LOCUS       sequence-120722-        1026 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses 3xFLAG-dCas9 in E. coli in a tetracycline-dependent manner
            for enChIP analysis to purify specific genomic regions of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1026)
  AUTHORS   Fujita T, Yuno M, Fujii H
  TITLE     An enChIP system for the analysis of bacterial genome functions.
  JOURNAL   BMC Res Notes. 2018 Jun 14;11(1):387. doi: 
            10.1186/s13104-018-3486-3.
  PUBMED    29898790
REFERENCE   2  (bases 1 to 1026)
  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: "BMC Res 
            Notes."; date: "2018-06-14"; volume: "11(1):387. doi"; pages: " 
            10.1186/s13104-018-3486-3"
COMMENT     Sequence Label: 3xFLAG-dCas9/p-bacteria
FEATURES             Location/Qualifiers
     source          1..1026
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(50..152)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(182..808)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     713..732
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        824..879
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    860..878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             897..908
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             918..983
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
       61 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
      121 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccag atatcgacgt
      181 cttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc
      241 cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata
      301 atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt
      361 gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat
      421 tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact
      481 gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag
      541 cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag
      601 ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc
      661 gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt
      721 tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg ctgttaatca
      781 ctttactttt atctaatcta gacatcatta attcctaatt tttgttgaca ctctatcgtt
      841 gatagagtta ttttaccact ccctatcagt gatagagaaa agaattcaaa agatctaaag
      901 aggagaaagg atctatggac tacaaagacc atgacggtga ttataaagat catgacatcg
      961 attacaagga tgacgatgac aagctcatgg ataagaaata ctcaataggc ttagctatcg
     1021 gcacaa
//
