LOCUS       sequence-166093-         983 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 983)
  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 983)
  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..983
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     208..225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(327..346)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(351..425)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(446..714)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     terminator      complement(764..783)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(788..862)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
       61 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      121 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      181 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      241 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggaatgca
      301 cgcgattaac taattacatg actcgaagac ataaaaaaca aaaaaagcac caccgactcg
      361 gtgccacttt ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct
      421 aaaacgttta ctcaattctt gaagcgatca tttatctttc actgcggaga agtttcgaac
      481 gccgaaacat gcgcaccaac tttcacttct acagcgtttg accaaaatct tttgaacaga
      541 acattgtagg gtgtgaaaaa atgcgcacat ttaccgctag cccaagaggg cactacaaaa
      601 tctagagttg tacttcaaac gtacatgtaa tcaccttgta tatactcgaa agaaaacatc
      661 aagtttctgt ataaatatga gtgaaagcat aatcatacat tatcttttca aagaagctcc
      721 agcttttgtt ccctatgaca gattaactaa ttacatgact cgaagacata aaaaacaaaa
      781 aaagcaccac cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac
      841 ttgctatttc tagctctaaa acgttttaca acagtattct cagatcattt atctttcact
      901 gcggagaagt ttcgaacgcc gaaacatgcg caccaacttt acttctacag cgtttgacca
      961 aaatcttttg aacagaacat tgt
//
