LOCUS       sequence-388359-        6611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hSyn1 driven expression of the calcium recorder Caprola_06 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_06-mEGFP_WRPE-SV40
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..134
                     /label=AAV2 ITR
     promoter        216..663
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             800..1240
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             join(2321..2323,2324..2326,2327..3040)
                     /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(2366..2387)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2627..2646)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2974..2995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3053..3641
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3106..3126)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3524..3535)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3644..3660)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3645..3661)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3690..3811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3701..3720)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3755..3774
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3973..4102
                     /label=AAV2 ITR
     rep_origin      complement(4339..4927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4419..4438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5098..5889,5890..5958))
                     /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     5721..5740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5959..6063)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6089..6544)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6226..6247)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6438..6457
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
       61 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      121 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
      181 ctctaggaag atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg
      241 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
      301 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
      361 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
      421 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
      481 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
      541 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
      601 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
      661 cagtcgaatt caagctgcta gcaaggatcc acccgccacc atgctgcaga acgagcttgc
      721 tcttaagttg gctggacttg atattaacaa gactggaggt tctgcccgcg agaccttcca
      781 ggccttccgc accggttctg accaactgac tgaagagcag atcgcagaat ttaaagaggc
      841 tttctcccta tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt
      901 gatgcggtct ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt
      961 agatgccgac ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa
     1021 aatgaaagac acagacagtg aagaagaaat tagagaagcg ttccgtgtgt ttgataagga
     1081 tggcaatggc tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa
     1141 gttaacagat gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca
     1201 ggtaaactac gaagagtttg tagtgatgat gacagcgaag gaattccctc ctcctccgcc
     1261 accgccacca ccgccgccgc cgccaccacc acctcctccg cctcctccac caccaccgcc
     1321 accgcctcca ccacctggcg gcagcagggt cgactcatca cgtcgtaagt tcaataagac
     1381 aggtcacgca ctgagagcta taggtcggct gagctcactc gagggcggtt ctgacgtcgg
     1441 ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt
     1501 ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga
     1561 ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat
     1621 cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct
     1681 gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag
     1741 cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa
     1801 cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga ttggtggcac
     1861 cggtggctct ggcggtaccg gtggctctgg cggttctatc ggtactggct ttccattcga
     1921 cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga
     1981 tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat
     2041 catcccacat gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa
     2101 atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt
     2161 catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct
     2221 gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt
     2281 catccgccct atcccgacct gggacgaatg gggaggttct atggtgagca agggcgagga
     2341 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2401 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2461 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2521 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2581 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2641 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2701 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2761 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2821 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2881 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     2941 actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3001 cgccgggatc actctcggca tggacgagct gtacaagtga aagcttatcg ataatcaacc
     3061 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3121 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3181 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3241 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3301 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3361 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3421 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3481 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3541 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3601 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     3661 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3721 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3781 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3841 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3901 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     3961 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4021 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4081 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4141 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5221 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6061 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     6121 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6181 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6241 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     6301 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6361 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     6421 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     6481 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     6541 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6601 tcgctattac g
//
