LOCUS       sequence-129953-         851 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses NLS-dCas9-GFP11x7-NLS in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 851)
  AUTHORS   Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert LA, 
            Ishikawa H, Leonetti MD, Marshall WF, Weissman JS, Huang B
  TITLE     Versatile protein tagging in cells with split fluorescent protein.
  JOURNAL   Nat Commun. 2016 Mar 18;7:11046. doi: 10.1038/ncomms11046.
  PUBMED    26988139
REFERENCE   2  (bases 1 to 851)
  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: "Nat 
            Commun."; date: "2016-03-18"; volume: "7:11046. doi"; pages: " 
            10.1038/ncomms11046"
COMMENT     Sequence Label: pHRm-NLS-dCas9-GFP11x7-NLS
FEATURES             Location/Qualifiers
     source          1..851
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(23..79)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(89..109)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(132..557)
                     /codon_start=1
                     /product="seven tandem copies of the 11th beta-strand of 
                     superfolder GFP"
                     /label=GFP11x7
                     /note="each GFP11 associates with GFP(1-10), consisting of 
                     the first 10 beta-strands, to reconstitute fluorescent GFP 
                     (Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDH
                     MVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSG
                     GRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGIT"
     CDS             complement(600..620)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(627..647)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(660..686)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             complement(666..692)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gctcgctcat acttcctggt gcaggtccag ggttctcctc cacgtctcca gcctgcttca
       61 gcaggctgaa gttagtagct ccgcttccca ccttgcgctt cttcttgggt ccacctccac
      121 cttgcggccg cggtgatacc ggcagcattg acatattcgt gcaggaccat atggtcccga
      181 ccgccgctac cgcctgtgat tccagcagcg ttcacgtact catggagtac catgtgatcc
      241 ctgccacctg atccgccagt aatgccagct gcgtttacgt attcgtgaag caccatatgg
      301 tctcgtccgc ccgacccacc agttataccc gccgcgttca catactcatg cagcaccatg
      361 tgatcccggc ctccggaccc tccggttatt ccggctgcat tgacatattc atgtagtacc
      421 atatggtcgc gtcctccact accgccagtg atgcctgcgg cgttaacgta ttcgtggaga
      481 accatatgat ctctacctcc agagccacct gtaatcccag cagcatttac atactcatga
      541 aggaccatgt ggtcacgcat ggatccgctg ctgccgttgc tcccactacc aatgccgtcc
      601 actttccgct ttttcttagg atcttccacc ttgcgttttt tcttggggga gccgctgccc
      661 aggctggcat aatcgggcac gtcataggga taggcgtcgc ctcccagctg agacaggtcg
      721 atccgtgtct cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc cagcacctct
      781 ttggtgctgg tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa ggcggcaggg
      841 gctcccagat t
//
