LOCUS       sequence-315524-        7742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains GFP-based fluorescent reporter cAMPr (cAMP 
            indicator), HA tag, and S2 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7742)
  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 7742)
  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-S2-cAMPr
FEATURES             Location/Qualifiers
     source          1..7742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2621..2642
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3633..3659
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3984..4572
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4037..4057)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4455..4466)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4615..4736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4626..4645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4680..4699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4898..5027
                     /label=AAV2 ITR
     rep_origin      complement(5264..5852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5344..5363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6023..6814,6815..6883))
                     /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     6646..6665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6884..6988)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7014..7469)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7151..7172)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7363..7382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7541..7670
                     /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 accatgaagc
      421 ccaccacaat cagcctgctg cagaagtaca agcaggagaa gaagcggttc gccaccatca
      481 cagcttacga ctactccttc gccaagctgt ttgctgatga gggactgaac gtgatgctgg
      541 tgggagactc tctgggcatg accgtgcagg gacacgacag caccctgcca gtgacagtgg
      601 ccgatatcgc ttaccacaca gccgctgtga ggagaggagc ccctaactgc ctgctgctgg
      661 ctgatctgcc cttcatggcc tacgctaccc ctgagcaggc ctttgagaac gccgctacag
      721 tgatgagggc cggcgctaac atggtgaaga tcgagggcgg agagtggctg gtggagaccg
      781 tgcagatgct gacagagaga gccgtgccag tgtgcggaca cctgggactg accccacagt
      841 ctgtgaacat ctttggcgga tacaaggtgc agggaagggg actgctggct ggactgcagc
      901 tgctgtccga cgctctggct ctggaggctg ctggagctca gctgctggtg ctggagtgcg
      961 tgcctgtgga gctggctaag cgcatcacag aggccctggc tatcccagtg atcggaatcg
     1021 gagctggaaa cgtgaccgac ggacagatcc tggtcatgca cgatgctttc ggcatcacag
     1081 gcggacacat cccaaagttc gccaagaact ttctggctga gaccggcgat atcagggctg
     1141 ctgtgcgcca gtacatggct gaggtggagt ccggagtgta cccaggagag gagcacagct
     1201 ttcactctag aggaggttct ggtggaaccg gtggaagtgg agggacagga ggttcaggtg
     1261 gtactggtgg ctctggagga acaggaggct ccggaggtac aggagggtca ggagggactg
     1321 gaggctctgg aggtactggc ggttcaggag gtaccatggg caacgccgcc gccgccaaga
     1381 agggcagcga gcaggagagc gtgaaagagt tcctagccaa agccaaggaa gatttcctga
     1441 aaaaatggga gaccccttct cagaatacag cccagttgga tcagtttgat agaatcaaga
     1501 cccttggcac cggctccttt gggcgagtga tgctggtgaa gcacaaggag agtgggaacc
     1561 actacgccat gaagatctta gacaagcaga aggtggtgaa gctaaagcag atcgagcaca
     1621 ctctgaatga gaagcgcatc ctgcaggccg tcaacttccc gttcctggtc aaacttgaat
     1681 tctccttcaa ggacaactca aacctgtaca tggtcatgga gtatgtagct ggtggcgaga
     1741 tgttctccca cctacggcgg attggaaggt tcagcgagcc ccatgcccgt ttctacgcgg
     1801 cgcagatcgt cctgaccttt gagtatctgc actccctgga cctcatctac cgggacctga
     1861 agcccgagaa tcttctcatc gaccagcagg gctatattca ggtgacagac ttcggttttg
     1921 ccaagcgtgt gaaaggccgt acttggacct tgtgtgggac ccctgagtac ttggcccccg
     1981 agattatcct gagcaaaggc tacaacaagg ctgtggactg gtgggctctc ggagtcctca
     2041 tctacgagat ggctgctggt tacccaccct tcttcgctga ccagcctatc cagatctatg
     2101 agaaaatcgt ctctgggaag gtgcggttcc catcccactt cagctctgac ttgaaggacc
     2161 tgctgcggaa ccttctgcaa gtggatctca ccaagcgctt tgggaacctc aagaacgggg
     2221 tcaatgacat caagaaccac aagtggtttg ccacgactga ctggattgcc atctatcaga
     2281 gaaaggtgga agctcccttc ataccaaagt ttaaaggccc tggggacacg agtaactttg
     2341 acgactatga ggaggaagag atccgggtct ccatcaatga gaagtgtggc aaggagttta
     2401 ctgagtttcc cgggaacgtc tatatcaagg ccgacgagca gaagaacggc atcaaggcgt
     2461 acttcaagat ccgccacaac atcgagggcg gcggcgtgca gctcgcctac cactaccagc
     2521 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc
     2581 agtccatgct ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2641 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga
     2701 gcgaatccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atccaggtcg
     2761 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg
     2821 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2881 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2941 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca
     3001 ccatcttctt caagggcgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3061 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3121 tggggcacaa gctggagtac aacaccgccc cccccgcatg caagggccgg cggcgacgcg
     3181 gggccatcag cgctgaggtc tacaccgagg aggatgccgc gtcctatgtt cggaaggtta
     3241 taccgaaaga ttataagaca atggctgctt tagctaaagc cattgaaaag aacgtactgt
     3301 tttcacatct tgatgataac gagagaagtg acatttttga cgccatgttc ccggtttcct
     3361 ttattgctgg agagactgtt attcagcagg gtgacgaagg ggataacttc tacgtgattg
     3421 accaaggaga gatggacgtc tatgtcaaca atgaatgggc aaccagtgtt ggggaaggag
     3481 ggagcttcgg ggagcttgct ctgatttacg ggactcctcg agcggccact gtcaaggcca
     3541 agacgaacgt gaaactgtgg ggcattgacc gggacagcta cagaaggatc ctcatgggaa
     3601 gcacgctgag aaagcggaag atgggatccg gttacccata cgacgttcca gactacgctg
     3661 gaggttctgg aggttctggt ggtactggtg gttctggagg atctggtggt acaggaggta
     3721 gcggaggttc tggtggaacc ggaagcagag tgtacgagtc tgagaaactg gctagagagg
     3781 cagacaagct ggctcagaag agcgaggata tggctagaga ggcagataag caggccagga
     3841 gggctgagga aagacctgac agagaagaga ttgctaggct cgctgccatc attgctagga
     3901 tggtggctct caactccaga atcgcaatgc tcatggcacg catgatcatg ctgaactctc
     3961 aagagagcta aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     4021 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4081 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4141 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4201 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4261 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4321 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4381 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4441 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4501 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4561 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat
     4621 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4681 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4741 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4801 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac
     4861 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     4921 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4981 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa
     5041 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5101 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5161 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5401 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5701 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     7021 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7081 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7141 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7201 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7261 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7321 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7381 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7441 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     7501 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctgcgcgctc gctcgctcac
     7561 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     7621 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat
     7681 gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcg taggttaatt
     7741 aa
//
