LOCUS       Exported                6290 bp ds-DNA     circular SYN 08-SEP-2020
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains RFP-based fluorescent reporter jRCaMP1a (calcium 
            indicator), Xpress tag, and S1 protein scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  AUTHORS   Synthetic Neurobiology
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6290)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6290)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6290)
  AUTHORS   Ed Boyden Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 17, 2020 from SnapGene Viewer 5.1.6
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     Spatial multiplexing of fluorescentreporters for imaging signaling 
            network dynamics
FEATURES             Location/Qualifiers
     source          1..6290
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          687..2792
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     misc_feature    72..201
                     /label=R-ITR
                     /note="R-ITR"
                     /note="color: #a6acb3"
     promoter        274..673
                     /label=UbC promoter
                     /note="UbC promoter"
                     /note="human ubiquitin C promoter"
                     /note="color: #ffffff"
     CDS             687..779
                     /codon_start=1
                     /label=HexCoil-Ala
                     /label=Hotag5
                     /note="Hotag5"
                     /note="color: #993366"
                     /translation="MAEAESALEYAQQALEKAQLALQAARQALKA"
     CDS             780..815
                     /codon_start=1
                     /label=Linker12
                     /translation="GGSGGTGGSGGT"
     CDS             816..2138
                     /codon_start=1
                     /label=jRCaMP1a
                     /translation="GSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKT
                     GHAVRAIGRLSSAINCEMMYPADGGLRGYTHMALKVDGGGHLSCSFVTTYRSKKTVGNI
                     KMPAIHSVSHRLERLEESDNEMFVVQREHAVAKFVGLGGGGGTGGSMNSLIKENMRMKV
                     VLEGSVNGHQFKCTGEGEGNPYMGTQTMRIKVIEGGPLPFAFDILATSFMYGSRTFIKY
                     PKGIPDFFKQSFPEGFTWERVTRYEDGGVITVMQDTSLEDGCLVYHVQVRGVNFPSNGA
                     VMQKKTKGWEPTDSQTTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAE
                     LQDMINEVDADGDGTIDFPEFLIMMARKMKYTDSEEEIREAFGVFDKDGNGYISAAELR
                     HVMTNLGEKLTDEEVDEMIREADSDGDGQVNYEEFVQMMTAK"
     CDS             2139..2174
                     /codon_start=1
                     /label=Linker12
                     /label=GGSGGTGGSGGT
                     /note="GGSGGTGGSGGT"
                     /note="color: #993366"
                     /translation="GGSGGTGGSGGT"
     CDS             2175..2789
                     /codon_start=1
                     /product="Optimized for mouse cell expression"
                     /label=I301
                     /note="I301"
                     /note="color: #993366"
                     /translation="MKMEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIEITFTV
                     PDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVESGAEFIVSPHLDEEISQFCKEKG
                     VFYMPGVMTPTELVKAMKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDN
                     VCEWFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCTE"
     misc_feature    2805..3393
                     /label=WPRE
                     /note="WPRE"
                     /note="woodchuck hepatitis 
                     virus?posttranscriptionalregulatory element"
                     /note="color: #ffffff"
     polyA_signal    3436..3557
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3719..3848
                     /label=ITR
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
                     /note="color: #ffcc99"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(4844..5704)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="bla"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #------"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5705..5809)
                     /gene="bla"
                     /label=AmpR promoter
                     /label=bla promoter
                     /note="bla promoter"
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(5835..6290)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
      121 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      181 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      241 agccatgctc taggaagatc gtaggttaat taaggcctcc gcgccgggtt ttggcgcctc
