LOCUS Exported 7457 bp ds-DNA circular SYN 08-SEP-2020 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 Synthetic Neurobiology TITLE Direct Submission REFERENCE 2 (bases 1 to 7457) TITLE Direct Submission REFERENCE 3 (bases 1 to 7457) TITLE Direct Submission REFERENCE 4 (bases 1 to 7457) TITLE Direct Submission REFERENCE 5 (bases 1 to 7457) 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 SGRef: number: 4; type: "Journal Article" COMMENT Spatial multiplexing of fluorescentreporters for imaging signaling network dynamics FEATURES Location/Qualifiers source 1..7457 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 687..1478 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..3614 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..1628 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..1628 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..1628 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..1628 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..1628 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1485..1628 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 1629..3605 /organism="synthetic DNA construct" /mol_type="other DNA" source 1629..3605 /organism="synthetic DNA construct" /mol_type="other DNA" source 1629..3605 /organism="synthetic DNA construct" /mol_type="other DNA" source 1629..3605 /organism="synthetic DNA construct" /mol_type="other DNA" source 2487..2840 /organism="synthetic DNA construct" /mol_type="other DNA" source 3606..3614 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 3606..3614 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 3606..3614 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" /note="color: #ffffff" source 3648..3728 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" /note="color: #ffffff" source 3648..3728 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" source 3648..3728 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" misc_feature 71..260 /label=ITR /note="ITR" /note="color: #a6acb3" promoter 274..673 /label=UBC promoter /note="human ubiquitin C promoter" CDS 687..1478 /codon_start=1 /label=1M3U /label=BP09 /note="BP09" /note="color: #993366" /translation="MKPTTISLLQKYKQEKKRFATITAYDYSFAKLFADEGLNVMLVGD SLGMTVQGHDSTLPVTVADIAYHTAAVRRGAPNCLLLADLPFMAYATPEQAFENAATVM RAGANMVKIEGGEWLVETVQMLTERAVPVCGHLGLTPQSVNIFGGYKVQGRGLLAGLQL LSDALALEAAGAQLLVLECVPVELAKRITEALAIPVIGIGAGNVTDGQILVMHDAFGIT GGHIPKFAKNFLAETGDIRAAVRQYMAEVESGVYPGEEHSFH" CDS 1485..1628 /codon_start=1 /label=Linker48 /label=48 /note="48" /note="color: #993366" /translation="GGSGGTGGSGGTGGSGGTGGSGGTGGSGGTGGSGGTGGSGGTGGS GGT" CDS 1629..3605 /codon_start=1 /product="RAB-EKARev: Optimized for mouse cell expresion" /label=RAB-EKARev /translation="MVSKSEEVIKEFMRFKVRLEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADVPDYMKLSFPEGFKWERVMHFEDGG LVTVTQDTSLQDGTLIYKVKMRGTNFPPDGPVMQKKTLGWDYATERLYPEDGVLKGELL GRLKLKDGGLNLVEFKTIYMAKKPVQLPGYYFVDTKLGITSHNEDYTIVEQYERSEGRH HLGHGTGSRSMADEEKLPPGWEKRMSRSSGRVYYFNHITNASQWERPSGNSSSGGKNGQ GEPARGTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGG SGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAG GSAGGSGPDVPRTPVDKAKLSFQFPGTASSEDNNMATIKEFMRFKVRMEGSMNGHEFEI EGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSEAYVRHPADIPDYKKLPFP EGFKWERVMNFEDGGLVTVTQDSSLQDGTLICKVKMRGTNFPPDGPVMQKKTMGWEAST EMLYPEDGVLKGHSYQALKLKDGGHYLVEFETIYMAKKPVQLPGDYCVDTKLDITSHNE DYTIVEQYGRSEGRHRLGMDERYK" CDS 3615..3647 /codon_start=1 /product="epitope tag from vesicular stomatitis virus G protein" /label=VSV-G /translation="YTDIEMNRLGK" CDS 3648..3728 /codon_start=1 /label=Linker27 /label=27 /note="27" /note="color: #993366" /translation="GGSGGSGGTGGSGGSGGTGGSGGSGGT" CDS 3729..3956 /codon_start=1 /label=2L8HC4_15 /label=Tetra2 /note="Tetra2" /note="color: #993366" /translation="GSRVYESEKLAREADKLAQKSEDMAREADKQARRAEERPDREEIA RLAAIIARMVALNSRIAMLMARMIMLNSQES" misc_feature 3972..4560 /label=WPRE /note="WPRE" /note="woodchuck hepatitis virus posttranscriptionalregulatory element" /note="color: #a6acb3" polyA_signal 4603..4724 /label=SV40 poly(A) signal /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" /note="color: #a6acb3" misc_feature 4827..5016 /label=ITR /note="ITR" /note="color: #a6acb3; direction: LEFT" rep_origin complement(5252..5840) /direction=LEFT /label=ori /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(6011..6871) /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: #993366" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6872..6976) /gene="bla" /label=AmpR promoter /label=bla promoter /note="bla promoter" /note="AmpR promoter" /note="color: #ffffff; direction: LEFT" rep_origin complement(7002..7457) /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 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 ggtgtccaag agcgaagaag tgatcaagga gttcatgagg ttcaaggtca 1681 ggctggaagg aagcatgaat ggccacgagt tcgagattga gggagagggt gaaggcagac 1741 cttatgaagg aacacagact gctaaactca aagtgactaa aggtggtcct ctcccattcg 1801 cttgggacat cctgtctcca cagatcatgt atggcagcaa agcatacgtc aagcacccag 1861 ccgacgtgcc agattacatg aaactgagct tcccagaggg attcaaatgg gaaagggtga 1921 tgcacttcga ggatggagga ctcgtcacag ttacccaaga cacttctctc caagacggaa 1981 ccctgatcta caaggtcaag atgagaggca ctaactttcc acccgatgga cccgttatgc 2041 agaagaagac tctcggttgg gactatgcaa ctgagagact gtacccagaa gacggagtgc 2101 tcaagggtga actgctcggt agactcaagc tcaaggacgg tggcctgaac ctggtggagt 2161 tcaagacaat ctacatggct aagaagccag tccagctccc tggctactac ttcgtggaca 2221 