LOCUS       sequence-255906-        6480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of soma-targeted ChRmine-eYFP 
            under the control of human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6480)
  AUTHORS   Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja 
            C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, 
            Yoshizawa S, Kato HE, Ganguli S, Deisseroth K
  TITLE     Cortical layer-specific critical dynamics triggering perception.
  JOURNAL   Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 
            10.1126/science.aaw5202. Epub 2019 Jul 18.
  PUBMED    31320556
REFERENCE   2  (bases 1 to 6480)
  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: "Science."; 
            date: "2019-08-9"; volume: "365(6453). pii: science.aaw5202. doi"; 
            pages: " 10.1126/science.aaw5202. Epub 2019 Jul 18"
COMMENT     Sequence Label: pAAV-hSyn-ChRmine-eYFP-Kv2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..6480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        180..627
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      653..662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1655..1657,1658..2368)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1697..1718)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1958..1977)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2305..2326
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2612..3200
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2665..2685)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3083..3094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3232..3708
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3431..3450)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3748..3888
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3748..3877
                     /label=AAV2 ITR
     rep_origin      3963..4418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4050..4069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4260..4281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4435..4454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4554..4576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4614..4632)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4700..4804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4805..4873,4874..5665)
                     /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(5023..5042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5836..6424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6325..6344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggatcc ccgggtaccg gtgccaccat
      661 ggcacacgca ccaggcaccg accagatgtt ctacgtgggc acaatggacg gctggtatct
      721 ggataccaag ctgaactccg tggccatcgg cgcccactgg tcttgcttta tcgtgctgac
      781 aatcaccaca ttctacctgg gctatgagtc ctggacatct cggggcccta gcaagagaac
      841 ctccttttac gccggctatc aggaggagca gaacctggcc ctgtttgtga atttctttgc
      901 catgctgagc tacttcggca agatcgtggc cgacacactg ggccacaact tcggcgatgt
      961 gggccccttc atcatcggct tcggcaatta caggtatgcc gactacatgc tgacctgccc
     1021 aatgctggtg tacgatctgc tgtatcagct gagggccccc tatcgcgtgt cttgtagcgc
     1081 catcatcttt gccatcctga tgtctggcgt gctggccgag ttctacgcag agggcgaccc
     1141 taggctgagg aatggcgcct acgcctggta tggctttggc tgtttctggt ttatcttcgc
     1201 ctactctatc gtgatgagca tcgtggccaa gcagtatagc cggctggccc agctggccca
     1261 ggatacaggc gccgagcaca gcctgcacgt gctgaagttc gccgtgttta ccttctccat
     1321 gctgtggatt ctgtttcccc tggtgtgggc catctgccct agaggcttcg gctggatcga
     1381 cgataactgg acagaggtgg cccactgcgt gtgcgacatc gtggccaagt cctgttacgg
     1441 ctttgccctg gcccggttca gaaagaccta tgatgaggag ctgtttcggc tgctggagca
     1501 gctgggacac gacgaggatg agttccagaa gctggagctg gatatgaggc tgagcagcaa
     1561 tggcgagcgc ctgcggagac tgtctgcggc cgccaagagc aggatcacca gcgagggcga
     1621 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac
     1681 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1741 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1801 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
     1861 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1921 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1981 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2041 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2101 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2161 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2221 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     2281 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2341 cactctcggc atggacgagc tgtacaagca gtcccagccc atcctcaaca ccaaggagat
     2401 ggccccgcag agcaagcctc cagaggagct ggagatgagc agcatgccca gccccgtggc
     2461 ccctctgccc gcacgcacgg agggcgtcat cgacatgcgg agcatgtcca gcattgacag
     2521 cttcatcagc tgtgccacgg acttccctga agccaccaga ttcttctgct acgagaacga
     2581 ggtgtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2641 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3001 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3121 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3181 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3241 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3301 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3361 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3421 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3481 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3541 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3601 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3661 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3721 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3781 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3841 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3901 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3961 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4021 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4141 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4261 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4321 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4441 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4501 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4561 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4621 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4681 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4741 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4801 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4861 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4921 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4981 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5041 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5101 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5161 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5221 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5281 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5341 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5401 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5461 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5521 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5581 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5641 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5701 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5761 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5821 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5881 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5941 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6001 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6061 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6121 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6181 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6241 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6301 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6361 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6421 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
//
