LOCUS       sequence-368190-        5591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of LifeAct-jGCaMP8m under the synapsin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  TITLE     GCaMP8/jGCaMP8 AAV plasmids
REFERENCE   2  (bases 1 to 5591)
  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"
COMMENT     Sequence Label: AAV-syn-LifeAct-jGCaMP8m-WPRE
FEATURES             Location/Qualifiers
     source          1..5591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   42..171
                     /label=AAV2 ITR
     promoter        253..700
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      732..741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             738..788
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             792..809
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1091..1112
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1218..1239)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1479..1498)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2070..2658
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2123..2143)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2541..2552)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2661..2677)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2662..2678)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2707..2828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2718..2737)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2772..2791
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2990..3119
                     /label=AAV2 ITR
     rep_origin      3191..3646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3278..3297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3488..3509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3672..3776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3777..3845,3846..4637)
                     /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(3995..4014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4808..5396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5297..5316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgcgcgct cgctcgctca
       61 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
      121 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
      181 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc tctgcagagg
      241 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
      301 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
      361 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
      421 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
      481 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
      541 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
      601 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
      661 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgaattcaa gctgctagca
      721 aggatccacc cgccaccatg ggtgtggccg acctgatcaa gaagttcgag agcatcagca
      781 aggaggagat gcatcatcac catcatcaca cgcgtcgcaa gaagaccttc aaggaggtgg
      841 ccaccgccgt gaagatcatc ggccgcctga tgggcctgaa gatcaacgtc tatatcaagg
      901 ccgacaagca gaagaacggc atcaaggcga acttccacat ccgccacaac atcgaggacg
      961 gcggcgtgca gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1021 tgctgcccga caaccactac ctgagcgtgg agtccaaact ttcgaaagac cccaacgaga
     1081 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1141 acgagctgta caagggcggt accggaggga gcatggtgag caagggcgag gagctgttca
     1201 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1261 tgtccggcga gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1321 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1381 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1441 ccgaaggcta catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1501 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1561 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacctg ccggaccaac
     1621 tgactgaaga gcagatcgca gaatacaaag aggctttctc cctatttgac aaggacgggg
     1681 atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg cagaacccca
     1741 cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacaatcg
     1801 acttccctga gttcctgaca atgatggcaa gaaaaatgaa atacagggac acggaagaag
     1861 aaattagaga agcgttcggt gtgtttgata aggatggcaa tggctacatc agtgcagcag
     1921 agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag gttgatgaaa
     1981 tgatcaggga agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa
     2041 tgatgacagc gaagtaaaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2101 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2161 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2221 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2281 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2341 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2401 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2461 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2521 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2581 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2641 tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg cttcgagcag
     2701 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2761 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2821 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     2881 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     2941 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3001 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3061 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3121 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3181 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3241 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3301 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3361 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3421 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3481 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3541 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3601 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3661 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3721 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3781 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3841 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3901 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3961 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4021 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4081 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4141 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4201 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4261 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4321 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4381 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4441 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4501 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4561 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4621 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4681 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4741 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4921 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4981 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5041 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5101 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5161 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5221 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5281 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5341 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5401 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5461 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5521 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5581 gcccaatacg c
//
