LOCUS       sequence-315763-        7457 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 S2 protein scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  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 7457)
  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-RAB_EKARev
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             2220..2567
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3342..3374
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    3699..4287
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3752..3772)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4170..4181)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4330..4451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4341..4360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4395..4414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4613..4742
                     /label=AAV2 ITR
     rep_origin      complement(4979..5567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5059..5078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5738..6529,6530..6598))
                     /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     6361..6380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6599..6703)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6729..7184)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6866..6887)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7078..7097
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7256..7385
                     /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 gtaccatggt gtccaagagc gaagaagtga
     1381 tcaaggagtt catgaggttc aaggtcaggc tggaaggaag catgaatggc cacgagttcg
     1441 agattgaggg agagggtgaa ggcagacctt atgaaggaac acagactgct aaactcaaag
     1501 tgactaaagg tggtcctctc ccattcgctt gggacatcct gtctccacag atcatgtatg
     1561 gcagcaaagc atacgtcaag cacccagccg acgtgccaga ttacatgaaa ctgagcttcc
     1621 cagagggatt caaatgggaa agggtgatgc acttcgagga tggaggactc gtcacagtta
     1681 cccaagacac ttctctccaa gacggaaccc tgatctacaa ggtcaagatg agaggcacta
     1741 actttccacc cgatggaccc gttatgcaga agaagactct cggttgggac tatgcaactg
     1801 agagactgta cccagaagac ggagtgctca agggtgaact gctcggtaga ctcaagctca
     1861 aggacggtgg cctgaacctg gtggagttca agacaatcta catggctaag aagccagtcc
     1921 agctccctgg ctactacttc gtggacacca aactgggcat tacatctcac aatgaagact
     1981 acaccatcgt ggagcagtac gaacgctccg aaggtaggca tcacctggga catggaactg
     2041 gttccaggtc catggctgat gaagagaagc tgccacctgg atgggagaag agaatgtctc
     2101 gcagctctgg cagagtttac tacttcaacc acatcacaaa cgcctcccag tgggaaaggc
     2161 catccggtaa ctcttcctcc ggtggcaaga acggacaagg cgaacctgct agaggaacca
     2221 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt
     2281 ccggcagtgc tggtggtagt gctggtggta gtaccagtgc tggtggtagt gctggtggta
     2341 gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg
     2401 gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggta
     2461 gtgctggtgg ttccggcagt gctggtggta gtgctggtgg tagtaccagt gctggtggta
     2521 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggttct ggtccagatg
     2581 tgcctcgcac tcctgtggac aaggctaaac tgagcttcca gtttccaggt actgctagca
     2641 gcgaagataa caacatggct accatcaagg agttcatgag attcaaagtt cgcatggagg
     2701 gcagcatgaa cggacacgag ttcgaaatcg agggtgaagg tgagggcaga ccttacgaag
     2761 gtactcagac tgcaaagctg aaggtgacca agggtggacc tctgccattt gcctgggaca
     2821 tcctctctcc acagttcatg tacggcagcg aggcatacgt caggcatcct gctgatatcc
     2881 ctgattacaa gaaactccca tttcctgagg gattcaaatg ggagagagtc atgaactttg
     2941 aagacggtgg tctggtcact gtgacacagg actcttctct gcaggacgga accctcatct
     3001 gcaaagttaa gatgagagga acaaactttc ctccagacgg acccgtgatg cagaagaaga
     3061 ctatgggctg ggaggcttct acagagatgc tgtaccctga ggacggagtg ctgaagggtc
     3121 attcctacca agctctcaag ctgaaggacg gtggccacta tctcgtggag ttcgagacta
     3181 tctacatggc aaagaaaccc gtgcaactgc ctggtgacta ctgtgttgac acaaagctgg
     3241 acattacaag ccacaacgag gattacacca tcgttgagca atacggaagg tctgagggca
     3301 ggcataggct cggcatggat gagaggtaca agggatccgg ttacactgac attgagatga
     3361 acagactcgg taagggaggt tctggaggtt ctggtggtac tggtggttct ggaggatctg
     3421 gtggtacagg aggtagcgga ggttctggtg gaaccggaag cagagtgtac gagtctgaga
     3481 aactggctag agaggcagac aagctggctc agaagagcga ggatatggct agagaggcag
     3541 ataagcaggc caggagggct gaggaaagac ctgacagaga agagattgct aggctcgctg
     3601 ccatcattgc taggatggtg gctctcaact ccagaatcgc aatgctcatg gcacgcatga
     3661 tcatgctgaa ctctcaagag agctaaaagc ttatcgataa tcaacctctg gattacaaaa
     3721 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3781 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3841 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3901 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3961 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     4021 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     4081 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     4141 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     4201 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     4261 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag
     4321 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     4381 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4441 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     4501 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc
     4561 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     4621 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4681 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4741 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6721 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6781 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6841 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6901 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6961 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     7021 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     7081 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     7141 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     7201 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg
     7261 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     7321 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     7381 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
     7441 gatcgtaggt taattaa
//
