LOCUS       sequence-77192-e         787 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fluorescent reporter for calcium signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 787)
  AUTHORS   Mues M, Bartholomaus I, Thestrup T, Griesbeck O, Wekerle H, Kawakami
            N, Krishnamoorthy G
  TITLE     Real-time in vivo analysis of T cell activation in the central 
            nervous system using a genetically encoded calcium indicator.
  JOURNAL   Nat Med. 2013 Jun;19(6):778-83. doi: 10.1038/nm.3180. Epub 2013 May 
            12.
  PUBMED    23685843
REFERENCE   2  (bases 1 to 787)
  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"; doi: "10.1038/nm.3180"; 
            journalName: "Nat Med"; date: "2013-06"; volume: "19"; issue: "6"; 
            pages: "778-83"
COMMENT     Sequence Label: Twitch-1 pRSETB
FEATURES             Location/Qualifiers
     source          1..787
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             14..36
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             56..73
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             77..109
                     /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(113..121,122..136)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      149..158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(155..157,158..160,161..673)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     primer_bind     complement(200..221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(461..480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ctctagaaat aattttgttt aactttaaga aggagatata catatgcggg gttctcatca
       61 tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta
      121 cgacgatgac gataaggatc ccgcggccgc caccatggtg agcaagggcg aggagctgtt
      181 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      241 cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
      301 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt
      361 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      421 gcccgaaggc tacgtccagg agcgtaccat cttcttcaag gacgacggca actacaagac
      481 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      541 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca
      601 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggcccact tcaagatccg
      661 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
      721 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
      781 caaagac
//
