LOCUS       sequence-441799-        7082 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A recombinant AAV2 plasmid encoding the ZipACR I151V 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 7082)
  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 7082)
  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-ZipV-2A-IvfChr (citrine, KV2.1)
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(216..804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(296..315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(975..1766,1767..1835))
                     /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     1598..1617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1836..1940)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1966..2421)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2103..2124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2315..2334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2493..2622
                     /label=AAV2 ITR
     promoter        2704..3151
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4336..4389
                     /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(5509..5511,5512..5514,5515..6225)
                     /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(5554..5575)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5815..5834)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6162..6183
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6485..6505)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   6932..7061
                     /label=AAV2 ITR
ORIGIN
        1 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
       61 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1261 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1561 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1681 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     1981 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2041 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2101 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2161 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2221 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2281 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2341 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2401 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     2461 gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc actgaggccg
     2521 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     2581 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
     2641 cgccatgcta cttatctacg tagccatgct ctaggaagat ctctgcagag ggccctgcgt
     2701 atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac
     2761 cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc
     2821 agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc
     2881 gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg
     2941 cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt
     3001 ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg
     3061 gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag
     3121 cggaggagtc gtgtcgtgcc tgagagcgca gtcgaattca agcttggatc catggccgcc
     3181 ctccatcagg ctctggagga gctgggacag ggagccagca ggcacctcct gtacagagcc
     3241 cccgaggaga tggtgtatgg caagtgggtg gaggccaacc ctaggtgcat cgaactgcac
     3301 gaccactccg actacgtgta cgtgttccag ctgtgcttcg ccgtggtgtg cgcctgccag
     3361 gtgatcttca tgttcaccag ggcccccaac gtgggctggg aagccatcta tctgcccctg
     3421 gccgaggtcg tgacctacag cattgccgcc aacggagagg gcgtgctgag gatggccgac
     3481 ggcaggtact tcaatttcgc caaactggcc ggatgggccg tgtgctgccc cattatgctc
     3541 atccagatcg gaggcatggc ccagatcaag tacaggacca tccccctgaa caacgtggtg
     3601 ctggccgcta gcctgaatag ggtcatcttc ggcatggcca gcgccatcac agcctccgac
     3661 cctgctagat ggggctttta cttctgcgcc tggatctgtt acctgaccga ggtcggcatc
     3721 accctgacca tcatggccgt ggccatcagc gacttcagca agatcaagac agagctcggc
     3781 cagtgggtgg tgggccgtat tcaaaccatg aggatcatct ttctggtcgc ctggaccagc
     3841 ttccccgtcg tgtgggtgct gggctacacc ggattctgcg tgatccacga ggactacatc
     3901 gccctgctgt atctgttcgc cgacctgctc tccaagaaca cctggggagt gatgatgtgg
     3961 cataccacct gggtgaaact caacggcaaa tgggacaggg aatttgccgc cgccggagga
     4021 cacgaggctc tgaagaaggc cctggagcag gacgtggaaa tcggcgccgg cgagaagaac
     4081 cagaaccagc tgaccgccag gcaggcggcc gcgcagagcc agcccatctt gaataccaag
     4141 gagatggccc cccagagcaa gccccccgag gagctggaga tgagcagcat gcccagcccc
     4201 gtggcccccc tgcccgccag aaccgagggc gtgatagata tgaggtctat gagcagcatc
     4261 gacagcttca tctcatgtgc aactgacttc cccgaggcca ccagattcgc tagcggaggt
     4321 ggatctggtg gcggtgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     4381 cccggccctc gtacgatggt gagcagaaga ccctggctgc tggccctggc cctggccgtg
     4441 gccctggccg ccggcagcgc cggcgccagc accggcagcg acgccaccgt gcccgtggcc
     4501 acccaggacg gccccgacta cgtgttccac agagcccacg agagaatgct gttccagacc
     4561 agctacaccc tggagaacaa cggcagcgtg atctgcatcc ccaacaacgg ccagtgcttc
     4621 tgcctggcct ggctgctgca cagcagaggc acaccaggag aaaagatcgg cgcccaggtc
     4681 tgccagtgga ttgctttcag catcgccatc gtcctgctga cattctacgg cttcagcgcc
     4741 tggaaggcca cttgcggttg ggaggaggtc tacgtctgtt gcgtcgaggt gctgttcgtg
     4801 accctggaga tcttcaagga gttcagcagc cccgccacag tgtacctgtc taccggcaac
     4861 cacgcctatt gcctgcgcta cttcgagtgg ctgctgtctt gccccgtgat cctgatccgc
     4921 ctgagcaacc tgagcggcct gaagaacgac tacagcaagc ggaccatggg cctgatcgtg
     4981 tcttgcgtgg gaatgatcgt gttcggcatg gccgcaggac tggctaccga ttggctcaag
     5041 tggctgctgt atatcgtgtc ttgcatctac ggcggctaca tgtacttcca ggccgccaag
     5101 tgctacgtgg aagccaacca cagcgtgcct aaaggccatt gccgcatggt cgtgaagctg
     5161 atggcctacg ctttcttcgc ctcttggggc atgtacccaa tcctctgggc agtgggacca
     5221 gaaggactgc tgaagctgag cccttacgcc aacagcatcg gccacagcat ctgcgacatc
     5281 atcgccaagg agttttggac cttcctggcc caccacctga ggatcaagat ccacgagcac
     5341 atcctgatcc acggcgacat ccggaagacc accaagatgg agatcggagg cgaggaggtg
     5401 gaagtggaag agttcgtgga ggaggaggac gaggacacag tgctcgagaa gagcaggatc
     5461 accagcgagg gcgagtacat ccccctggac cagatcgaca tcaacgtgat ggtgagcaag
     5521 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     5581 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     5641 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     5701 ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     5761 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     5821 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     5881 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     5941 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     6001 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     6061 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     6121 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     6181 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcagtc ccaacccata
     6241 ctgaacacga aggaaatggc tcctcaaagt aaaccgcccg aagaactcga aatgagttca
     6301 atgccatcac ctgtggctcc gcttcccgcc cgcacggaag gtgtcatcga catgcgaagc
     6361 atgtctagta ttgactcatt tattagctgc gccaccgatt tccctgaagc tactcggttc
     6421 taatctagaa taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6481 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6541 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     6601 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6661 ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     6721 tcactagtta ggctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     6781 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     6841 ggagatgtgg gaggtttttt aaaatcgata aggatcttcc tagagcatgg ctacgtagat
     6901 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     6961 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     7021 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc
     7081 aa
//