      301 ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc
      361 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
      421 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
      481 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
      541 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
      601 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
      661 atcgtcactt ggcgactagt gccaccatgg cagaagcaga aagcgcactg gagtatgcac
      721 aacaagcact ggagaaggct cagctggcac tccaagctgc aagacaggct ctcaaggctg
      781 gaggatccgg tggaaccggt ggaagtggag ggacaggttc tcatcatcat catcatcatg
      841 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      901 aggatctcgc aacaatggtc gactcatcac gtcgtaagtg gaataagaca ggtcacgcag
      961 tcagagctat aggtcggctg agctcagcga tcaactgcga aatgatgtac ccagcggatg
     1021 gtggtctgcg tggttacact cacatggcgc tgaaagttga tggcggcggt cacctgtcct
     1081 gttctttcgt gaccacctac cgctccaaaa agactgtcgg caacattaag atgcctgcca
     1141 ttcatagcgt cagccaccgt ctggagcgcc tggaggagag cgataacgaa atgtttgtcg
     1201 tacagcgtga acacgcagtt gccaagtttg tgggcctggg tggtggcggc ggtaccggag
     1261 ggagcatgaa ctccctgatc aaggagaaca tgcgtatgaa agtggttctg gaaggctccg
     1321 taaacggcca ccagttcaaa tgcactggtg aaggcgaagg caacccgtat atgggcaccc
     1381 agactatgcg tatcaaagtg atcgagggtg gtccgctgcc gtttgcgttc gacatcctgg
     1441 cgacgtcctt tatgtatggc tcccgtacct tcatcaaata tccgaaaggc atcccggatt
     1501 tctttaagca gtccttcccg gaaggtttta cctgggaacg tgtgacccgt tacgaagacg
     1561 gcggcgtaat taccgttatg caagacacgt ctctggagga tggctgcctg gtgtatcacg
     1621 tgcaggttcg cggtgtgaac ttcccgagca atggtgctgt aatgcaaaag aaaaccaaag
     1681 gttgggagcc tacggactcc caaaccactg aagagcagat cgcagaattt aaagaggctt
     1741 tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg gggacggtga
     1801 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag
     1861 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gattatgatg gcaagaaaaa
     1921 tgaaatacac agacagtgaa gaagaaatta gagaagcgtt cggcgtgttt gataaggatg
     1981 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     2041 taacagatga agaggttgat gaaatgatca gggaagcaga cagcgatggg gatggtcagg
     2101 taaactacga agagtttgta caaatgatga cagcgaaggg aggttcaggt ggaaccggtg
     2161 gaagtggagg taccatgaag atggaggagc tgttcaagaa gcataagatt gtggctgtgc
     2221 tgagagctaa ctctgtggag gaagctaaga agaaggctct ggctgtgttc ctgggaggcg
     2281 ttcacctgat cgagattact ttcacagtcc cagatgccga taccgtgatc aaggaactgt
     2341 ctttcctgaa agaaatggga gccatcattg gagctggcac cgtcacctcc gtcgaacaat
     2401 gtaggaaagc agttgagtcc ggtgctgagt tcatcgtctc tccacacctg gacgaggaga
     2461 tttctcagtt ctgcaaagag aaaggtgtgt tctacatgcc tggcgtcatg acacctactg
     2521 aactcgtcaa agccatgaag ctcggacata ccatcctgaa gctgttccca ggagaagtgg
     2581 tgggaccaca gttcgtgaaa gccatgaaag gacccttccc taatgtgaag tttgtgccca
     2641 ctggtggtgt gaatctggat aacgtgtgtg agtggttcaa ggcaggcgtt ctcgctgtcg
     2701 gagttggttc tgcactcgtg aagggtacac ctgtggaagt cgctgagaag gcaaaggctt
     2761 tcgttgagaa gattagagga tgtactgagt aaaagcttat cgataatcaa cctctggatt
     2821 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2881 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2941 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3001 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3061 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3121 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3181 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     3241 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3301 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3361 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc
     3421 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3481 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3541 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3601 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3661 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     3721 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3781 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3841 gcgcgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4981 cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5821 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5881 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5941 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6001 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6061 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6121 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6181 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6241 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
//
