LOCUS       sequence-315764-        8048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains RFP-based fluorescent reporter RAB-EKARev (ERK 
            indicator), VSV-G tag, and S3 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8048)
  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 8048)
  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-RAB_EKARev
FEATURES             Location/Qualifiers
     source          1..8048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             2778..3125
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3900..3932
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    4290..4878
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4343..4363)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4761..4772)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4921..5042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4932..4951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4986..5005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5204..5333
                     /label=AAV2 ITR
     rep_origin      complement(5570..6158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5650..5669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6329..7120,7121..7189))
                     /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     6952..6971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7190..7294)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7320..7775)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7457..7478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7669..7688
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7847..7976
                     /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 accatggtgt
     1921 ccaagagcga agaagtgatc aaggagttca tgaggttcaa ggtcaggctg gaaggaagca
     1981 tgaatggcca cgagttcgag attgagggag agggtgaagg cagaccttat gaaggaacac
     2041 agactgctaa actcaaagtg actaaaggtg gtcctctccc attcgcttgg gacatcctgt
     2101 ctccacagat catgtatggc agcaaagcat acgtcaagca cccagccgac gtgccagatt
     2161 acatgaaact gagcttccca gagggattca aatgggaaag ggtgatgcac ttcgaggatg
     2221 gaggactcgt cacagttacc caagacactt ctctccaaga cggaaccctg atctacaagg
     2281 tcaagatgag aggcactaac tttccacccg atggacccgt tatgcagaag aagactctcg
     2341 gttgggacta tgcaactgag agactgtacc cagaagacgg agtgctcaag ggtgaactgc
     2401 tcggtagact caagctcaag gacggtggcc tgaacctggt ggagttcaag acaatctaca
     2461 tggctaagaa gccagtccag ctccctggct actacttcgt ggacaccaaa ctgggcatta
     2521 catctcacaa tgaagactac accatcgtgg agcagtacga acgctccgaa ggtaggcatc
     2581 acctgggaca tggaactggt tccaggtcca tggctgatga agagaagctg ccacctggat
     2641 gggagaagag aatgtctcgc agctctggca gagtttacta cttcaaccac atcacaaacg
     2701 cctcccagtg ggaaaggcca tccggtaact cttcctccgg tggcaagaac ggacaaggcg
     2761 aacctgctag aggaaccagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     2821 gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt accagtgctg
     2881 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggca
     2941 gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg
     3001 gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc tggtggtagt gctggtggta
     3061 gtaccagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     3121 gtggttctgg tccagatgtg cctcgcactc ctgtggacaa ggctaaactg agcttccagt
     3181 ttccaggtac tgctagcagc gaagataaca acatggctac catcaaggag ttcatgagat
     3241 tcaaagttcg catggagggc agcatgaacg gacacgagtt cgaaatcgag ggtgaaggtg
     3301 agggcagacc ttacgaaggt actcagactg caaagctgaa ggtgaccaag ggtggacctc
     3361 tgccatttgc ctgggacatc ctctctccac agttcatgta cggcagcgag gcatacgtca
     3421 ggcatcctgc tgatatccct gattacaaga aactcccatt tcctgaggga ttcaaatggg
     3481 agagagtcat gaactttgaa gacggtggtc tggtcactgt gacacaggac tcttctctgc
     3541 aggacggaac cctcatctgc aaagttaaga tgagaggaac aaactttcct ccagacggac
     3601 ccgtgatgca gaagaagact atgggctggg aggcttctac agagatgctg taccctgagg
     3661 acggagtgct gaagggtcat tcctaccaag ctctcaagct gaaggacggt ggccactatc
     3721 tcgtggagtt cgagactatc tacatggcaa agaaacccgt gcaactgcct ggtgactact
     3781 gtgttgacac aaagctggac attacaagcc acaacgagga ttacaccatc gttgagcaat
     3841 acggaaggtc tgagggcagg cataggctcg gcatggatga gaggtacaag ggatccggtt
     3901 acactgacat tgagatgaac agactcggta agggaggttc tggaggttct ggtggtactg
     3961 gtggttctgg aggatctggt ggtacaggag gtagcggagg ttctggtgga acctctgaag
     4021 aactgagagc tgtcgcagac ctgcaaagac tcaacattga actggccagg aagctgctgg
     4081 aagctgtggc tagactgcag gaactcaaca ttgatctcgt gaggaagacc tccgaactga
     4141 ccgatgagaa gacaatcagg gaggagatca gaaaggtgaa ggaagagtcc aagaggattg
     4201 tggaggaagc cgaagaagaa atcagaagag ccaaggaaga atccaggtac atcgcagatg
     4261 agtccagagg aagctaaaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     4321 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4381 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4441 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4501 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4561 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4621 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4681 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4741 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4801 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4861 tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga
     4921 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4981 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5041 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     5101 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat
     5161 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     5221 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     5281 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt
     5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5401 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5761 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6001 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6121 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6181 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6241 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6301 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6361 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6421 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6481 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6541 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6601 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6661 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6721 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6781 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6841 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6901 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6961 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7021 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7081 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7141 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7201 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7261 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     7321 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     7381 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     7441 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7501 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     7561 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7621 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7681 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7741 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     7801 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc gcgctcgctc
     7861 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
     7921 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
     7981 gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga agatcgtagg
     8041 ttaattaa
//
