LOCUS       sequence-315534-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains RFP-based fluorescent reporter jRCaMP1a (calcium 
            indicator), Xpress tag, and S1 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  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 6290)
  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-jRCaMP1a
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      408..417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             570..602
                     /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(606..614,615..629)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             657..713
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             2084..2098
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     misc_feature    2532..3120
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2585..2605)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3003..3014)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3163..3284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3174..3193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3228..3247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3446..3575
                     /label=AAV2 ITR
     rep_origin      complement(3812..4400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3892..3911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4571..5362,5363..5431))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5194..5213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5432..5536)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5562..6017)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5699..5720)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5911..5930
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6089..6218
                     /label=AAV2 ITR
ORIGIN
        1 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
       61 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
      121 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
      181 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
      241 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
      301 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
      361 cgcggttctt gtttgtggat cgctgtgatc gtcacttggc gactagtgcc accatggcag
      421 aagcagaaag cgcactggag tatgcacaac aagcactgga gaaggctcag ctggcactcc
      481 aagctgcaag acaggctctc aaggctggag gatccggtgg aaccggtgga agtggaggga
      541 caggttctca tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg
      601 gtcgggatct gtacgacgat gacgataagg atctcgcaac aatggtcgac tcatcacgtc
      661 gtaagtggaa taagacaggt cacgcagtca gagctatagg tcggctgagc tcagcgatca
      721 actgcgaaat gatgtaccca gcggatggtg gtctgcgtgg ttacactcac atggcgctga
      781 aagttgatgg cggcggtcac ctgtcctgtt ctttcgtgac cacctaccgc tccaaaaaga
      841 ctgtcggcaa cattaagatg cctgccattc atagcgtcag ccaccgtctg gagcgcctgg
      901 aggagagcga taacgaaatg tttgtcgtac agcgtgaaca cgcagttgcc aagtttgtgg
      961 gcctgggtgg tggcggcggt accggaggga gcatgaactc cctgatcaag gagaacatgc
     1021 gtatgaaagt ggttctggaa ggctccgtaa acggccacca gttcaaatgc actggtgaag
     1081 gcgaaggcaa cccgtatatg ggcacccaga ctatgcgtat caaagtgatc gagggtggtc
     1141 cgctgccgtt tgcgttcgac atcctggcga cgtcctttat gtatggctcc cgtaccttca
     1201 tcaaatatcc gaaaggcatc ccggatttct ttaagcagtc cttcccggaa ggttttacct
     1261 gggaacgtgt gacccgttac gaagacggcg gcgtaattac cgttatgcaa gacacgtctc
     1321 tggaggatgg ctgcctggtg tatcacgtgc aggttcgcgg tgtgaacttc ccgagcaatg
     1381 gtgctgtaat gcaaaagaaa accaaaggtt gggagcctac ggactcccaa accactgaag
     1441 agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacaa
     1501 taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc acagaagcag
     1561 agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacaatc gacttccctg
     1621 agttcctgat tatgatggca agaaaaatga aatacacaga cagtgaagaa gaaattagag
     1681 aagcgttcgg cgtgtttgat aaggatggca atggctacat cagtgcagca gagcttcgcc
     1741 acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa atgatcaggg
     1801 aagcagacag cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag
     1861 cgaagggagg ttcaggtgga accggtggaa gtggaggtac catgaagatg gaggagctgt
     1921 tcaagaagca taagattgtg gctgtgctga gagctaactc tgtggaggaa gctaagaaga
     1981 aggctctggc tgtgttcctg ggaggcgttc acctgatcga gattactttc acagtcccag
     2041 atgccgatac cgtgatcaag gaactgtctt tcctgaaaga aatgggagcc atcattggag
     2101 ctggcaccgt cacctccgtc gaacaatgta ggaaagcagt tgagtccggt gctgagttca
     2161 tcgtctctcc acacctggac gaggagattt ctcagttctg caaagagaaa ggtgtgttct
     2221 acatgcctgg cgtcatgaca cctactgaac tcgtcaaagc catgaagctc ggacatacca
     2281 tcctgaagct gttcccagga gaagtggtgg gaccacagtt cgtgaaagcc atgaaaggac
     2341 ccttccctaa tgtgaagttt gtgcccactg gtggtgtgaa tctggataac gtgtgtgagt
     2401 ggttcaaggc aggcgttctc gctgtcggag ttggttctgc actcgtgaag ggtacacctg
     2461 tggaagtcgc tgagaaggca aaggctttcg ttgagaagat tagaggatgt actgagtaaa
     2521 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2701 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2761 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2821 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2881 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2941 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3001 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3061 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3121 atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     3181 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3241 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3301 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3361 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     3421 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     3481 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3541 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg
     3601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4441 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4501 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4561 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4621 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4681 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4741 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4801 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4861 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4921 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4981 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5041 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5101 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5161 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5221 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5281 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5341 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5401 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5461 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     5581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     5641 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     5701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5761 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6001 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6061 ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg
     6121 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     6181 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc
     6241 atgctactta tctacgtagc catgctctag gaagatcgta ggttaattaa
//
