LOCUS       sequence-138180-         972 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  C-terminal tagging with mClover using CRIPSR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 972)
  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 972)
  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: pMK278 (mClover-Hygro)
FEATURES             Location/Qualifiers
     source          1..972
                     /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             join(64..66,67..69,70..783)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(109..130)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     717..738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(938..957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cgcgcgtaat acgactcact atagggcgaa ttggagctcg gatccggtgc aggcgccgct
       61 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      121 gacggcgacg taaacggcca caagttcagc gtccgcggcg agggcgaggg cgatgccacc
      181 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      241 accctcgtga ccaccttcgg ctacggcgtg gcctgcttca gccgctaccc cgaccacatg
      301 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      361 tctttcaagg acgacggtac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      421 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      481 cacaagctgg agtacaactt caacagccac aacgtctata tcacggccga caagcagaag
      541 aacggcatca aggctaactt caagatccgc cacaacgttg aggacggcag cgtgcagctc
      601 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      661 cactacctga gccatcagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
      721 gtcctgctgg agttcgtgac cgccgccggg attacacatg gcatggacga gctgtacaag
      781 taatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
      841 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
      901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      961 aataaagcat tt
//
