LOCUS       sequence-202558-        2306 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 2306)
  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 2306)
  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-2C pRSETB
FEATURES             Location/Qualifiers
     source          1..2306
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /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(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      186..195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(192..194,195..197,198..710)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             1122..1856
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVRG
                     EGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYIMADKQKNGIKANFKIRHNIE"
     primer_bind     complement(1925..1943)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1939..1986
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatcccg
      181 cggccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      241 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      301 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      361 cctggcccac cctcgtgacc accctgtcct ggggcgtgca gtgcttcgcc cgctaccccg
      421 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      481 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      541 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      601 tcctggggca caagctggag tacaactact ttagcgacaa cgtctatatc accgccgaca
      661 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
      721 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      781 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
      841 atcacatggt cctgctggag ttcgtgaccg ccgcccgcat gcaagttgcc gacgccagcg
      901 aagaggaact gagcgagtgc ttcagaatct tcgacttcga cggcaacggc ttcatcgaca
      961 gagaggagtt tggcgacatc atcagactga ccggcgagca gctgaccgac gaggacgtgg
     1021 acgagatctt cggcgactcc gacaccgaca agaacggcag aatcgatttc gacgagttcc
     1081 tgaagatggt ggaaaacgtg cagcccatct accccgagct catgggcggc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1261 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     1321 gcggtaccgg cggcagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1381 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg
     1441 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1501 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgatgtgc ttcgcccgct
     1561 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1621 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1681 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1741 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1801 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggact
     1861 aagaattcga agcttgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1921 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1981 tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg
     2041 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     2101 cattaagcgc ggcgggtgtg gtgggttacg cgcagcgtga ccgctacact tgccagcgcc
     2161 ctagcgcccg ctccctttcg ctttcttccc ttcctttctc gccaccgttc gccggctttc
     2221 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2281 tcgaccccaa aaaacttgat tagggt
//
