacgcggaca cattaggatc atttatcttt cactgcggag aagtttcgaa
      481 cgccgaaaca tgcgcaccaa ctttcacttk tacagcgttt gaccaaaatc ttttgaacag
      541 aacattgtag ggtgtgaaaa aatgcgcacc tttaccgcta gcccaagagg gcactacaaa
      601 atctagagtt gtacttcaaa cgtacatgta atcaccttgt atatactcga aagaaaacat
      661 caagtttctg tataaatatg agtgaaagca taatcataca ttatcttttc aaagaagctc
      721 cagcttttgt tccctatcgc acgcattcca gctggcgtaa tagcgaagag gcccgcaccg
      781 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtaggcggcc
      841 gcagctcgtt ttcgacactg gatggcggcg ttagtatcga atcgacagca gtatagcgac
      901 cagcattcac atacgattga cgcatgatat tactttctgc gcacttaact tcgcatctgg
      961 gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta acatttgatt aaa
//
