LOCUS       Exported                7276 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 7276)
  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 7276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7276
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             531..548
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             555..995
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     misc_feature    3170..3758
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3223..3243)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3641..3652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3761..3777)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3762..3778)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3845..4321
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4044..4063)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4361..4501
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4361..4490
                     /label=AAV2 ITR
     rep_origin      4576..5031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4663..4682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4873..4894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5048..5067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5167..5189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5227..5245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5313..5417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5418..6278
                     /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(5636..5655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6449..7037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6938..6957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7099..7228
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gaattcaagc tgctagcatg ctgcagaacg
      481 agcttgctct taagttggct ggacttgata ttaacaagac tggaggttct catcatcacc
      541 accaccatgg atccgaccaa ctgactgaag agcagatcgc agaatttaaa gaggctttct
      601 ccctatttga caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc
      661 ggtctctggg gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg
      721 ccgacggtga cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaatga
      781 aagacacaga cagtgaagaa gaaattagag aagcgttccg tgtgtttgat aaggatggca
      841 atggctacat cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa
      901 cagatgaaga ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa
      961 actacgaaga gtttgtagtg atgatgacag cgaagctcga gtgcgagaaa atctatgtgc
     1021 gtgatggagt gctgacgggt gatattcata tgatcttgtt gcttgaagga aatgcccatt
     1081 accgatgtga cttcagaact acttacaaag ctaaggagaa gggtgtcaag ttaccaggcg
     1141 tgcactatgt ggaccacacc attgagattt taagccatga caaagattac aacaaggtta
     1201 agatctatga gtatgctgtt gctcattctg gattgcctga caatgccaga cgaggcggta
     1261 ccggcggatc catggtgagt gcgattaagc cagacatgaa gatcaaactg cgtatggaag
     1321 gcaacgtaaa cgggcaccac tttgtgatcg acggagatgg tacaggcaag ccttatgagg
     1381 gaaaacagac catggatctt gaagtcaaag agggcggacc tctgcctttt gcctttgata
     1441 tcctgaccac tgcattccat tacggcaaca gggtattcgt gaaatatcca gacaacatac
     1501 aagactattt taagcagtcg tttcctaagg ggtattcgtg ggaacgaagc atgactttcg
     1561 aagacggggg catttgctat gccagaagcg acataacaat ggaaggggac actttctata
     1621 ataaagttcg attttatggt accaactttc ccgccaatgg tccagttatg cagaagaaga
     1681 cgctgaaatg gatgccgagc tggacgcgtt catcacgtcg taagttcaat aagacaggtc
     1741 acgcactgag agctataggt cggctgagct caatgtacaa gtccggactc agatctcggg
     1801 cgcgtgagtg catctccatc cacgttggcc aggctggtgt ccagattggc aatgcctgct
     1861 gggagctcta ctgcctggaa cacggcatcc agcccgatgg ccagatgcca agtgacaaga
     1921 ccattggggg aggagatgac tccttcaaca ccttcttcag tgagacgggc gctggcaagc
     1981 acgtgccccg ggctgtgttt gtagacttgg aacccacagt cattgatgaa gttcgcactg
     2041 gcacctaccg ccagctcttc caccctgagc agctcatcac aggcaaggaa gatgctgcca
     2101 ataactatgc ccgagggcac tacaccattg gcaaggagat cattgacctt gtgttggacc
     2161 gaattcgcaa gctggctgac cagtgcaccg gccttcaggg cttcttggtt ttccacagct
     2221 ttggtggggg aactggttct gggttcacct ccctgctcat ggaacgtctc tcagttgatt
     2281 atggcaagaa gtccaagctg gagttctcca tttacccagc accccaggtt tccacagctg
     2341 tagttgagcc ctacaactcc atcctcacca cccacaccac cctggagcac tctgattgtg
     2401 ccttcatggt agacaatgag gccatctatg acatctgtcg tagaaacctc gatatcgagc
     2461 gcccaaccta cactaacctt aaccgcctta ttagccagat tgtgtcctcc atcactgctt
     2521 ccctgagatt tgatggagcc ctgaatgttg acctgacaga attccagacc aacctggtgc
     2581 cctacccccg catccacttc cctctggcca catatgcccc tgtcatctct gctgagaaag
     2641 cctaccatga acagctttct gtagcagaga tcaccaatgc ttgctttgag ccagccaacc
     2701 agatggtgaa atgtgaccct cgccatggta aatacatggc ttgctgcctg ttgtaccgtg
     2761 gtgacgtggt tcccaaagat gtcaatgctg ccattgccac catcaaaacc aagcgcagca
     2821 tccagtttgt ggattggtgc cccactggct tcaaggttgg catcaactac cagcctccca
     2881 ctgtggtgcc tggtggagac ctggccaagg tacagagagc tgtgtgcatg ctgagcaaca
     2941 ccacagccat tgctgaggcc tgggctcgcc tggaccacaa gtttgacctg atgtatgcca
     3001 agcgtgcctt tgttcactgg tacgtgggtg aggggatgga ggaaggcgag ttttcagagg
     3061 cccgtgaaga tatggctgcc cttgagaagg attatgagga ggttggtgtg gattctgttg
     3121 aaggagaggg tgaggaagaa ggagaggaat actaatgaag cttatcgata atcaacctct
     3181 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3241 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3301 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3361 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3421 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3481 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3541 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3601 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3661 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3721 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgaga
     3781 cgcgatcgcg gcgcgccatt cgatggcctc tctggcctcg acctcgagtc tagagatgat
     3841 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     3901 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     3961 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     4021 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4081 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     4141 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4201 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4261 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4321 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4381 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4441 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4501 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4561 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4621 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     4681 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4741 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4801 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4861 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     4921 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     4981 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5041 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5101 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5161 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5221 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5281 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5341 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5401 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5461 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5521 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5581 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5641 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5701 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5761 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5821 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5881 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5941 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6001 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6061 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6241 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7081 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     7141 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     7201 gtggccaact ccatcactag gggttcctgc ggccgcacgc gtcgctagac tcgagctgca
     7261 gagggccctg cgtatg
//
