LOCUS       sequence-325515-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DEPRECATED: Please see Addgene item #169249 (coming soon) for an 
            improved version of this plasmid. AAV transfer plasmid for neuronal 
            expression of soma-targeted (RL-10, ribosomal tag) jGCaMP8f..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6533)
  AUTHORS   Grodem S, Nymoen I, Vatne GH, Rogge FS, Bjornsdottir V, Lensjo KK, 
            Fyhn M
  TITLE     An updated suite of viral vectors for in vivo calcium imaging using 
            intracerebral and retro-orbital injections in male mice.
  JOURNAL   Nat Commun. 2023 Feb 4;14(1):608. doi: 10.1038/s41467-023-36324-3.
  PUBMED    36739289
REFERENCE   2  (bases 1 to 6533)
  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: "Nat 
            Commun."; date: "2023-02-4"; volume: "14(1):608. doi"; pages: " 
            10.1038/s41467-023-36324-3"
COMMENT     Sequence Label: AAV-hSyn-Ribo-jGCaMP8f
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             485..502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     784..805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(911..932)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1172..1191)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2478..3066
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2531..2551)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2949..2960)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3098..3574
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3297..3316)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3614..3754
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3614..3743
                     /label=AAV2 ITR
     rep_origin      3829..4284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3916..3935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4126..4147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4301..4320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4420..4442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4480..4498)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4566..4670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4671..4739,4740..5531)
                     /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(4889..4908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5702..6290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6191..6210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6352..6481
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg ctcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaaccgg ctagaggatc cacccgccac
      481 catgcatcat caccatcatc acacgcgtcg caagaagacc ttcaaggagg tggccaccgc
      541 cgtgaagatc atcgcccgcc tgatgggcct gaagatcaac gtctatatca aggccgacaa
      601 gcagaagaac ggcatcaagg cgaacttcca catccgccac aacatcgagg acggcggcgt
      661 gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      721 cgacaaccac tacctgagcg tggagtccaa actttcgaaa gaccccaacg agaagcgcga
      781 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
      841 gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt
      901 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      961 cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1021 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1081 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1141 ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1201 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1261 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga
     1321 agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac
     1381 aataacaacc aaggagctgg ggacggtgat gcggtctctc ggactgaacc ccacagaagc
     1441 agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc
     1501 tgagttcctg acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag
     1561 agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg
     1621 ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag
     1681 ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac
     1741 agcgaagtca ggccggactc agatctcaag ctcaagcttc gagttcagca gcaaagtctc
     1801 acgcgacacc ctgtacgagg cggtgcggga agtcctgcac gggaaccagc gcaagcgccg
     1861 caagtttctg gagacggtgg agctgcagat cagcctgaag aactacgacc ctcagaagga
     1921 caaacgtttc tcgggcaccg tcaggctcaa gtccacccca cgccccaagt tctcggtgtg
     1981 cgttctgggg gaccagcagc actgtgatga agccaaggcc gtggatatcc cccacatgga
     2041 catcgaggcg ctcaagaaac ttaacaaaaa caagaagttg gtcaagaagc tggctaagaa
     2101 gtacgatgcc tttttggcct ctgagtctct gattaagcag atcccacgta tcctgggccc
     2161 aggcctaaac aaggctggca agttcccctc cctgctgaca cacaatgaaa acatggtggc
     2221 caaagtggat gaggtgaaat cgacaatcaa gttccagatg aagaaggtgc tgtgtttggc
     2281 cgtcgctgtt ggccacgtga agatgaccga tgatgagcta gtctacaaca ttcatctggc
     2341 tgtcaatttc ttggtgtcct tgcttaagaa aaactggcaa aacgtgcggg ctctgtacat
     2401 caagagcacc atgggcaagc cccagcgtct gtattagggc gcgcccctgc agggaattcg
     2461 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2521 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2581 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2641 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2701 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2761 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2821 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     2881 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     2941 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3001 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3061 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3121 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3181 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3241 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3301 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3361 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3421 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3481 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     3541 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     3601 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3661 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3721 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3781 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3841 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3901 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3961 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4021 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4081 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4141 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4201 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4261 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4321 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4381 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4441 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4501 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     6361 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
     6421 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     6481 tgcggccgca cgcgtttaat taagtgtcta gactgcagag ggccctgcgt atg
//
