LOCUS       sequence-138742-        1024 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 1024)
  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 1024)
  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: pMK285 (Stag-3FLAG-Bsr)
FEATURES             Location/Qualifiers
     source          1..1024
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(116..149)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    180..301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(191..210)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     245..264
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(433..498)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(511..555)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(587..606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(588..606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(616..633)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(616..632)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(625..647)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(910..931)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggggctgcta aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc tacccggtag
       61 aatttcgagg tcgagatcct aagcttggct ggacgtaaac tcctcttcag acctaataac
      121 ttcgtatagc atacattata cgaagttata ttaagggtta ttgaatatga tcgacgcgtt
      181 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      241 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      301 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
      361 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt tatctagatt
      421 acttgtacac tacttgtcat cgtcatcctt gtagtcgatg tcatgatctt tataatcacc
      481 gtcatggtct ttgtagtcag cccgggatcg gctgtccatg tgctggcgtt cgaattttgc
      541 agcggcggtt tccttggcgc ctgcaccgga tccgagctcc aattcgccct atagtgagtc
      601 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      661 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      721 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
      781 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
      841 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
      901 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
      961 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     1021 tttt
//
