LOCUS       sequence-388361-        6611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hSyn1 driven expression of the calcium recorder Caprola_04 fused to 
            mEGFP for neuronal expression through AAV transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  AUTHORS   Huppertz MC, Wilhelm J, Grenier V, Schneider MW, Falt T, Porzberg N,
            Hausmann D, Hoffmann DC, Hai L, Tarnawski M, Pino G, Slanchev K, 
            Kolb I, Acuna C, Fenk LM, Baier H, Hiblot J, Johnsson K
  TITLE     Recording physiological history of cells with chemical labeling.
  JOURNAL   Science. 2024 Feb 23;383(6685):890-897. doi: 
            10.1126/science.adg0812. Epub 2024 Feb 22.
  PUBMED    38386755
REFERENCE   2  (bases 1 to 6611)
  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"; doi: 
            "10.1126/science.adg0812"; journalName: "Science"; date: 
            "2024-02-23- 23"; volume: "383"; issue: "6685"; pages: "890-897"
COMMENT     Sequence Label: pAAV2_hSyn_NES-Caprola_04-mEGFP_WRPE-SV40
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   43..172
                     /label=AAV2 ITR
     promoter        254..701
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             838..1278
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             1399..1455
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKFNKTGHAVRAIGRLSS"
     CDS             join(2359..2361,2362..2364,2365..3078)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2404..2425)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2665..2684)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3012..3033
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3091..3679
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3144..3164)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3562..3573)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3682..3698)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3683..3699)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3728..3849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3739..3758)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3793..3812
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4011..4140
                     /label=AAV2 ITR
     rep_origin      complement(4377..4965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4457..4476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5136..5927,5928..5996))
                     /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     5759..5778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5997..6101)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6127..6582)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6264..6285)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6476..6495
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc
       61 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      121 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta
      181 atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagat ctctgcagag
      241 ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct
      301 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      361 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      421 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      481 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      541 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      601 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      661 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgaattca agctgctagc
      721 aaggatccac ccgccaccat gctgcagaac gagcttgctc ttaagttggc tggacttgat
      781 attaacaaga ctggaggttc tgcccgcgag accttccagg ccttccgcac cggttctgac
      841 caactgactg aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac
      901 ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac
      961 cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca
     1021 atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa tgaaagacac agacagtgaa
     1081 gaagaaatta gagaagcgtt ccgtgtgttt gataaggatg gcaatggcta catcagtgca
     1141 gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat
     1201 gaaatgatca gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta
     1261 gtgatgatga cagcgaagga attccctcct cctccgccac cgccaccacc gccgccgccg
     1321 ccaccaccac ctcctccgcc tcctccacca ccaccgccac cgcctccacc acctggcggc
     1381 agcagggtcg actcatcacg tcgtaagttc aataagacag gtcacgcagt cagagctata
     1441 ggtcggctga gctcactcga gggcggttct gacgtcggcc gcaagctgat catcgatcag
     1501 aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag
     1561 atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc
     1621 ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac
     1681 atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt
     1741 ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg
     1801 gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag
     1861 atcgcgcgct ggctgtctac tctggagatt ggtggcaccg gtggctctgg cggtaccggt
     1921 ggctctggcg gttctatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg
     1981 ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg
     2041 cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccacatgt tgcaccgacc
     2101 catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt
     2161 tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa
     2221 gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc
     2281 aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg
     2341 gacgaatggg gaggttctat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2401 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2461 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2521 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2581 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2641 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2701 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2761 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2821 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2881 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2941 ctgctgcccg acaaccacta cctgagcacc cagtccaaac tgagcaaaga ccccaacgag
     3001 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3061 gacgagctgt acaagtgaaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     3121 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3181 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3241 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3301 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3361 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3421 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3481 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3541 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3601 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3661 ctttgggccg cctccccgca tcgataccgt cgacctcgac ccgggcggcc gcttcgagca
     3721 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3781 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3841 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3901 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
     3961 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     4021 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4081 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4141 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4201 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4261 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4321 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4381 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4441 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4501 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4561 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4621 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4681 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4741 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4801 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4861 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4921 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4981 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5041 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5101 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5161 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5221 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5281 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5341 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5401 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5461 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5521 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5581 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5641 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5701 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5761 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5821 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5881 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5941 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6001 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6061 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6121 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     6181 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     6241 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     6301 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6361 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6421 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6481 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6541 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     6601 tgcgcaactg t
//
