LOCUS       sequence-472681-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct with CKAR3 mutant expression under a human synapsin 
            promotor and WPRE3 translation enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  AUTHORS   Yahiro T, Bayless-Edwards L, Jones JA, Ma L, Qin M, Mao T, Zhong H
  TITLE     A high-performance genetically encoded sensor for cellular imaging 
            of PKC activity in vivo.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 23:2024.07.19.604387. doi: 
            10.1101/2024.07.19.604387.
  PUBMED    39091834
REFERENCE   2  (bases 1 to 5870)
  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: "bioRxiv 
            [Preprint]. 2024 Jul 23:2024.07.19.604387. doi: 
            10.1101/2024.07.19.604387."
COMMENT     Sequence Label: pAAV_mutCKAR3
FEATURES             Location/Qualifiers
     source          1..5870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   37..166
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        211..658
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      751..760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(757..759,760..762,763..1275)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     CDS             join(2191..2193,2194..2196,2197..2709)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSVNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(2789..2809)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3169..3309
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3169..3298
                     /label=AAV2 ITR
     rep_origin      3384..3839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3471..3490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3681..3702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3856..3875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3975..3997
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4035..4053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4121..4225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4226..4294,4295..5086)
                     /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(4444..4463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5257..5845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5746..5765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
       61 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
      121 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
      181 gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
      241 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      301 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      361 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      421 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      481 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      541 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      601 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      661 gagaaggtac cggatcaatt cgccaccatg ctgcagaacg agcttgctct taagttggct
      721 ggacttgata ttaacaagac tggaccggtc gccaccatgg tgagcaaggg cgaggagctg
      781 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      841 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      901 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
      961 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1021 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1081 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1141 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1201 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1261 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1321 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     1381 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1441 cgcatgcata agttttctca agaacagatc ggcgaaaaca ttgtgtgcag ggtcatttgt
     1501 accacgggtc aaattcccat ccgagatttg tcagctgata tttcacaagt gcttaaggaa
     1561 aaacgatcca taaagaaagt ttggacattt ggtagaaacc cagcctgtga ctatcattta
     1621 ggaaacattt caagactgtc aaataagcat ttccaaatac tactaggaga agacggtaac
     1681 cttttattga atgacatttc cactaatggg acctggttaa atgggcaaaa agtcgagaag
     1741 aacagcaatc agttactgtc tcaaggtgat gaaataaccg ttggtgtagg cgtggaatca
     1801 gatattttat ctctggtcat tttcataaac gacaaattta agcagtgcct cgagcagaac
     1861 aaagttgatc gctctgcagg taagccaggc agcggcgagg gcagcaccaa gggccgattc
     1921 agaagattcc aggcgttgaa ggataaggct aaggccaccg gcggcagcga gctcatgggc
     1981 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2041 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     2101 cgcgatcaca tggtcctgct ggagagagtg accgccgccg ggatcactct cggcatggac
     2161 gagctataca agggtggaac tggaggttca atggtgagca agggcgagga gctgttcacc
     2221 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2281 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct catctgcacc
     2341 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgatg
     2401 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2461 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2521 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2581 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactggaa cagcgtcaac
     2641 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2701 aacatcgagt aagcggccgc aattcccctc gagataatca acctctggat tacaaaattt
     2761 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2821 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2881 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2941 ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt gaaatttgtg
     3001 atgctattgc tttatttgta accatctagc tttatttgtg aaatttgtga tgctattgct
     3061 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3121 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcggccgcag gaacccctag
     3181 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3241 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3301 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3361 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3421 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     3481 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3541 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3601 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3661 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3721 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3781 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3841 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3901 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3961 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4021 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4081 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4141 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4201 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4261 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4321 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4381 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4441 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4501 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4561 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4621 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4681 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4741 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4801 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4861 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4921 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4981 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5041 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5101 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5161 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5221 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5281 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5341 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5401 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5461 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5521 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5581 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5641 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5701 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5761 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5821 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
//
