LOCUS       sequence-354036-        5980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of GCaMP8s under the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5980)
  TITLE     jGCaMP8 expression under the CAG promoter
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5980)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAV-CAG-jGCaMP8s-WPRE
FEATURES             Location/Qualifiers
     source          1..5980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        485..762
                     /label=chicken beta-actin promoter
     primer_bind     1041..1063
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1125..1144
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1190..1207
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1489..1510
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1616..1637)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1877..1896)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2468..3056
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2521..2541)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2939..2950)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3099..3220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3110..3129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3164..3183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3382..3511
                     /label=AAV2 ITR
     rep_origin      complement(3748..4336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3828..3847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4507..5298,5299..5367))
                     /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     5130..5149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5368..5472)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5498..5953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5635..5656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5847..5866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag agtaccattg
      241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      481 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      541 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
      601 ggggggcgcg cgccrggsgg ggsggggsgg ggsgrggggs ggggsggggc gaggcggaga
      661 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      721 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      781 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      841 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      901 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      961 gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg
     1021 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1081 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1141 tgtgctgtct catcattttg gcaaagaatt ggatccaccc gccaccatgc atcatcacca
     1201 tcatcacacg cgtcgcaaga agaccttcaa ggaggtggcc accgccgtga agatcatcgc
     1261 catgctgatg ggcctgaaga tcaacgtcta tatcaaggcc gacaagcaga agaacggcat
     1321 caaggcgaac ttccacatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca
     1381 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1441 gagcgtggag tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1501 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac
     1561 cggagggagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1621 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga
     1681 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1741 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1801 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg
     1861 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1921 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1981 cctggggcac aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga
     2041 atacaaagag gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga
     2101 gctggggacg gtgatgcggt ctctcggaca caaccccaca gaagcagagc tgcaggacat
     2161 gatcaatgaa gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat
     2221 gatggcaaga aaaatgaaat acagggacac ggaagaagaa attagagaag cgttcggtgt
     2281 gtttgataag gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa
     2341 ccttggagag aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga
     2401 tggggatggt caggtaaact acgaagagtt tgtacaaatg atgacagcga agtaaaagct
     2461 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2521 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2581 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2641 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2701 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2761 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2821 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2881 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2941 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3001 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3061 ataccgtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3121 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3181 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3241 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     3301 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     3361 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     3421 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3481 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg
     3541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5461 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     5521 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5581 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5641 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5701 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5761 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5821 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5881 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5941 ctacagggcg cgtcccattc gccattcagg ctgcgcaact
//
