LOCUS       sequence-315528-        6638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains GFP-based fluorescent reporter ExRai-CKAR (PKC 
            indicator), OLLAS tag, and S4 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6638)
  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 6638)
  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-S4-ExRaiCKAR
FEATURES             Location/Qualifiers
     source          1..6638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             2124..2165
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     misc_feature    2880..3468
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2933..2953)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3351..3362)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3511..3632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3522..3541)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3576..3595
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3794..3923
                     /label=AAV2 ITR
     rep_origin      complement(4160..4748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4240..4259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4919..5710,5711..5779))
                     /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     5542..5561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5780..5884)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5910..6365)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6047..6068)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6259..6278
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6437..6566
                     /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 accatgatta
      421 ctgtgttcgg actcaagagc aaactggcac caagaagaga gaagctcgca gaggttatct
      481 acagcagcct gcacctcgga ctggacattc ccaagggaaa gcacgctatc cgctttctgt
      541 gtctggagaa ggaggacttc tactatcctt tcgatcgctc cgacgactac actgtgattg
      601 agatcaacct catggctgga cgcagcgaag agactaagat gctcctcatc tttctgctgt
      661 tcattgcact cgaacgcaag ctgggaatca gggctcatga cgttgagatc accatcaagg
      721 agcagccagc tcattgctgg ggattcagag gacgcactgg agatagcgca cgcgacctgg
      781 attacgatat ctatgtggga ggttctggag gttctggtgg tactggtggt tctggaggat
      841 ctggtggtac aggaggtagc ggaggttctg gtggaaccgg aggtagattc agacgcttta
      901 ggactctcaa agacaaggct aaggctggag gtaccggagg atctgagctg tctgcaatca
      961 atgtgtacat caaagctgat aagcagaaga acggcatcaa ggctaacttc aagattcgcc
     1021 acaacattga ggacggaggc gtccagctcg cttaccacta ccagcagaac acacctattg
     1081 gcgacggtcc agttctcctg cctgacaatc actacctgtc tgtgcagagc aaactgagca
     1141 aagacccaaa cgagaagaga gatcacatgg tcctgctgga gtttgtgact gctgctggca
     1201 tcactctggg aatggatgaa ctgtacaagg gtggaaccgg aggtagcatg gttagcaagg
     1261 gtgaggaact ctttactggc gtggtgccta tcctcgtcga gctggacggt gatgttaacg
     1321 gacacaagtt ctccgtcagc ggagaaggtg agggtgacgc aacctacggc aaactcactc
     1381 tgaagttcat ttgtaccact ggaaagctgc ctgtgccctg gccaactctg gtgactacac
     1441 tgacttacgg agtgcagtgc ttctctcgct atcctgatca catgaagcag catgacttct
     1501 tcaagagcgc tatgccagag ggctacatcc aagagagaac tatcttcttc aaggatgacg
     1561 gaaactacaa gacaagagca gaggttaagt ttgagggaga cacactggtt aacaggattg
     1621 agctgaaagg catcgacttc aaagaagatg gaaacatcct gggacacaag ctggagtaca
     1681 attctatgca caagttctct caggagcaga tcggagagaa catcgtgtgc agagtgatct
     1741 gcacaactgg ccagatccct attagggatc tgagcgcaga catttcccaa gtcctgaagg
     1801 agaagaggtc catcaagaaa gtttggacat ttggtcgcaa ccctgcttgt gactaccacc
     1861 tgggaaacat ttccaggctg tccaacaagc acttccagat tctcctcgga gaagatggaa
     1921 acctgctcct caacgacatc tccacaaacg gaacttggct gaacggacag aaggtcgaga
     1981 agaactccaa ccagctgctg tctcaaggcg acgagattac tgttggtgtt ggcgtggaga
     2041 gcgacatcct gtccctcgtt atcttcatca acgataagtt caaacagtgc ctcgaacaga
     2101 acaaggtgga tcgcggatcc ggtagcggtt tcgcaaacga gctgggacca aggctgatgg
     2161 gtaagggagg ctccggaggt acaggagggt caggagggac tggaggctct ggaggtactg
     2221 gcggttcagg aggaaccggt atgaccgact acattagaga tggatctgct atcaaagctc
     2281 tctcctttgc catcatcctg gctgaagctg acctgagaca catcccacaa gacctgcaaa
     2341 ggctcgctgt tcgcgtgatt cacgcctgcg gtatggtcga tgtcgccaac gatctggcct
     2401 ttagcgaagg tgctggcaag gcaggtagaa acgctctgct ggctggagca cctatcctct
     2461 gcgatgctag gatggtggct gaaggaatca ccaggtccag gctgcctgct gacaatcgcg
     2521 tcatctatac actgagcgat ccttccgttc ctgagctggc caagaagatt ggaaacactc
     2581 gctctgcagc agctctcgac ctgtggctgc cacacattga aggaagcatc gtggcaattg
     2641 gcaacgcacc aactgccctg tttaggctct tcgaactcct ggacgcaggt gctcctaagc
     2701 ctgccctgat catcggtatg cctgtcggat ttgtcggtgc agctgaatcc aaggacgagc
     2761 tggcagccaa cagcagaggc gtgccctacg tgatcgtgag aggtagaaga ggtggatctg
     2821 caatgactgc tgcagcagtg aatgctctgg cctccgagag agagtaaaag cttatcgata
     2881 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2941 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3001 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3061 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3121 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3181 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3241 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3301 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3361 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3421 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3481 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3541 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3601 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3661 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3721 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3781 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3841 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3901 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3961 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4021 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4561 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4921 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4981 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5041 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5101 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5161 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5221 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5281 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5341 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5401 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5461 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5521 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5581 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5641 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5701 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5761 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5821 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5881 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     5941 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6001 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6061 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6121 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6181 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6241 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6301 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6361 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6421 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     6481 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     6541 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
     6601 tacgtagcca tgctctagga agatcgtagg ttaattaa
//
