LOCUS       sequence-388360-        6647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hSyn1 driven expression of the consitutively active calcium recorder
            positive control Caprola_on 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 6647)
  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 6647)
  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_on-mEGFP_WRPE-SV40
FEATURES             Location/Qualifiers
     source          1..6647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   46..175
                     /label=AAV2 ITR
     promoter        257..704
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             841..1281
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             join(2398..2400,2401..2403,2404..3117)
                     /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(2443..2464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2704..2723)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3051..3072
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3130..3718
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3183..3203)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3601..3612)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3721..3737)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3722..3738)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3767..3888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3778..3797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3832..3851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4050..4179
                     /label=AAV2 ITR
     rep_origin      complement(4416..5004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4496..4515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5175..5966,5967..6035))
                     /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     5798..5817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6036..6140)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6166..6621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6303..6324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6515..6534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
       61 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      121 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      181 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatctctgca
      241 gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg
      301 cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
      361 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
      421 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
      481 tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc
      541 accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga
      601 taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt
      661 ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat tcaagctgct
      721 agcaaggatc cacccgccac catgctgcag aacgagcttg ctcttaagtt ggctggactt
      781 gatattaaca agactggagg ttctgcccgc gagaccttcc aggccttccg caccggttct
      841 gaccaactga ctgaagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag
      901 gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag
      961 aaccccacag aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc
     1021 acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaaga cacagacagt
     1081 gaagaagaaa ttagagaagc gttccgtgtg tttgataagg atggcaatgg ctacatcagt
     1141 gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt
     1201 gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt
     1261 gtagtgatga tgacagcgaa ggaattccct cctcctccgc caccgccacc accgccgccg
     1321 ccgccaccac cacctcctcc gcctcctcca ccaccaccgc caccgcctcc accacctggc
     1381 ggcagcaggg tcgactcatc acgtcgtaag ttcaataaga caggtcacgc actgagagct
     1441 ataggtcggc tgagctcact cgagggcggt tcttttgccc gcgagacctt ccaggccttc
     1501 cgcaccaccg acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg
     1561 ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg
     1621 ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc
     1681 ggtgagccag cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc
     1741 cctgtcccga agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc
     1801 gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat
     1861 ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtctact
     1921 ctggagattg gtggcaccgg tggctctggc ggtaccggtg gctctggcgg ttctatcggt
     1981 actggctttc cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc
     2041 gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc
     2101 tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac
     2161 ctgatcggta tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc
     2221 cgcttcatgg atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac
     2281 gactggggct ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt
     2341 attgcattta tggagttcat ccgccctatc ccgacctggg acgaatgggg aggttctatg
     2401 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2461 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2521 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2581 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2641 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2701 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2761 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2821 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2881 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2941 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3001 ctgagcaccc agtccaaact gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3061 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgaaag
     3121 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3181 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3241 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3301 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3361 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3421 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3481 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3541 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3601 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3661 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3721 cgataccgtc gacctcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     3781 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3841 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3901 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     3961 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     4021 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     4081 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4141 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa
     4201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     6181 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     6241 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     6301 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     6361 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     6421 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     6481 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     6541 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     6601 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaac
//
