LOCUS       sequence-327060-        7275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Freeze-frame imaging of dendritic calcium signals CaMPARI2 moiety 
            with lower Ca+ affinity (Kd=199nM) (Moeyaert et al., 2018).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7275)
  AUTHORS   Perez-Alvarez A, Huhn F, Durst CD, Franzelin A, Lamothe-Molina PJ, 
            Oertner TG
  TITLE     Freeze-Frame Imaging of Dendritic Calcium Signals With TubuTag.
  JOURNAL   Front Mol Neurosci. 2021 Mar 4;14:635820. doi: 
            10.3389/fnmol.2021.635820. eCollection 2021.
  PUBMED    33762909
REFERENCE   2  (bases 1 to 7275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Mol 
            Neurosci."; date: "2021-03-4"; volume: "14:635820. doi"; pages: " 
            10.3389/fnmol.2021.635820. eCollection 2021"
COMMENT     Sequence Label: pAAV Syn CaMPARI2-human-a-Tubulin (TubuTag)
FEATURES             Location/Qualifiers
     source          1..7275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(5437..5505,5506..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
//
