LOCUS       sequence-98782-e         865 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expresses two green fluorescent protein variants, a short 
            fluorescence lifetime GFP variant in the nucleus and a GFP variant 
            with a long lifetime in the cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 865)
  AUTHORS   Joosen L, Hink MA, Gadella TW Jr, Goedhart J
  TITLE     Effect of fixation procedures on the fluorescence lifetimes of 
            Aequorea victoria derived fluorescent proteins.
  JOURNAL   J Microsc. 2014 Sep 1. doi: 10.1111/jmi.12168.
  PUBMED    25179491
REFERENCE   2  (bases 1 to 865)
  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: "J Microsc. 
            2014 Sep 1. doi: 10.1111/jmi.12168."
COMMENT     Sequence Label: 3xNLS-SGFP2(T65G)-T2A-SGFP2-3xNES
FEATURES             Location/Qualifiers
     source          1..865
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             92..112
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             116..136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             140..160
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      173..182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(179..181,182..184,185..697)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(224..245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(485..504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     832..853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct
       61 caagcttcga attcccacca tgggatcaga tccaaaaaag aagagaaagg tagacccgaa
      121 gaaaaagcgt aaggtcgatc ccaaaaagaa acggaaagtg ggatcaccgg tcgccaccat
      181 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      241 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      301 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      361 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
      421 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      481 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      541 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      601 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
      661 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
      721 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      781 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
      841 gctggagttc gtgaccgccg ccggg
//
