LOCUS       sequence-32067-e         947 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 947)
  AUTHORS   Arosio D, Ricci F, Marchetti L, Gualdani R, Albertazzi L, Beltram F
  TITLE     Simultaneous intracellular chloride and pH measurements using a 
            GFP-based sensor.
  JOURNAL   Nat Methods. 2010 Jul . 7(7):516-8.
  PUBMED    20581829
REFERENCE   2  (bases 1 to 947)
  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 
            Methods. 2010 Jul . 7(7):516-8."
COMMENT     Sequence Label: pcDNA3-PalmPalm-ClopHensor
FEATURES             Location/Qualifiers
     source          1..947
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     54..73
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        54..72
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             158..181
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             join(188..190,191..901)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(230..251)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(491..510)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     838..859
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ttagcgactc tctggctact aagagaaccc actgcttact ggcttatcga aattaatacg
       61 actcactata gggagaccca agctggctag catgctgtgc tgtatgagaa gaaccaaaca
      121 ggttgaaaag aatgatgagg accaaaagat cgctagctgg agccacccgc agttcgaaaa
      181 aggcgccgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      241 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      301 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      361 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      421 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      481 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      541 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
      601 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
      661 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
      721 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      781 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
      841 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
      901 gcgcggatcc gcgtctggtg gntggtggtt ggtctagttc cacgtgg
//
