LOCUS       sequence-434252-        6130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sTb(C) half of CaST for neuron expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6130)
  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 6130)
  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-CD4-sTb(C)-M13-GFP
FEATURES             Location/Qualifiers
     source          1..6130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(86..103)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    92..299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   328..468
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   328..457
                     /label=AAV2 ITR
     rep_origin      543..998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(630..649)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     840..861
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1015..1034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1134..1156
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1194..1212)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1280..1384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1385..1453,1454..2245)
                     /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"
     rep_origin      2416..3004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2905..2924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3066..3195
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3246..3693
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3708..3717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3714..3773
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             3798..3824
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3825..3851
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5376..5432
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             join(5463..5465,5466..5468,5469..6130,1..52)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5463..5465,5466..5468,5469..6130,1..49)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5769..5788)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     join(6116..6130,1..7)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aataagaatt
       61 cctcgagcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
      121 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
      181 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
      241 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggag
      301 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
      361 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
      421 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
      481 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
      541 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      601 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      661 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      721 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
      781 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      841 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
      901 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
      961 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     1021 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     1081 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     1141 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     1201 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1261 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1321 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1381 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1441 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1501 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1561 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1621 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1681 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1741 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1801 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1861 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1921 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1981 agcttcccgg caacaattaa tagactggat ggaggtggat aaagttgcag gaccacttct
     2041 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2101 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2161 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2221 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2281 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2341 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2401 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2461 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2521 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2581 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2641 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2701 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2761 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2821 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2881 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2941 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3001 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3061 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
     3121 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     3181 tcactagggg ttcctgcggc cgcacgcgtt gtacagtgtc tagactgcag agggccctgc
     3241 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
     3301 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
     3361 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
     3421 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
     3481 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
     3541 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
     3601 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
     3661 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa gaagcttgcc accatggaga
     3721 cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacgggg
     3781 cccagccggc cagatcttac ccatacgatg tgccagatta cgcctatccc tatgacgtcc
     3841 ctgattacgc tctcgacttc cagaaggcct ccagcatagt ctataagaaa gagggggaac
     3901 aggtggagtt ctccttccca ctcgccttta cagttgaaaa gctgacgggc agtggcgagc
     3961 tgtggtggca ggcggagagg gcttcctcct ccaagtcttg gatcaccttt gacctgaaga
     4021 acaaggaagt gtctgtaaaa cgggttaccc aggaccctaa gctccagatg ggcaagaagc
     4081 tcccgctcca cctcaccctg ccccaggcct tgcctcagta tgctggctct ggaaacctca
     4141 ccctggccct tgaagcgaaa acaggaaagt tgcatcagga agtgaacctg gtggtgatga
     4201 gagccactca gctccagaaa aatttgacct gtgaggtgtg gggacccacc tcccctaagc
     4261 tgatgctgag cttgaaactg gagaacaagg aggcaaaggt ctcgaagcgg gagaaggcgg
     4321 tgtgggtgct gaaccctgag gcggggatgt ggcagtgtct gctgagtgac tcgggacagg
     4381 tcctgctgga atccaacatc aaggttctgc ccacatggtc caccccggtg cagccaatgg
     4441 ccctgattgt gctggggggc gtcgccggcc tcctgctttt cattgggcta ggcatcttct
     4501 tctgtgtcag gtgccggcac cgaaggcgca agggctcggg ctcgacctcg ggctcgggca
     4561 gcggtggctc tagaggttct ggtggcagct ctggaggtgg acagctggac ggcgggagcg
     4621 tggcagtcct gcctgtggtc gactccacca atcagtacct gctggatcga atcggcgagc
     4681 tgaagagtgg ggatgcttgc attgcagaat atcagcaggc agggagagga agcagaggga
     4741 ggaaatggtt ctctcctttt ggagctaacc tgtacctgag tatgttttgg cgcctgaagc
     4801 ggggaccagc agcaatcggc ctgggcccgg tcatcggaat tgtcatggca gaagcgctgc
     4861 gaaagctggg agcagacaag gtgcgagtca aatggcccaa tgacctgtat ctgcaggata
     4921 gaaagctggc aggcatcctg gtggagctgg ccggaataac aggcgatgct gcacagatcg
     4981 tcattggcgc cgggattaac gtggctatga ggcgcgtgga ggaaagcgtg gtcaatcagg
     5041 gctggatcac actgcaggaa gcagggatta acctggacag gaatactctg gccgctacgc
     5101 tgatccgaga gctgcgggca gccctggaac tgttcgagca ggaaggcctg gctccatatc
     5161 tgccacggtg ggagaagctg gataacttca tcaatagacc cgtgaagctg atcattgggg
     5221 acaaagagat tttcgggatt agccggggga ttgataaaca gggagccctg ctgctggaac
     5281 aggacggagt tatcaaaccc tggatgggcg gagaaatcag tctgcggtct gccgaaaagg
     5341 gctcgggcgg gagtaccggt ggtagtggga gcggtcgtcg taagtggaat aagacaggtc
     5401 acgcagtcag agctataggt cggctgagct caggtggaag tggatcaggc agcggtggat
     5461 ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     5521 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     5581 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     5641 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     5701 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     5761 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     5821 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     5881 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     5941 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     6001 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     6061 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     6121 tcctgctgga
//
