LOCUS       sequence-315754-        7298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains GFP-based fluorescent reporter ExRai-AKAR (PKA 
            indicator), V5 tag, and S3 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  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 7298)
  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-S3-ExRaiAKAR
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             3141..3182
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3540..4128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3593..3613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4011..4022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4171..4292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4182..4201)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4236..4255
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4454..4583
                     /label=AAV2 ITR
     rep_origin      complement(4820..5408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4900..4919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5579..6370,6371..6439))
                     /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     6202..6221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6440..6544)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6570..7025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6707..6728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6919..6938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7097..7226
                     /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 accatgccat
      421 tcattactgt tggacaggag aatagcacaa gcattgatct gtactacgaa gatcatggca
      481 caggcgtgcc tgttgtgctc atccacggat tcccactcag cggacactcc tgggaaaggc
      541 agtccgctgc actcctggat gctggataca gggtgattac ctatgacagg agaggattcg
      601 gtcagagcag ccaacccacc acaggctacg actacgatac ctttgcagct gacctgaaca
      661 ccgtgctcga aaccctcgat ctgcaggatg ctgtcctggt gggattctct atgggaactg
      721 gtgaagttgc taggtacgtg agcagctacg gaacagctag aatcgctgca gtcgctttcc
      781 tggcttctct cgaacctttc ctgctgaaga cagacgacaa tcctgacggt gctgctcctc
      841 aagagttctt tgatggtatt gttgctgccg tcaaggctga caggtatgcc ttctacacag
      901 gattcttcaa cgacttctac aatctggatg agaacctggg tactcgcatt tctgaggagg
      961 ctgtcaggaa ctcttggaac actgcagcct ctggtggttt ctttgcagca gcagctgcac
     1021 ccactacctg gtacacagac ttcagggcag atatcccaag gattgacgtc ccagctctga
     1081 tcctgcatgg aacaggtgat cgcacactgc ccatcgagaa cacagctaga gtctttcaca
     1141 aagcactgcc ttccgcagaa tacgttgagg tcgaaggtgc acctcacgga ctgctctgga
     1201 cccacgctga ggaggttaac actgcactgc tcgcattcct ggctaaggct ctggaagccc
     1261 agaaacagaa gctgctgacc gaggtggaga catacgtcct ctctatcatt ccctctggac
     1321 ctctgaaagc tgagattgct cagagactgg aagacgtctt tgcaggcaag aacaccgacc
     1381 tggaagttct gatggaatgg ctgaagacta ggccaattct gagccctctc accaaaggaa
     1441 tcctcggttt cgtctttact ctgacagtgc ctagcgaaag aggtctgcaa cgcaggagat
     1501 tcgttcagaa tgctctgaat ggtaacggag atccaaacaa catggacaaa gctgtgaaac
     1561 tgtaccgcaa actcaaacgc gagattacct tccacggagc taaagaaatc tctctcagct
     1621 attctgcagg tgcactcgct tcttgcatgg gactgatcta caatagaatg ggtgcagtta
     1681 caacagaagt tgcattcggt ctggtgtgtg caacctgcga acagattgcc gacagccagc
     1741 atcgctctca tagacaactc gagtctagag gaggttctgg tggaaccggt ggaagtggag
     1801 ggacaggagg ttcaggtggt actggtggct ctggaggaac aggaggctcc ggaggtacag
     1861 gagggtcagg agggactgga ggctctggag gtactggcgg ttcaggaggt accggaggtc
     1921 tgagaagagc tacactcgtt gatggaggta ccggaggatc tgagctgtct gcaatcaatg
     1981 tgtacatcaa agctgataag cagaagaacg gcatcaaggc taacttcaag attcgccaca
     2041 acattgagga cggaggcgtc cagctcgctt accactacca gcagaacaca cctattggcg
     2101 acggtccagt tctcctgcct gacaatcact acctgtctgt gcagagcaaa ctgagcaaag
     2161 acccaaacga gaagagagat cacatggtcc tgctggagtt tgtgactgct gctggcatca
     2221 ctctgggaat ggatgaactg tacaagggtg gaaccggagg tagcatggtt agcaagggtg
     2281 aggaactctt tactggcgtg gtgcctatcc tcgtcgagct ggacggtgat gttaacggac
     2341 acaagttctc cgtcagcgga gaaggtgagg gtgacgcaac ctacggcaaa ctcactctga
     2401 agttcatttg taccactgga aagctgcctg tgccctggcc aactctggtg actacactga
     2461 cttacggagt gcagtgcttc tctcgctatc ctgatcacat gaagcagcat gacttcttca
     2521 agagcgctat gccagagggc tacatccaag agagaactat cttcttcaag gatgacggaa
     2581 actacaagac aagagcagag gttaagtttg agggagacac actggttaac aggattgagc
     2641 tgaaaggcat cgacttcaaa gaagatggaa acatcctggg acacaagctg gagtacaatt
     2701 ctatgcacaa gttctctcag gagcagatcg gagagaacat cgtgtgcaga gtgatctgca
     2761 caactggcca gatccctatt agggatctga gcgcagacat ttcccaagtc ctgaaggaga
     2821 agaggtccat caagaaagtt tggacatttg gtcgcaaccc tgcttgtgac taccacctgg
     2881 gaaacatttc caggctgtcc aacaagcact tccagattct cctcggagaa gatggaaacc
     2941 tgctcctcaa cgacatctcc acaaacggaa cttggctgaa cggacagaag gtcgagaaga
     3001 actccaacca gctgctgtct caaggcgacg agattactgt tggtgttggc gtggagagcg
     3061 acatcctgtc cctcgttatc ttcatcaacg ataagttcaa acagtgcctc gaacagaaca
     3121 aggtggatcg cggatccggt ggtaagccta tccctaaccc tctgctcggt ctcgattcta
     3181 ccggaggttc tggaggttct ggtggtactg gtggttctgg aggatctggt ggtacaggag
     3241 gtagcggagg ttctggtgga acctctgaag aactgagagc tgtcgcagac ctgcaaagac
     3301 tcaacattga actggccagg aagctgctgg aagctgtggc tagactgcag gaactcaaca
     3361 ttgatctcgt gaggaagacc tccgaactga ccgatgagaa gacaatcagg gaggagatca
     3421 gaaaggtgaa ggaagagtcc aagaggattg tggaggaagc cgaagaagaa atcagaagag
     3481 ccaaggaaga atccaggtac atcgcagatg agtccagagg aagctaaaag cttatcgata
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3961 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4081 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     4141 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     4201 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4261 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4321 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4381 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     4441 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4501 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4561 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4621 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4681 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4741 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4801 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4981 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5221 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6541 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     6601 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6661 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6721 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6781 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6841 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6901 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6961 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7021 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7081 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     7141 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     7201 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
     7261 tacgtagcca tgctctagga agatcgtagg ttaattaa
//
