LOCUS       sequence-442207-        7103 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-mCHerry with soma targeting KV2.1 motif..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7103)
  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 7103)
  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 (mCherry, KV2.1)
FEATURES             Location/Qualifiers
     source          1..7103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    195..216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(231..236,237..254,255..261)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        458..905
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2090..2143
                     /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             3263..3970
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3408..3426)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3676..3695
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4230..4250)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     rep_origin      4838..5293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4925..4944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5135..5156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5319..5423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5424..5492,5493..6284)
                     /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(5642..5661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6455..7043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6944..6963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgt
      181 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      241 atgcttccgg ctcgtatgtt gtgtggaatt gtggcgcggg caaagcccgg gcgtcgggcg
      301 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      361 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      421 tgctctagga agatctctgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      481 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      541 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      601 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      661 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      721 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      781 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      841 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      901 cgcagtcgaa ttcaagcttg gatccatggc cgccctccat caggctctgg aggagctggg
      961 acagggagcc agcaggcacc tcctgtacag agcccccgag gagatggtgt atggcaagtg
     1021 ggtggaggcc aaccctaggt gcatcgaact gcacgaccac tccgactacg tgtacgtgtt
     1081 ccagctgtgc ttcgccgtgg tgtgcgcctg ccaggtgatc ttcatgttca ccagggcccc
     1141 caacgtgggc tgggaagcca tctatctgcc cctggccgag gtcgtgacct acagcattgc
     1201 cgccaacgga gagggcgtgc tgaggatggc cgacggcagg tacttcaatt tcgccaaact
     1261 ggccggatgg gccgtgtgct gccccattat gctcatccag atcggaggca tggcccagat
     1321 caagtacagg accatccccc tgaacaacgt ggtgctggcc gctagcctga ataggaccat
     1381 cttcggcatg gccagcgcca tcacagcctc cgaccctgct agatggggct tttacttctg
     1441 cgcctggatc tgttacctga ccgaggtcgg catcaccctg accatcatgg ccgtggccat
     1501 cagcgacttc agcaagatca agacagagct cggccagtgg gtggtgggcc gtattcaaac
     1561 catgaggatc atctttctgg tcgcctggac cagcttcccc gtcgtgtggg tgctgggcta
     1621 caccggattc tgcgtgatcc acgaggacta catcgccctg ctgtatctgt tcgccgacct
     1681 gctctccaag aacacctggg gagtgatgat gtggcatacc acctgggtga aactcaacgg
     1741 caaatgggac agggaatttg ccgccgccgg aggacacgag gctctgaaga aggccctgga
     1801 gcaggacgtg gaaatcggcg ccggcgagaa gaaccagaac cagctgaccg ccaggcaggc
     1861 ggccgcgcag agccagccca tcttgaatac caaggagatg gccccccaga gcaagccccc
     1921 cgaggagctg gagatgagca gcatgcccag ccccgtggcc cccctgcccg ccagaaccga
     1981 gggcgtgata gatatgaggt ctatgagcag catcgacagc ttcatctcat gtgcaactga
     2041 cttccccgag gccaccagat tcgctagcgg aggtggatct ggtggcggtg agggcagagg
     2101 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctcgtacga tggtgagcag
     2161 aagaccctgg ctgctggccc tggccctggc cgtggccctg gccgccggca gcgccggcgc
     2221 cagcaccggc agcgacgcca ccgtgcccgt ggccacccag gacggccccg actacgtgtt
     2281 ccacagagcc cacgagagaa tgctgttcca gaccagctac accctggaga acaacggcag
     2341 cgtgatctgc atccccaaca acggccagtg cttctgcctg gcctggctgc tgcacagcag
     2401 aggcacacca ggagaaaaga tcggcgccca ggtctgccag tggattgctt tcagcatcgc
     2461 catcgtcctg ctgacattct acggcttcag cgcctggaag gccacttgcg gttgggagga
     2521 ggtctacgtc tgttgcgtcg aggtgctgtt cgtgaccctg gagatcttca aggagttcag
     2581 cagccccgcc acagtgtacc tgtctaccgg caaccacgcc tattgcctgc gctacttcga
     2641 gtggctgctg tcttgccccg tgatcctgat ccgcctgagc aacctgagcg gcctgaagaa
     2701 cgactacagc aagcggacca tgggcctgat cgtgtcttgc gtgggaatga tcgtgttcgg
     2761 catggccgca ggactggcta ccgattggct caagtggctg ctgtatatcg tgtcttgcat
     2821 ctacggcggc tacatgtact tccaggccgc caagtgctac gtggaagcca accacagcgt
     2881 gcctaaaggc cattgccgca tggtcgtgaa gctgatggcc tacgctttct tcgcctcttg
     2941 gggcatgtac ccaatcctct gggcagtggg accagaagga ctgctgaagc tgagccctta
     3001 cgccaacagc atcggccaca gcatctgcga catcatcgcc aaggagtttt ggaccttcct
     3061 ggcccaccac ctgaggatca agatccacga gcacatcctg atccacggcg acatccggaa
     3121 gaccaccaag atggagatcg gaggcgagga ggtggaagtg gaagagttcg tggaggagga
     3181 ggacgaggac acagtgctcg agaagagcag gatcaccagc gagggcgagt acatccccct
     3241 ggaccagatc gacatcaacg tgatggtgag caagggcgag gaggataaca tggccatcat
     3301 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     3361 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     3421 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     3481 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     3541 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     3601 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     3661 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     3721 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     3781 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     3841 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     3901 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     3961 gctgtacaag cagtcccaac ccatactgaa cacgaaggaa atggctcctc aaagtaaacc
     4021 gcccgaagaa ctcgaaatga gttcaatgcc atcacctgtg gctccgcttc ccgcccgcac
     4081 ggaaggtgtc atcgacatgc gaagcatgtc tagtattgac tcatttatta gctgcgccac
     4141 cgatttccct gaagctactc ggttctaatc tagaataatc aacctctgga ttacaaaatt
     4201 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4261 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4321 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4381 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt
     4441 gatgctattg ctttatttgt aaccatcact agttaggctt tatttgtgaa atttgtgatg
     4501 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4561 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaaat cgataaggat
     4621 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
     4681 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4741 gcgtcagtga gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc
     4801 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4861 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4921 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4981 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5041 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5101 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5161 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5221 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5281 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5641 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6601 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7081 tttgctggcc ttttgctcac atg
//
