LOCUS       Exported                7742 bp ds-DNA     circular SYN 08-SEP-2020
DEFINITION  Signaling reporter island (SiRI) construct for spatially multiplexed
            imaging. Contains GFP-based fluorescent reporter cAMPr (cAMP 
            indicator), HA tag, and S2 protein scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7742)
  AUTHORS   Synthetic Neurobiology
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7742)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7742)
  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     Spatial multiplexing of fluorescentreporters for imaging signaling 
            network dynamics
FEATURES             Location/Qualifiers
     source          1..7742
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          687..1478
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1485..1628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1629..3932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1629..3905
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1629..3896
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /note="color: #ffffff"
     source          1629..3896
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3897..3905
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3906..3932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3906..3932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3933..4013
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3933..4013
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3933..4013
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     misc_feature    71..260
                     /label=ITR
                     /note="ITR"
     promoter        274..672
                     /label=UBC promoter
                     /note="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             687..1478
                     /codon_start=1
                     /label=1M3U
                     /label=BP09
                     /note="BP09"
                     /translation="MKPTTISLLQKYKQEKKRFATITAYDYSFAKLFADEGLNVMLVGD
                     SLGMTVQGHDSTLPVTVADIAYHTAAVRRGAPNCLLLADLPFMAYATPEQAFENAATVM
                     RAGANMVKIEGGEWLVETVQMLTERAVPVCGHLGLTPQSVNIFGGYKVQGRGLLAGLQL
                     LSDALALEAAGAQLLVLECVPVELAKRITEALAIPVIGIGAGNVTDGQILVMHDAFGIT
                     GGHIPKFAKNFLAETGDIRAAVRQYMAEVESGVYPGEEHSFH"
     CDS             1485..1628
                     /codon_start=1
                     /label=Linker48
                     /label=48
                     /note="48"
                     /translation="GGSGGTGGSGGTGGSGGTGGSGGTGGSGGTGGSGGTGGSGGTGGS
                     GGT"
     CDS             1629..3896
                     /codon_start=1
                     /label=cAMPr
                     /note="cAMPr"
                     /note="color: #993366"
                     /translation="MGNAAAAKKGSEQESVKEFLAKAKEDFLKKWETPSQNTAQLDQFD
                     RIKTLGTGSFGRVMLVKHKESGNHYAMKILDKQKVVKLKQIEHTLNEKRILQAVNFPFL
                     VKLEFSFKDNSNLYMVMEYVAGGEMFSHLRRIGRFSEPHARFYAAQIVLTFEYLHSLDL
                     IYRDLKPENLLIDQQGYIQVTDFGFAKRVKGRTWTLCGTPEYLAPEIILSKGYNKAVDW
                     WALGVLIYEMAAGYPPFFADQPIQIYEKIVSGKVRFPSHFSSDLKDLLRNLLQVDLTKR
                     FGNLKNGVNDIKNHKWFATTDWIAIYQRKVEAPFIPKFKGPGDTSNFDDYEEEEIRVSI
                     NEKCGKEFTEFPGNVYIKADEQKNGIKAYFKIRHNIEGGGVQLAYHYQQNTPIGDGPVL
                     LPDNHYLSVQSMLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSESMVSKGEE
                     LFTGVVPIQVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKGDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTAPPACKGRRRRGAISAEVYTEEDAASYVRKVIPKDYKT
                     MAALAKAIEKNVLFSHLDDNERSDIFDAMFPVSFIAGETVIQQGDEGDNFYVIDQGEMD
                     VYVNNEWATSVGEGGSFGELALIYGTPRAATVKAKTNVKLWGIDRDSYRRILMGSTLRK
                     RKM"
     CDS             3906..3932
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="HA"
                     /translation="YPYDVPDYA"
     CDS             3933..4013
                     /codon_start=1
                     /label=Linker27
                     /label=27
                     /translation="GGSGGSGGTGGSGGSGGTGGSGGSGGT"
     CDS             4014..4241
                     /codon_start=1
                     /label=2L8HC4_15
                     /label=Tetra2
                     /translation="GSRVYESEKLAREADKLAQKSEDMAREADKQARRAEERPDREEIA
                     RLAAIIARMVALNSRIAMLMARMIMLNSQES"
     misc_feature    4257..4845
                     /label=WPRE
                     /note="WPRE"
                     /note="woodchuck hepatitis virus 
                     posttranscriptionalregulatory element"
     polyA_signal    4888..5009
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    5112..5301
                     /label=ITR
                     /note="ITR"
     rep_origin      complement(5537..6125)
                     /direction=LEFT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6296..7156)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="bla"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7157..7261)
                     /gene="bla"
                     /label=AmpR promoter
                     /label=bla promoter
                     /note="bla promoter"
                     /note="AmpR promoter"
     rep_origin      complement(7287..7742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
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 gccaccatga agcccaccac aatcagcctg ctgcagaagt
      721 acaagcagga gaagaagcgg ttcgccacca tcacagctta cgactactcc ttcgccaagc
      781 tgtttgctga tgagggactg aacgtgatgc tggtgggaga ctctctgggc atgaccgtgc
      841 agggacacga cagcaccctg ccagtgacag tggccgatat cgcttaccac acagccgctg
      901 tgaggagagg agcccctaac tgcctgctgc tggctgatct gcccttcatg gcctacgcta
