LOCUS       sequence-388462-        6581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hSyn1 driven expression of the inactive calcium recorder negative 
            control Caprola_off 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 6581)
  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 6581)
  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_off-mEGFP_WRPE-SV40
FEATURES             Location/Qualifiers
     source          1..6581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   54..183
                     /label=AAV2 ITR
     promoter        265..712
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             819..1259
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             1380..1436
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHALRAIGRLSS"
     CDS             join(2340..2342,2343..2345,2346..3059)
                     /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(2385..2406)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2646..2665)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2993..3014
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3072..3660
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3125..3145)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3543..3554)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3663..3679)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3664..3680)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3709..3830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3720..3739)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3774..3793
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3992..4121
                     /label=AAV2 ITR
     rep_origin      complement(4358..4946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4438..4457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5117..5908,5909..5977))
                     /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     5740..5759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5978..6082)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6108..6563)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6245..6266)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6457..6476
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
       61 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
      121 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      181 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
      241 tctctgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg
      301 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
      361 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
      421 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
      481 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
      541 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
      601 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
      661 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtcgaattc
      721 aagctgctag caaggatcca cccgccacca tgctgcagaa cgagcttgct cttaagttgg
      781 ctggacttga tattaacaag actggaggtt ctctgccgga ccaactgact gaagagcaga
      841 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa
      901 ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc
      961 aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc
     1021 tgacaatgat ggcaagaaaa atgaaagaca cagacagtga agaagaaatt agagaagcgt
     1081 tccgtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga
     1141 tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag
     1201 acatcgatgg ggatggtcag gtaaactacg aagagtttgt agtgatgatg acagcgaagg
     1261 aattccctcc tcctccgcca ccgccaccac cgccgccgcc gccaccacca cctcctccgc
     1321 ctcctccacc accaccgcca ccgcctccac cacctggcgg cagcagggtc gactcatcac
     1381 gtcgtaagtg gaataagaca ggtcacgcac tgagagctat aggtcggctg agctcactcg
     1441 agggcggttc tgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta
     1501 cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc
     1561 cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg
     1621 ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt
     1681 cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag
     1741 ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga
     1801 atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcta
     1861 ctctggagat tggtggcacc ggtggctctg gcggtaccgg tggctctggc ggttctatcg
     1921 gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg
     1981 tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct
     2041 cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag
     2101 acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg
     2161 tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc
     2221 acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag
     2281 gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg ggaggttcta
     2341 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2401 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2461 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2521 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2581 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2641 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2701 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2761 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2821 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2881 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2941 acctgagcac ccagtccaaa ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3001 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgaa
     3061 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3121 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3181 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3241 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3301 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3361 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3421 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3481 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3541 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3601 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3661 atcgataccg tcgacctcga cccgggcggc cgcttcgagc agacatgata agatacattg
     3721 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3781 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3841 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     3901 aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat
     3961 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4021 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4081 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc
     4141 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4201 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4261 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4321 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4381 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4441 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4501 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4561 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4621 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4681 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4741 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4801 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4861 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4921 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4981 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5041 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5101 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5161 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5221 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5281 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5341 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5401 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5461 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5521 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5581 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5641 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5701 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5761 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5821 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5881 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5941 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6001 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6061 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     6121 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     6181 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     6241 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     6301 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     6361 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     6421 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     6481 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     6541 taatgcgccg ctacagggcg cgtcccattc gccattcagg c
//
