LOCUS       sequence-442289-        7083 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A recombinant AAV2 plasmid encoding the ZipACR I151T mutant with 
            soma targeting with KV2.1 motif and membrane expression enhanced 
            vfChrimson-Citrine with soma targeting KV2.1 motif..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Mermet-Joret N, Moreno A, Zbela A, Ellendersen BE, Krauth N, von 
            Philipsborn A, Piriz J, Lin JY, Nabavi S
  TITLE     Dual-color optical activation and suppression of neurons with high 
            temporal precision
  JOURNAL   eLife 2023 12:RP90327
REFERENCE   2  (bases 1 to 7083)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife 2023 
            12:RP90327"
COMMENT     Sequence Label: AAV-hSyn-ZipT-2A-IvfChr (citrine, KV2.1)
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(231..819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(311..330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(990..1781,1782..1850))
                     /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     1613..1632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1851..1955)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1981..2436)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2118..2139)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2330..2349
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2508..2637
                     /label=AAV2 ITR
     promoter        2719..3167
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4352..4405
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(5525..5527,5528..5530,5531..6241)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5570..5591)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5831..5850)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6178..6199
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6501..6521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   6948..7077
                     /label=AAV2 ITR
ORIGIN
        1 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
       61 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      841 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      901 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      961 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1021 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1081 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1141 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1201 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1261 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1321 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1381 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1441 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1501 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1561 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1621 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1681 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1741 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1801 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1861 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1981 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2041 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2101 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2161 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2221 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2281 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2341 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2401 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     2461 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct
     2521 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
     2581 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
     2641 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg aagatctctg
     2701 cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggkggggg
     2761 tgncctacct gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa
     2821 ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac
     2881 tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc
     2941 gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg
     3001 gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg
     3061 agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc
     3121 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg aattcaagct
     3181 tggatccatg gccgccctcc atcaggctct ggaggagctg ggacagggag ccagcaggca
     3241 cctcctgtac agagcccccg aggagatggt gtatggcaag tgggtggagg ccaaccctag
     3301 gtgcatcgaa ctgcacgacc actccgacta cgtgtacgtg ttccagctgt gcttcgccgt
     3361 ggtgtgcgcc tgccaggtga tcttcatgtt caccagggcc cccaacgtgg gctgggaagc
     3421 catctatctg cccctggccg aggtcgtgac ctacagcatt gccgccaacg gagagggcgt
     3481 gctgaggatg gccgacggca ggtacttcaa tttcgccaaa ctggccggat gggccgtgtg
     3541 ctgccccatt atgctcatcc agatcggagg catggcccag atcaagtaca ggaccatccc
     3601 cctgaacaac gtggtgctgg ccgctagcct gaataggacc atcttcggca tggccagcgc
     3661 catcacagcc tccgaccctg ctagatgggg cttttacttc tgcgcctgga tctgttacct
     3721 gaccgaggtc ggcatcaccc tgaccatcat ggccgtggcc atcagcgact tcagcaagat
     3781 caagacagag ctcggccagt gggtggtggg ccgtattcaa accatgagga tcatctttct
     3841 ggtcgcctgg accagcttcc ccgtcgtgtg ggtgctgggc tacaccggat tctgcgtgat
     3901 ccacgaggac tacatcgccc tgctgtatct gttcgccgac ctgctctcca agaacacctg
     3961 gggagtgatg atgtggcata ccacctgggt gaaactcaac ggcaaatggg acagggaatt
     4021 tgccgccgcc ggaggacacg aggctctgaa gaaggccctg gagcaggacg tggaaatcgg
     4081 cgccggcgag aagaaccaga accagctgac cgccaggcag gcggccgcgc agagccagcc
     4141 catcttgaat accaaggaga tggcccccca gagcaagccc cccgaggagc tggagatgag
     4201 cagcatgccc agccccgtgg cccccctgcc cgccagaacc gagggcgtga tagatatgag
     4261 gtctatgagc agcatcgaca gcttcatctc atgtgcaact gacttccccg aggccaccag
     4321 attcgctagc ggaggtggat ctggtggcgg tgagggcaga ggaagtcttc taacatgcgg
     4381 tgacgtggag gagaatcccg gccctcgtac gatggtgagc agaagaccct ggctgctggc
     4441 cctggccctg gccgtggccc tggccgccgg cagcgccggc gccagcaccg gcagcgacgc
     4501 caccgtgccc gtggccaccc aggacggccc cgactacgtg ttccacagag cccacgagag
     4561 aatgctgttc cagaccagct acaccctgga gaacaacggc agcgtgatct gcatccccaa
     4621 caacggccag tgcttctgcc tggcctggct gctgcacagc agaggcacac caggagaaaa
     4681 gatcggcgcc caggtctgcc agtggattgc tttcagcatc gccatcgtcc tgctgacatt
     4741 ctacggcttc agcgcctgga aggccacttg cggttgggag gaggtctacg tctgttgcgt
     4801 cgaggtgctg ttcgtgaccc tggagatctt caaggagttc agcagccccg ccacagtgta
     4861 cctgtctacc ggcaaccacg cctattgcct gcgctacttc gagtggctgc tgtcttgccc
     4921 cgtgatcctg atccgcctga gcaacctgag cggcctgaag aacgactaca gcaagcggac
     4981 catgggcctg atcgtgtctt gcgtgggaat gatcgtgttc ggcatggccg caggactggc
     5041 taccgattgg ctcaagtggc tgctgtatat cgtgtcttgc atctacggcg gctacatgta
     5101 cttccaggcc gccaagtgct acgtggaagc caaccacagc gtgcctaaag gccattgccg
     5161 catggtcgtg aagctgatgg cctacgcttt cttcgcctct tggggcatgt acccaatcct
     5221 ctgggcagtg ggaccagaag gactgctgaa gctgagccct tacgccaaca gcatcggcca
     5281 cagcatctgc gacatcatcg ccaaggagtt ttggaccttc ctggcccacc acctgaggat
     5341 caagatccac gagcacatcc tgatccacgg cgacatccgg aagaccacca agatggagat
     5401 cggaggcgag gaggtggaag tggaagagtt cgtggaggag gaggacgagg acacagtgct
     5461 cgagaagagc aggatcacca gcgagggcga gtacatcccc ctggaccaga tcgacatcaa
     5521 cgtgatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     5581 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     5641 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     5701 caccctcgtg accaccttcg gctacggcct gatgtgcttc gcccgctacc ccgaccacat
     5761 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     5821 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     5881 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     5941 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     6001 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     6061 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     6121 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     6181 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     6241 gcagtcccaa cccatactga acacgaagga aatggctcct caaagtaaac cgcccgaaga
     6301 actcgaaatg agttcaatgc catcacctgt ggctccgctt cccgcccgca cggaaggtgt
     6361 catcgacatg cgaagcatgt ctagtattga ctcatttatt agctgcgcca ccgatttccc
     6421 tgaagctact cggttctaat ctagaataat caacctctgg attacaaaat ttgtgaaaga
     6481 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6541 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6601 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     6661 gctcggctgt tgggcactga caattccgtg gtgtttattt gtgaaatttg tgatgctatt
     6721 gctttatttg taaccatcac tagttaggct ttatttgtga aatttgtgat gctattgctt
     6781 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     6841 tgtttcaggt tcagggggag atgtgggagg ttttttaaaa tcgataagga tcttcctaga
     6901 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     6961 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     7021 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     7081 cat
//
