LOCUS Exported 6290 bp ds-DNA circular SYN 12-MAY-2021 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 pAAV-UBC-S1-jRCaMP1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6290) 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 6290) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6290 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..399 /label=UbC promoter /note="human ubiquitin C promoter" regulatory 408..417 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 549..566 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 570..602 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 606..629 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 657..713 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" misc_feature 2532..3120 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2585..2605) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3003..3014) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3163..3284 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3174..3193) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3228..3247 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3446..3575 /label=AAV2 ITR primer_bind complement(3641..3658) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3812..4400) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3892..3911) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4571..5431) /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 5194..5213 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5432..5536) /gene="bla" /label=AmpR promoter rep_origin complement(5562..6017) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5699..5720) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5911..5930 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 6089..6218 /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 accatggcag 421 aagcagaaag cgcactggag tatgcacaac aagcactgga gaaggctcag ctggcactcc 481 aagctgcaag acaggctctc aaggctggag gatccggtgg aaccggtgga agtggaggga 541 caggttctca tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg 601 gtcgggatct gtacgacgat gacgataagg atctcgcaac aatggtcgac tcatcacgtc 661 gtaagtggaa taagacaggt cacgcagtca gagctatagg tcggctgagc tcagcgatca 721 actgcgaaat gatgtaccca gcggatggtg gtctgcgtgg ttacactcac atggcgctga 781 aagttgatgg cggcggtcac ctgtcctgtt ctttcgtgac cacctaccgc tccaaaaaga 841 ctgtcggcaa cattaagatg cctgccattc atagcgtcag ccaccgtctg gagcgcctgg 901 aggagagcga taacgaaatg tttgtcgtac agcgtgaaca cgcagttgcc aagtttgtgg 961 gcctgggtgg tggcggcggt accggaggga gcatgaactc cctgatcaag gagaacatgc 1021 gtatgaaagt ggttctggaa ggctccgtaa acggccacca gttcaaatgc actggtgaag 1081 gcgaaggcaa cccgtatatg ggcacccaga ctatgcgtat caaagtgatc gagggtggtc 1141 cgctgccgtt tgcgttcgac atcctggcga cgtcctttat gtatggctcc cgtaccttca 1201 tcaaatatcc gaaaggcatc ccggatttct ttaagcagtc cttcccggaa ggttttacct 1261 gggaacgtgt gacccgttac gaagacggcg gcgtaattac cgttatgcaa gacacgtctc 1321 tggaggatgg ctgcctggtg tatcacgtgc aggttcgcgg tgtgaacttc ccgagcaatg 1381 gtgctgtaat gcaaaagaaa accaaaggtt gggagcctac ggactcccaa accactgaag 1441 agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacaa 1501 taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc acagaagcag 1561 agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacaatc gacttccctg 1621 agttcctgat tatgatggca agaaaaatga aatacacaga cagtgaagaa gaaattagag 1681 aagcgttcgg cgtgtttgat aaggatggca atggctacat cagtgcagca gagcttcgcc 1741 acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa atgatcaggg 1801 aagcagacag cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag 1861 cgaagggagg ttcaggtgga accggtggaa gtggaggtac catgaagatg gaggagctgt 1921 tcaagaagca taagattgtg gctgtgctga gagctaactc tgtggaggaa gctaagaaga 1981 aggctctggc tgtgttcctg ggaggcgttc acctgatcga gattactttc acagtcccag 2041 atgccgatac cgtgatcaag gaactgtctt tcctgaaaga aatgggagcc atcattggag 2101 ctggcaccgt cacctccgtc gaacaatgta ggaaagcagt tgagtccggt gctgagttca 2161 tcgtctctcc acacctggac gaggagattt ctcagttctg caaagagaaa ggtgtgttct 2221 acatgcctgg cgtcatgaca cctactgaac tcgtcaaagc catgaagctc ggacatacca 2281 tcctgaagct gttcccagga gaagtggtgg gaccacagtt cgtgaaagcc atgaaaggac 2341 ccttccctaa tgtgaagttt gtgcccactg gtggtgtgaa tctggataac gtgtgtgagt 2401 ggttcaaggc aggcgttctc gctgtcggag ttggttctgc actcgtgaag ggtacacctg 2461 tggaagtcgc tgagaaggca aaggctttcg ttgagaagat tagaggatgt actgagtaaa 2521 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 2581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 2641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 2701 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 2761 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 2821 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 2881 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 2941 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 3001 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 3061 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 3121 atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt 3181 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 3241 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca 3301 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc 3361 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag 3421 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct 3481 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 3541 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg 3601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 3661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 3721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 3781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 3841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 3901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 3961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 4261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 4381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 4441 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 4501 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 4561 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 4621 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 4681 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 4741 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 4801 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 4861 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 4921 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 4981 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5041 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5101 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 5161 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 5221 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 5281 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 5341 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 5401 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 5461 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 5521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat 5581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 5641 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 5701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 5761 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 5821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 5881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 5941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 6001 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc 6061 ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg 6121 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg 6181 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc 6241 atgctactta tctacgtagc catgctctag gaagatcgta ggttaattaa //