ccaaactggg cattacatct cacaatgaag actacaccat cgtggagcag tacgaacgct 2281 ccgaaggtag gcatcacctg ggacatggaa ctggttccag gtccatggct gatgaagaga 2341 agctgccacc tggatgggag aagagaatgt ctcgcagctc tggcagagtt tactacttca 2401 accacatcac aaacgcctcc cagtgggaaa ggccatccgg taactcttcc tccggtggca 2461 agaacggaca aggcgaacct gctagaggaa ccagtgctgg tggtagtgct ggtggtagtg 2521 ctggtggtag tgctggtggt agtgctggtg gttccggcag tgctggtggt agtgctggtg 2581 gtagtaccag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg 2641 ctggtggttc cggcagtgct ggtggtagtg ctggtggtag taccagtgct ggtggtagtg 2701 ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc agtgctggtg 2761 gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg 2821 ctggtggtag tgctggtggt tctggtccag atgtgcctcg cactcctgtg gacaaggcta 2881 aactgagctt ccagtttcca ggtactgcta gcagcgaaga taacaacatg gctaccatca 2941 aggagttcat gagattcaaa gttcgcatgg agggcagcat gaacggacac gagttcgaaa 3001 tcgagggtga aggtgagggc agaccttacg aaggtactca gactgcaaag ctgaaggtga 3061 ccaagggtgg acctctgcca tttgcctggg acatcctctc tccacagttc atgtacggca 3121 gcgaggcata cgtcaggcat cctgctgata tccctgatta caagaaactc ccatttcctg 3181 agggattcaa atgggagaga gtcatgaact ttgaagacgg tggtctggtc actgtgacac 3241 aggactcttc tctgcaggac ggaaccctca tctgcaaagt taagatgaga ggaacaaact 3301 ttcctccaga cggacccgtg atgcagaaga agactatggg ctgggaggct tctacagaga 3361 tgctgtaccc tgaggacgga gtgctgaagg gtcattccta ccaagctctc aagctgaagg 3421 acggtggcca ctatctcgtg gagttcgaga ctatctacat ggcaaagaaa cccgtgcaac 3481 tgcctggtga ctactgtgtt gacacaaagc tggacattac aagccacaac gaggattaca 3541 ccatcgttga gcaatacgga aggtctgagg gcaggcatag gctcggcatg gatgagaggt 3601 acaagggatc cggttacact gacattgaga tgaacagact cggtaaggga ggttctggag 3661 gttctggtgg tactggtggt tctggaggat ctggtggtac aggaggtagc ggaggttctg 3721 gtggaaccgg aagcagagtg tacgagtctg agaaactggc tagagaggca gacaagctgg 3781 ctcagaagag cgaggatatg gctagagagg cagataagca ggccaggagg gctgaggaaa 3841 gacctgacag agaagagatt gctaggctcg ctgccatcat tgctaggatg gtggctctca 3901 actccagaat cgcaatgctc atggcacgca tgatcatgct gaactctcaa gagagctaaa 3961 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 4021 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 4081 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 4141 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 4201 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 4261 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 4321 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 4381 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 4441 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 4501 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 4561 atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt 4621 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 4681 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca 4741 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc 4801 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag 4861 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct 4921 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 4981 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg 5041 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 5101 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 5161 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 5221 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5281 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5341 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5401 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 5461 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5521 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5581 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5641 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5701 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5761 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5821 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5881 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5941 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 6001 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6061 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6121 catctggccc cagtgctgca atgataccgc gcgacccacg ctcaccggct ccagatttat 6181 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6241 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6301 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6361 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6421 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6481 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6541 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6601 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6661 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6721 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6781 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6841 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6901 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6961 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat 7021 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 7081 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 7141 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 7201 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 7261 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 7321 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 7381 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 7441 gccgctacag ggcgcgt //