LOCUS       sequence-311261-        2241 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains GFP-based fluorescent reporter GCaMP6f (calcium 
            indicator), Xpress tag, and S1 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2241)
  AUTHORS   Linghu C, Johnson SL, Valdes PA, Shemesh OA, Park WM, Park D, 
            Piatkevich KD, Wassie AT, Liu Y, An B, Barnes SA, Celiker OT, Yao 
            CC, Yu CJ, Wang R, Adamala KP, Bear MF, Keating AE, Boyden ES
  TITLE     Spatial Multiplexing of Fluorescent Reporters for Imaging Signaling 
            Network Dynamics.
  JOURNAL   Cell. 2020 Nov 17. pii: S0092-8674(20)31399-4. doi: 
            10.1016/j.cell.2020.10.035.
  PUBMED    33232692
REFERENCE   2  (bases 1 to 2241)
  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: "Cell. 2020 
            Nov 17. pii: S0092-8674(20)31399-4. doi: 
            10.1016/j.cell.2020.10.035."
COMMENT     Sequence Label: pAAV-UBC-S1-GCaMP6f
FEATURES             Location/Qualifiers
     source          1..2241
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(238..246,247..264,265..267,268..300,301..303,304..327,
                     328..354,355..411,412..417,418..687,688..705,706..1140,
                     1141..1146,1147..1587)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      334..343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     624..645
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(751..772)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1012..1031)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1806..1820
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
ORIGIN
        1 atggcagaag cagaaagcgc actggagtat gcacaacaag cactggagaa ggctcagctg
       61 gcactccaag ctgcaagaca ggctctcaag gctggaggat ccggtggaac cggtggaagt
      121 ggagggacag gaggttcagg tggtactggt ggctctggag gaacaggagg ctccggaggt
      181 acaggagggt caggagggac tggaggctct ggaggtactg gcggttcagg aggtaccatg
      241 ggttctcatc atcatcatca tcatggtatg gcttccatga ctggtggaca gcaaatgggt
      301 cgggatctgt acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt
      361 aagtggaata agacaggtca cgcagtcaga gctataggtc ggctgagttc actcgagaac
      421 gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac
      481 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
      541 gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa
      601 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
      661 actctcggca tggacgagct gtacaagggc ggaaccggag ggagcatggt gagcaagggc
      721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      781 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
      841 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      901 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      961 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
     1021 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1141 ctgccggacc aactgactga agagcagatc gcagaattta aagaggagtt ctccctattt
     1201 gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg
     1261 gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt
     1321 gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacagg
     1381 gacacggaag aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac
     1441 atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa
     1501 gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa
     1561 gagtttgtac aaatgatgac agcgaaggga ggttcaggtg gaaccggtgg aagtggaggt
     1621 accatgaaga tggaggagct gttcaagaag cataagattg tggctgtgct gagagctaac
     1681 tctgtggagg aagctaagaa gaaggctctg gctgtgttcc tgggaggcgt tcacctgatc
     1741 gagattactt tcacagtccc agatgccgat accgtgatca aggaactgtc tttcctgaaa
     1801 gaaatgggag ccatcattgg agctggcacc gtcacctccg tcgaacaatg taggaaagca
     1861 gttgagtccg gtgctgagtt catcgtctct ccacacctgg acgaggagat ttctcagttc
     1921 tgcaaagaga aaggtgtgtt ctacatgcct ggcgtcatga cacctactga actcgtcaaa
     1981 gccatgaagc tcggacatac catcctgaag ctgttcccag gagaagtggt gggaccacag
     2041 ttcgtgaaag ccatgaaagg acccttccct aatgtgaagt ttgtgcccac tggtggtgtg
     2101 aatctggata acgtgtgtga gtggttcaag gcaggcgttc tcgctgtcgg agttggttct
     2161 gcactcgtga agggtacacc tgtggaagtc gctgagaagg caaaggcttt cgttgagaag
     2221 attagaggat gtactgagta a
//
