LOCUS       sequence-437558-        4441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sTb(N) half of CaST for neuron expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4441)
  AUTHORS   Zhang R, Anguiano M, Aarrestad IK, Lin S, Chandra J, Vadde SS, Olson
            DE, Kim CK
  TITLE     Rapid, biochemical tagging of cellular activity history in vivo.
  JOURNAL   Nat Methods. 2024 Sep;21(9):1725-1735. doi: 
            10.1038/s41592-024-02375-7. Epub 2024 Aug 5.
  PUBMED    39103446
REFERENCE   2  (bases 1 to 4441)
  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: 
            "10.1038/s41592-024-02375-7"; journalName: "Nat Methods"; date: 
            "2024-09"; volume: "21"; issue: "9"; pages: "1725-1735"
COMMENT     Sequence Label: pAAV-Syn-CaM-V5-sTb(N)
FEATURES             Location/Qualifiers
     source          1..4441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..24)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        92..196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(197..265,266..1057)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEVDKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(415..434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1228..1816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1717..1736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1878..2007
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2058..2505
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      2520..2529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3039..3080
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3339..3356)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3345..3552
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3581..3721
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3581..3710
                     /label=AAV2 ITR
     rep_origin      3796..4251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3883..3902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4093..4114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4268..4287)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4387..4409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
       61 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      181 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      241 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      301 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      361 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      421 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      481 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      541 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      601 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      661 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      721 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      781 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggtgg ataaagttgc
      841 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      901 cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      961 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1021 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1081 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1141 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1861 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     1921 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     1981 tggccaactc catcactagg ggttcctgcg gccgcacgcg ttgtacagtg tctagactgc
     2041 agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt
     2101 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
     2161 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
     2221 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
     2281 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc
     2341 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
     2401 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
     2461 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag aagaagcttg
     2521 ccaccatgga ccaactgact gaagagcaga tcgcagaatt taaagaggct ttctccctat
     2581 ttgacaagga cggggatggg acaataacaa ccaaggagct ggggacggtg atgcggtctc
     2641 tggggcagaa ccccacagaa gcagagctgc aggacatgat caatgaagta gatgccgacg
     2701 gtgacggcac aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa atgaaataca
     2761 gggacacgga agaagaaatt agagaagcgt tcggtgtgtt tgataaggat ggcaatggct
     2821 acatcagtgc agcagagctt cgccacgtga tgacaaacct tggagagaag ttaacagatg
     2881 aagaggttga tgaaatgatc agggaagcag acatcgatgg ggatggtcag gtaaactacg
     2941 aagagtttgt acaaatgatg acagcgaaga agggctcggg ctcgacctcg ggctcgggca
     3001 gcggtggctc tggaggttct ggtggcagct ctggaggtgg caagcccatc cccaaccccc
     3061 tgctgggcct ggacagcacc gctagcaaag acaatactgt gcctctgaag ctgatcgctc
     3121 tcctggctaa tggcgagttc catagtggcg aacagctggg agaaaccctg ggcatgtcca
     3181 gggccgctat caacaagcac attcagactc tgcgcgactg gggcgtggac gtgttcaccg
     3241 tgcccggaaa gggctactct ctgcccgagc ctatcccgct gctgaacgct aaacagattc
     3301 tgtaataaga attcctcgag ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3361 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3421 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3481 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3541 gcatgctggg gaggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3601 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3661 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3721 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3781 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3841 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     4141 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     4201 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4261 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4321 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4381 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4441 g
//
