LOCUS       sequence-137806-         690 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  C-terminal tagging with Stag-3FLAG using CRIPSR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 690)
  AUTHORS   Natsume T, Kiyomitsu T, Saga Y, Kanemaki MT
  TITLE     Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron 
            Tagging with Short Homology Donors.
  JOURNAL   Cell Rep. 2016 Apr 5;15(1):210-8. doi: 10.1016/j.celrep.2016.03.001.
            Epub 2016 Mar 24.
  PUBMED    27052166
REFERENCE   2  (bases 1 to 690)
  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: 
            "10.1016/j.celrep.2016.03.001"; journalName: "Cell Rep"; date: 
            "2016-04-5- 5"; volume: "15"; issue: "1"; pages: "210-8"
COMMENT     Sequence Label: pMK283 (Stag-3FLAG-Neo)
FEATURES             Location/Qualifiers
     source          1..690
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             58..102
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             115..180
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    312..433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(349..368)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     403..422
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(577..598)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcgcgtaat acgactcact atagggcgaa ttggagctcg gatccggtgc aggcgccaag
       61 gaaaccgccg ctgcaaaatt cgaacgccag cacatggaca gccgatcccg ggctgactac
      121 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
      181 tagtgtacaa gtaatctaga taactgatca taatcagcca taccacattt gtagaggttt
      241 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
      301 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
      361 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
      421 tcaatgtatc ttaacgcgta aattgtaagc gttaanattt tgttaaaatt cgcgttaaat
      481 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
      541 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
      601 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
      661 ctacgtgaac catcacccta atcaagtttt
//