      961 cccctgagca ggcctttgag aacgccgcta cagtgatgag ggccggcgct aacatggtga
     1021 agatcgaggg cggagagtgg ctggtggaga ccgtgcagat gctgacagag agagccgtgc
     1081 cagtgtgcgg acacctggga ctgaccccac agtctgtgaa catctttggc ggatacaagg
     1141 tgcagggaag gggactgctg gctggactgc agctgctgtc cgacgctctg gctctggagg
     1201 ctgctggagc tcagctgctg gtgctggagt gcgtgcctgt ggagctggct aagcgcatca
     1261 cagaggccct ggctatccca gtgatcggaa tcggagctgg aaacgtgacc gacggacaga
     1321 tcctggtcat gcacgatgct ttcggcatca caggcggaca catcccaaag ttcgccaaga
     1381 actttctggc tgagaccggc gatatcaggg ctgctgtgcg ccagtacatg gctgaggtgg
     1441 agtccggagt gtacccagga gaggagcaca gctttcactc tagaggaggt tctggtggaa
     1501 ccggtggaag tggagggaca ggaggttcag gtggtactgg tggctctgga ggaacaggag
     1561 gctccggagg tacaggaggg tcaggaggga ctggaggctc tggaggtact ggcggttcag
     1621 gaggtaccat gggcaacgcc gccgccgcca agaagggcag cgagcaggag agcgtgaaag
     1681 agttcctagc caaagccaag gaagatttcc tgaaaaaatg ggagacccct tctcagaata
     1741 cagcccagtt ggatcagttt gatagaatca agacccttgg caccggctcc tttgggcgag
     1801 tgatgctggt gaagcacaag gagagtggga accactacgc catgaagatc ttagacaagc
     1861 agaaggtggt gaagctaaag cagatcgagc acactctgaa tgagaagcgc atcctgcagg
     1921 ccgtcaactt cccgttcctg gtcaaacttg aattctcctt caaggacaac tcaaacctgt
     1981 acatggtcat ggagtatgta gctggtggcg agatgttctc ccacctacgg cggattggaa
     2041 ggttcagcga gccccatgcc cgtttctacg cggcgcagat cgtcctgacc tttgagtatc
     2101 tgcactccct ggacctcatc taccgggacc tgaagcccga gaatcttctc atcgaccagc
     2161 agggctatat tcaggtgaca gacttcggtt ttgccaagcg tgtgaaaggc cgtacttgga
     2221 ccttgtgtgg gacccctgag tacttggccc ccgagattat cctgagcaaa ggctacaaca
     2281 aggctgtgga ctggtgggct ctcggagtcc tcatctacga gatggctgct ggttacccac
     2341 ccttcttcgc tgaccagcct atccagatct atgagaaaat cgtctctggg aaggtgcggt
     2401 tcccatccca cttcagctct gacttgaagg acctgctgcg gaaccttctg caagtggatc
     2461 tcaccaagcg ctttgggaac ctcaagaacg gggtcaatga catcaagaac cacaagtggt
     2521 ttgccacgac tgactggatt gccatctatc agagaaaggt ggaagctccc ttcataccaa
     2581 agtttaaagg ccctggggac acgagtaact ttgacgacta tgaggaggaa gagatccggg
     2641 tctccatcaa tgagaagtgt ggcaaggagt ttactgagtt tcccgggaac gtctatatca
     2701 aggccgacga gcagaagaac ggcatcaagg cgtacttcaa gatccgccac aacatcgagg
     2761 gcggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg
     2821 tgctgctgcc cgacaaccac tacctgagcg tgcagtccat gctttcgaaa gaccccaacg
     2881 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2941 tggacgagct gtacaagggc ggtaccggag ggagcgaatc catggtgagc aagggcgagg
     3001 agctgttcac cggggtggtg cccatccagg tcgagctgga cggcgacgta aacggccaca
     3061 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     3121 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3181 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3241 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaagggc gacggcaact
     3301 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3361 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacaccg
     3421 ccccccccgc atgcaagggc cggcggcgac gcggggccat cagcgctgag gtctacaccg
     3481 aggaggatgc cgcgtcctat gttcggaagg ttataccgaa agattataag acaatggctg
     3541 ctttagctaa agccattgaa aagaacgtac tgttttcaca tcttgatgat aacgagagaa
     3601 gtgacatttt tgacgccatg ttcccggttt cctttattgc tggagagact gttattcagc
     3661 agggtgacga aggggataac ttctacgtga ttgaccaagg agagatggac gtctatgtca
     3721 acaatgaatg ggcaaccagt gttggggaag gagggagctt cggggagctt gctctgattt
     3781 acgggactcc tcgagcggcc actgtcaagg ccaagacgaa cgtgaaactg tggggcattg
     3841 accgggacag ctacagaagg atcctcatgg gaagcacgct gagaaagcgg aagatgggat
     3901 ccggttaccc atacgacgtt ccagactacg ctggaggttc tggaggttct ggtggtactg
     3961 gtggttctgg aggatctggt ggtacaggag gtagcggagg ttctggtgga accggaagca
     4021 gagtgtacga gtctgagaaa ctggctagag aggcagacaa gctggctcag aagagcgagg
     4081 atatggctag agaggcagat aagcaggcca ggagggctga ggaaagacct gacagagaag
     4141 agattgctag gctcgctgcc atcattgcta ggatggtggc tctcaactcc agaatcgcaa
     4201 tgctcatggc acgcatgatc atgctgaact ctcaagagag ctaaaagctt atcgataatc
     4261 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4321 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4381 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4441 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4501 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4561 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4621 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     4681 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4741 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4801 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     4861 ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     4921 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4981 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5041 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     5101 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     5161 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     5221 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     5281 gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5941 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6421 ctgcaatgat accgcgcgac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7261 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     7321 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     7381 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     7441 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     7501 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     7561 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7621 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     7681 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7741 gt
//
