LOCUS Exported 7275 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Freeze-frame imaging of dendritic calcium signals CaMPARI2 moiety with lower Ca+ affinity (Kd=199nM) (Moeyaert et al., 2018). ACCESSION . VERSION . KEYWORDS pAAV Syn CaMPARI2-human-a-Tubulin (TubuTag) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7275) AUTHORS Alvarez AP, Huhn F, Durst CD, Franzelin A, Molina P, Oertner TG TITLE Freeze-Frame Imaging of Dendritic Calcium Signals With TubuTag JOURNAL Front. Mol. Neurosci., 04 March 2021 REFERENCE 2 (bases 1 to 7275) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7275 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 27..474 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 557..574 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 581..1021 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK" misc_feature 3196..3784 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3249..3269) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3667..3678) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3787..3803) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3788..3804) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3864..4340 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4063..4082) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4380..4520 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4380..4509 /label=AAV2 ITR rep_origin 4595..5050 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4682..4701) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4892..4913 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5067..5086) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5186..5208 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5246..5264) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5332..5436 /gene="bla" /label=AmpR promoter CDS 5437..6297 /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 complement(5655..5674) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6468..7056 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6957..6976 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 7118..7258 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 7129..7258 /label=AAV2 ITR ORIGIN 1 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg 61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt 121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg 181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc 241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc 301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg 361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg 421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat 481 tcaagctgct agcatgctgc agaacgagct tgctcttaag ttggctggac ttgatattaa 541 caagactgga ggttctcatc atcaccacca ccatggatcc gaccaactga ctgaagagca 601 gatcgcagaa tttaaagagg ctttctccct atttgacaag gacggggatg ggacaataac 661 aaccaaggag ctggggacgg tgatgcggtc tctggggcag aaccccacag aagcagagct 721 gcaggacatg atcaatgaag tagatgccga cggtgacggc acaatcgact tccctgagtt 781 cctgacaatg atggcaagaa aaatgaaaga cacagacagt gaagaagaaa ttagagaagc 841 gttccgtgtg tttgataagg atggcaatgg ctacatcagt gcagcagagc ttcgccacgt 901 gatgacaaac cttggagaga agttaacaga tgaagaggtt gatgaaatga tcagggaagc 961 agacatcgat ggggatggtc aggtaaacta cgaagagttt gtagtgatga tgacagcgaa 1021 gctcgagtgc gagaaaatct atgtgcgtga tggagtgctg acgggtgata ttcatatgat 1081 cttgttgctt gaaggaaatg cccattaccg atgtgacttc agaactactt acaaagctaa 1141 ggagaagggt gtcaagttac caggcgtgca ctatgtggac cacaccattg agattttaag 1201 ccatgacaaa gattacaaca aggttaagat ctatgagtat gctgttgctc attctggatt 1261 gcctgacaat gccagacgag gcggtaccgg cggatccatg gtgagtgcga ttaagccaga 1321 catgaagatc aaactgcgta tggaaggcaa cgtaaacggg caccactttg tgatcgacgg 1381 agatggtaca ggcaagcctt atgagggaaa acagaccatg gatcttgaag tcaaagaggg 1441 cggacctctg ccttttgcct ttgatatcct gaccactgca ttccattacg gcaacagggt 1501 attcgtgaaa tatccagaca acatacaaga ctattttaag cagtcgtttc ctaaggggta 1561 ttcgtgggaa cgaagcatga ctttcgaaga cgggggcatt tgctatgcca gaagcgacat 1621 aacaatggaa ggggacactt tctataataa agttcgattt tatggtacca actttcccgc 1681 caatggtcca gttatgcaga agaagacgct gaaatggatg ccgagctgga cgcgttcatc 1741 acgtcgtaag ttcaataaga caggtcacgc actgagagct ataggtcggc tgagctcaat 1801 gtacaagtcc ggactcagat ctcgggcgcg tgagtgcatc tccatccacg ttggccaggc 1861 tggtgtccag attggcaatg cctgctggga gctctactgc ctggaacacg gcatccagcc 1921 cgatggccag atgccaagtg acaagaccat tgggggagga gatgactcct tcaacacctt 1981 cttcagtgag acgggcgctg gcaagcacgt gccccgggct gtgtttgtag acttggaacc 2041 cacagtcatt gatgaagttc gcactggcac ctaccgccag ctcttccacc ctgagcagct 2101 catcacaggc aaggaagatg ctgccaataa ctatgcccga gggcactaca ccattggcaa 2161 ggagatcatt gaccttgtgt tggaccgaat tcgcaagctg gctgaccagt gcaccggcct 2221 tcagggcttc ttggttttcc acagctttgg tgggggaact ggttctgggt tcacctccct 2281 gctcatggaa cgtctctcag ttgattatgg caagaagtcc aagctggagt tctccattta 2341 cccagcaccc caggtttcca cagctgtagt tgagccctac aactccatcc tcaccaccca 2401 caccaccctg gagcactctg attgtgcctt catggtagac aatgaggcca tctatgacat 2461 ctgtcgtaga aacctcgata tcgagcgccc aacctacact aaccttaacc gccttattag 2521 ccagattgtg tcctccatca ctgcttccct gagatttgat ggagccctga atgttgacct 2581 gacagaattc cagaccaacc tggtgcccta cccccgcatc cacttccctc tggccacata 2641 tgcccctgtc atctctgctg agaaagccta ccatgaacag ctttctgtag cagagatcac 2701 caatgcttgc tttgagccag ccaaccagat ggtgaaatgt gaccctcgcc atggtaaata 2761 catggcttgc tgcctgttgt accgtggtga cgtggttccc aaagatgtca atgctgccat 2821 tgccaccatc aaaaccaagc gcagcatcca gtttgtggat tggtgcccca ctggcttcaa 2881 ggttggcatc aactaccagc ctcccactgt ggtgcctggt ggagacctgg ccaaggtaca 2941 gagagctgtg tgcatgctga gcaacaccac agccattgct gaggcctggg ctcgcctgga 3001 ccacaagttt gacctgatgt atgccaagcg tgcctttgtt cactggtacg tgggtgaggg 3061 gatggaggaa ggcgagtttt cagaggcccg tgaagatatg gctgcccttg agaaggatta 3121 tgaggaggtt ggtgtggatt ctgttgaagg agagggtgag gaagaaggag aggaatacta 3181 atgaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt 3241 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 3301 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 3361 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 3421 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 3481 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 3541 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 3601 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 3661 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 3721 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 3781 ccgcatcgat accgtcgacc tcgagacgcg atcgcggcgc gccggcctct ctggcctcga 3841 cctcgagtct agagatgatc tacgggtggc atccctgtga cccctcccca gtgcctctcc 3901 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca 3961 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga 4021 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct 4081 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc 4141 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat 4201 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct 4261 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca 4321 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca 4381 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc 4441 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg 4501 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg 4561 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc 4621 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 4681 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 4741 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 4801 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 4861 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 4921 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 4981 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg 5041 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 5101 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc 5161 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca 5221 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc 5281 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 5341 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 5401 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 5461 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 5521 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 5581 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 5641 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 5701 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 5761 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 5821 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 5881 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 5941 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 6001 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 6061 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 6121 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 6181 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 6241 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 6301 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 6361 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 6421 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 6481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 6541 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 6601 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 6661 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 6721 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 6781 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 6841 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 6901 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 6961 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 7021 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 7081 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc 7141 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc 7201 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc 7261 ggccgcacgc gtcga //