LOCUS       sequence-77028-e         875 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate entry vector: Position 2, HA tagged nls-EGFP with start 
            and stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 875)
  AUTHORS   Kirchmaier S, Lust K, Wittbrodt J
  TITLE     Golden GATEway cloning--a combinatorial approach to generate fusion 
            and recombination constructs.
  JOURNAL   PLoS One. 2013 Oct 7;8(10):e76117. doi: 
            10.1371/journal.pone.0076117.
  PUBMED    24116091
REFERENCE   2  (bases 1 to 875)
  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: "PLoS One.";
            date: "2013-10-7"; volume: "8(10):e76117. doi"; pages: " 
            10.1371/journal.pone.0076117"
COMMENT     Sequence Label: pGGEV_2_+nls-eGFP-HA-_+1_BK+
FEATURES             Location/Qualifiers
     source          1..875
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(48..74)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(75..785,786..788,789..791))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(75..785,786..788,789..791))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(117..138)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     466..485
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     725..746
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(837..857)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      860..869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ccttacttag ttacccggga ggatgagggt ctcaaggtgg taccctaagc gtaatctgga
       61 acatcgtatg ggtacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
      121 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg
      181 gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
      241 tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc
      301 accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg
      361 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
      421 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
      481 tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
      541 aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg
      601 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg
      661 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
      721 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
      781 ttgctcacca ttccgccacc ttctagaggc tcgagaggcc ttgaattcga tcgatttacc
      841 ttacgcttct tctttggagc catggcggcg gatcc
//
