LOCUS       sequence-477104-        5761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid for Syn-mediated expression of FRCaMPi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5761)
  AUTHORS   Zhou S, Zhu Q, Eom M, Fang S, Subach OM, Ran C, Alvarado JS, 
            Sunkavalli PS, Dong Y, Wang Y, Hu J, Zhang H, Wang Z, Sun X, Yang T,
            Mu Y, Yoon YG, Guo ZV, Subach FV, Piatkevich KD
  TITLE     A sensitive soma-localized red fluorescent calcium indicator for in 
            vivo imaging of neuronal populations at single-cell resolution.
  JOURNAL   PLoS Biol. 2025 Apr 29;23(4):e3003048. doi: 
            10.1371/journal.pbio.3003048. eCollection 2025 Apr.
  PUBMED    40299972
REFERENCE   2  (bases 1 to 5761)
  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"; journalName: "PLoS 
            Biol."; date: "2025-04-29"; volume: "23(4):e3003048. doi"; pages: " 
            10.1371/journal.pbio.3003048. eCollection 2025 Apr"
COMMENT     Sequence Label: pAAV-Syn-NES-FRCaMPi
FEATURES             Location/Qualifiers
     source          1..5761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        349..796
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             827..853
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     1276..1295
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2159..2747
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2212..2232)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2630..2641)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2765..2972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3024..3164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3024..3153
                     /label=AAV2 ITR
     rep_origin      3239..3694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3326..3345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3536..3557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3711..3730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3830..3852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3890..3908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3976..4080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4081..4149,4150..4941)
                     /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     complement(4299..4318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5112..5700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5601..5620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aattaattaa tggttaatta atctagactg cagagggccc tgcgtatgag tgcaagtggg
      361 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
      421 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
      481 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
      541 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
      601 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
      661 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
      721 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
      781 gtgcctgaga gcgcagtcga gaggatccta agccaccatg cttcaacttc ctcctcttga
      841 acgtcttact ctttcgagat ctatggtgag caagggcgag gaggataaca tggccatcat
      901 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      961 gatcgagggc gagggcgagg gccgccccta cgaggccttc cagaccgcta ggctgaaggt
     1021 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     1081 ctccaaggcc tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc
     1141 cgagggcttc aggtgggagc gcgtgatgag cttcgaggac ggcggcatta ttcacgttaa
     1201 tcaggactcc tccctgcagg acggcgtatt catctacaag gtgaagctgc tcggcaccaa
     1261 cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg cttcctacga
     1321 gatgactacc cgtaacctta cagatgagca gattgcggag ttccgtgagg ccttttcgct
     1381 gcttgatcgt gatcaggatg gaaatatcac gtccaatgaa ttgggtgtgg ttatgaggtc
     1441 gttaggtcta tcgcctactg ccgccgaatt acaagatatg attaatgagg tcgatgccga
     1501 tggtaatggc acaattgatt ttaccgaatt tttgacaatg atggcccgga aaatgaagga
     1561 taccgacgac gaagaggaag ttcgcgaagc ctttaaagtc ttcgataaag atggaagtgg
     1621 atacattaca gtcgaggagc tgactcatgt tcttacaagt ctcggtgaac gtttgtctcg
     1681 agaagaagta gccgatgtga tacgtgaagc cgactccgat ggcgatggtg taatcaacta
     1741 cgaagaattt tctcgtgtca tttcttccaa gggcggttca ggaggcagcg ggtcaggttc
     1801 aatgaggaaa ccgttccgtg gcgcgggcaa cgctgtgcgt gctttcagca cttggaaaaa
     1861 gctagtgccg cgggtttccg agtggatgta ccccgaggac ggcgccctga agagcgagat
     1921 caggaagggg ctgaggctga aggacggcgg ccactacgcc gccgaggtca agaccaccta
     1981 caaggccaag aagcccgtgc agctgcccgg cgcctacatc gtcgacatcg agttggacat
     2041 cttgtcccac aacgaggact acaccatcgt ggaacagtgc gaacgcgccg tgggccgcca
     2101 ccccaccggt ggcacggacg cgctgtacaa gtaggaattc cgtacaaagc ttatcgataa
     2161 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2221 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2281 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2341 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2401 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2461 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2521 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2581 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2641 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2701 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccggct
     2761 cgagctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2821 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2881 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2941 gattgggaag acaatagcag gcatgctggg gatgcggtct cgagctgatt ttgtaggtaa
     3001 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     3061 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     3121 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     3181 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     3241 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3301 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3361 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3421 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     3481 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3541 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     3601 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3661 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3721 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3781 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3841 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3901 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3961 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4021 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4081 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4141 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4201 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4261 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4321 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4381 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4441 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4501 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4561 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4621 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4681 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4741 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4801 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4861 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4921 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4981 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5041 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5221 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5461 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5641 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5761 t
//
