LOCUS       sequence-103417-        1061 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses 3xFLAG-dCas9 in budding yeast for enChIP analysis to 
            purify specific genomic regions of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1061)
  AUTHORS   Fujii H, Fujita T
  TITLE     An enChIP system for the analysis of genome functions in budding 
            yeast.
  JOURNAL   Biol Methods Protoc. 2022 Oct 17;7(1):bpac025. doi: 
            10.1093/biomethods/bpac025. eCollection 2022.
  PUBMED    36325175
REFERENCE   2  (bases 1 to 1061)
  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"; journalName: "Biol 
            Methods Protoc."; date: "2022-10-17"; volume: "7(1):bpac025. doi"; 
            pages: " 10.1093/biomethods/bpac025. eCollection 2022"
COMMENT     Sequence Label: 3xFLAG-dCas9/pTEF1p-CYC1t
FEATURES             Location/Qualifiers
     source          1..1061
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(311..376)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(391..407)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(391..407)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(411..809)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(827..847)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(827..845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(866..882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(866..882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(879..901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    890..906
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(914..920,921..938,939..944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    959..980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cctccagcct atggaagaaa gagtcatcca ccttagccat ctcattacta aagatctcct
       61 gcaggtagca gatccgattc tttctgcggg tatatctgcg ccgtgctgtt cttttgagcc
      121 gcgtggcttc ggccgtctcc ccggagtcga acaggagggc gccaatgagg ttcttcttta
      181 tgctgtggcg atcggtattg cccagaactt tgaatttttt gctcggcacc ttgtactcgt
      241 ccgtaatgac ggcccagccg acgctgtttg tgccgatagc gagcccaatg gagtacttct
      301 tgtccatgag cttgtcatcg tcatccttgt aatcgatgtc atgatcttta taatcaccgt
      361 catggtcttt gtagtccatt tttcccgggg gatccactag ttctagaaaa cttagattag
      421 attgctatgc tttctttcta atgagcaaga agtaaaaaaa gttgtaatag aacaagaaaa
      481 atgaaactga aacttgagaa attgaagacc gtttattaac ttaaatatca atgggaggtc
      541 atcgaaagag aaaaaaatca aaaaaaaaat tttcaagaaa aagaaacgtg ataaaaattt
      601 ttattgcctt tttcgacgaa gaaaaagaaa cgaggcggtc tcttttttct tttccaaacc
      661 tttagtacgg gtaattaacg acaccctaga ggaagaaaga ggggaaattt agtatgctgt
      721 gcttgggtgt tttgaagtgg tacggcgatg cgcggagtcc gagaaaatct ggaagagtaa
      781 aaaaggagta gaaacatttt gaagctatga gctccagctt ttgttccctt tagtgagggt
      841 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg g
//
