LOCUS       sequence-368189-        5588 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV-mediated cytosolic expression of jGCaMP8m under the synapsin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5588)
  TITLE     GCaMP8/jGCaMP8 AAV plasmids
REFERENCE   2  (bases 1 to 5588)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AAV-syn-NES-jGCaMP8m-WPRE
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   70..199
                     /label=AAV2 ITR
     promoter        281..728
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      760..769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             817..834
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1116..1137
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1243..1264)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1504..1523)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2095..2683
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2148..2168)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2566..2577)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2686..2702)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2687..2703)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2732..2853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2743..2762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2797..2816
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3015..3144
                     /label=AAV2 ITR
     rep_origin      complement(3381..3969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3461..3480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4140..4931,4932..5000))
                     /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     4763..4782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5001..5105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5131..5586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5268..5289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5480..5499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
       61 ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
      121 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
      181 tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag
      241 ccatgctcta ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg ggttttagga
      301 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
      361 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      421 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      481 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      541 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      601 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      661 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      721 gagcgcagtc gaattcaagc tgctagcaag gatccacccg ccaccatggg taacctgaac
      781 gaactggcat tgaagttggc cggtttggac atcatgcatc atcaccatca tcacacgcgt
      841 cgcaagaaga ccttcaagga ggtggccacc gccgtgaaga tcatcggccg cctgatgggc
      901 ctgaagatca acgtctatat caaggccgac aagcagaaga acggcatcaa ggcgaacttc
      961 cacatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta ccagcagaac
     1021 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cgtggagtcc
     1081 aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1141 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg
     1201 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1261 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc
     1321 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1381 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1441 cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac catcttcttc
     1501 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1561 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1621 ctggagtaca acctgccgga ccaactgact gaagagcaga tcgcagaata caaagaggct
     1681 ttctccctat ttgacaagga cggggatggg acaataacaa ccaaggagct ggggacggtg
     1741 atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat caatgaagta
     1801 gatgccgacg gtgacggcac aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa
     1861 atgaaataca gggacacgga agaagaaatt agagaagcgt tcggtgtgtt tgataaggat
     1921 ggcaatggct acatcagtgc agcagagctt cgccacgtga tgacaaacct tggagagaag
     1981 ttaacagatg aagaggttga tgaaatgatc agggaagcag acatcgatgg ggatggtcag
     2041 gtaaactacg aagagtttgt acaaatgatg acagcgaagt aaaagcttat cgataatcaa
     2101 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2161 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2221 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2281 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2341 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2401 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2461 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     2521 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2581 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2641 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     2701 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2761 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2821 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2881 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2941 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     3001 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3061 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3121 ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3181 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3781 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5101 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     5161 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5221 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5281 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5341 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5401 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5461 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5521 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5581 gcgcgtcc
//
