LOCUS       sequence-19363-e         764 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 764)
  AUTHORS   Okumoto S, Looger LL, Micheva KD, Reimer RJ, Smith SJ, Frommer WB
  TITLE     Detection of glutamate release from neurons by genetically encoded 
            surface-displayed FRET nanosensors.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Jun 14. 102(24):8740-5.
  PUBMED    15939876
REFERENCE   2  (bases 1 to 764)
  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: "Proc Natl 
            Acad Sci U S A. 2005 Jun 14. 102(24):8740-5."
COMMENT     Sequence Label: pRSET FLIPE-600n
FEATURES             Location/Qualifiers
     source          1..764
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             3..25
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             45..62
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             66..98
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(102..110,111..125)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(138..140,141..143,144..656)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(183..204)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(444..463)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 attttgttta actttaagaa ggagatatac atatgcgggg ttctcatcat catcatcatc
       61 atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg
      121 ataaggatcc gggccgcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      181 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      241 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      301 ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct
      361 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      421 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      481 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      541 gcaacatcct ggggcacaag ctggagtaca actacatcag ccacaacgtc tatatcaccg
      601 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
      661 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgan ggccccgngc
      721 tgctgcccga caaccactac ctgagcaccc agtccgccct gagc
//
