LOCUS       sequence-79671-e         865 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 865)
  AUTHORS   Thestrup T, Litzlbauer J, Bartholomaus I, Mues M, Russo L, Dana H, 
            Kovalchuk Y, Liang Y, Kalamakis G, Laukat Y, Becker S, Witte G, 
            Geiger A, Allen T, Rome LC, Chen TW, Kim DS, Garaschuk O, Griesinger
            C, Griesbeck O
  TITLE     Optimized ratiometric calcium sensors for functional in vivo imaging
            of neurons and T lymphocytes.
  JOURNAL   Nat Methods. 2014 Jan 5. doi: 10.1038/nmeth.2773.
  PUBMED    24390440
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: "Nat 
            Methods. 2014 Jan 5. doi: 10.1038/nmeth.2773."
COMMENT     Sequence Label: Twitch-3 pRSETB
FEATURES             Location/Qualifiers
     source          1..865
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             17..39
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             59..76
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             80..112
                     /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(116..124,125..139)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      152..161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(158..160,161..163,164..676)
                     /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(203..224)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(464..483)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     811..832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tccctctaga aataattttg tttaacttta agaaggagat atacatatgc ggggttctca
       61 tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct
      121 gtacgacgat gacgataagg atcccgcggc cgccaccatg gtgagcaagg gcgaggagct
      181 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      241 cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
      301 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg
      361 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      421 catgcccgaa ggctacgtcc aggagcgtac catcttcttc aaggacgacg gcaactacaa
      481 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      541 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacatcag
      601 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggccc acttcaagat
      661 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
      721 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
      781 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      841 ccgcatgcaa gatgctagcg aagaa
//
