LOCUS Exported 5567 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV vector expressing CaMPARI2_L398T (Kd = 825nM), a photoconvertible fluorescent protein-based calcium integrator, in neurons. ACCESSION . VERSION . KEYWORDS pAAV_hsyn_NES-his-CaMPARI2-L398T-WPRE-SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5567) AUTHORS Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE, Oertner TG, Hope BT, Schreiter ER TITLE Improved methods for marking active neuron populations. JOURNAL Nat Commun. 2018 Oct 25;9(1):4440. doi: 10.1038/s41467-018-06935-2. PUBMED 30361563 REFERENCE 2 (bases 1 to 5567) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5567 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 9..456 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 553..570 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 577..1017 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK" CDS 1813..1836 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1855..1881 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1900..1929 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" misc_feature 1951..2539 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2004..2024) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2422..2433) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2542..2558) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2543..2559) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 2588..2709 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2599..2618) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2653..2672 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2917..2934) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3088..3676) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3168..3187) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3847..4707) /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 4470..4489 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4708..4812) /gene="bla" /label=AmpR promoter rep_origin complement(4838..5293) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4975..4996) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5187..5206 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 5365..5494 /label=AAV2 ITR ORIGIN 1 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg 61 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc 121 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca 181 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact 241 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt 301 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg 361 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg 421 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcaagctg ctagcaagga 481 tccgccacca tgctgcagaa cgagcttgct cttaagttgg ctggacttga tattaacaag 541 actggaggtt ctcatcatca ccaccaccat ggatccgacc aactgactga agagcagatc 601 gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc 661 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag 721 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg 781 acaatgatgg caagaaaaat gaaagacaca gacagtgaag aagaaattag agaagcgttc 841 cgtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg 901 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac 961 atcgatgggg atggtcaggt aaactacgaa gagtttgtag tgatgatgac agcgaagctc 1021 gagtgcgaga aaatctatgt gcgtgatgga gtgctgacgg gtgatattca tatgatcttg 1081 ttgcttgaag gaaatgccca ttaccgatgt gacttcagaa ctacttacaa agctaaggag 1141 aagggtgtca agttaccagg cgtgcactat gtggaccaca ccattgagat tttaagccat 1201 gacaaagatt acaacaaggt taagatctat gagtatgctg ttgctcattc tggattgcct 1261 gacaatgcca gacgaggcgg taccggcgga tccatggtga gtgcgattaa gccagacatg 1321 aagatcaaac tgcgtatgga aggcaacgta aacgggcacc actttgtgat cgacggagat 1381 ggtacaggca agccttatga gggaaaacag accatggatc ttgaagtcaa agagggcgga 1441 cctctgcctt ttgcctttga tatcctgacc actgcattcc attacggcaa cagggtattc 1501 gtgaaatatc cagacaacat acaagactat tttaagcagt cgtttcctaa ggggtattcg 1561 tgggaacgaa gcatgacttt cgaagacggg ggcatttgct atgccagaag cgacataaca 1621 atggaagggg acactttcta taataaagtt cgattttatg gtaccaactt tcccgccaat 1681 ggtccagtta tgcagaagaa gacgctgaaa tggatgccga gctggacgcg ttcatcacgt 1741 cgtaagttca ataagacagg tcacgcaacg agagctatag gtcggctgag ctcaggaggt 1801 tctggaggtt ctgattacaa ggacgacgat gacaagggag gttctggagg ttcttaccca 1861 tacgacgtac cagattacgc tggaggttct ggaggttctg agcagaagct gatctcagag 1921 gaggacctgc gtacgtaaaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 1981 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 2041 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 2101 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 2161 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 2221 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 2281 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 2341 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 2401 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 2461 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 2521 ctttgggccg cctccccgca tcgataccgt cgacctcgac ccgggcggcc gcttcgagca 2581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 2641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 2701 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg 2761 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct 2821 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct 2881 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgact cgctgcgctc 2941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 3001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 3061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 3121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 3181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 3241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 3301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 3361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 3421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 3481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 3541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 3601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 3661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 3721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 3781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 3841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 3901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 3961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 4021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 4081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 4141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 4201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 4261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 4321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 4381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 4441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 4501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 4561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 4621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 4681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 4741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 4801 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg 4861 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc 4921 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt 4981 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc 5041 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg 5101 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga 5161 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg 5221 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg 5281 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 5341 cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca 5401 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga 5461 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc 5521 tacttatcta cgtagccatg ctctaggaag atctctgcag agggccc //