LOCUS       sequence-255902-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bicistronic AAV vector to express GCaMP6m and soma-targeted ChRmine 
            under the control of human Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  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 7080)
  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-GCaMP 6m-p2A-ChRmine-Kv2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..7080
                     /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      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             662..718
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     931..952
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1058..1079)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1319..1338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1910..1966
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3158..3184
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3212..3800
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3265..3285)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3683..3694)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3832..4308
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4031..4050)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4348..4488
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4348..4477
                     /label=AAV2 ITR
     rep_origin      4563..5018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4650..4669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4860..4881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5035..5054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5154..5176
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5214..5232)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5300..5404
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5405..5473,5474..6265)
                     /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(5623..5642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6436..7024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6925..6944
                     /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 gccaccatgg tggactcatc
      661 acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact
      721 cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttcaagat
      781 ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc
      841 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact
      901 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      961 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag
     1021 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1081 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct
     1141 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1201 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1261 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga
     1321 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1381 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1441 gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag aggctttctc
     1501 cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg
     1561 gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc
     1621 cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaagggag
     1681 ctacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata aggatggcaa
     1741 tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac
     1801 agatgaagag gttgatgaaa tgatcaggga agcagacatc gatggggatg gtcaggtaaa
     1861 ctacgaagag tttgtacaaa tgatgacagc gaagactagt ggctccggag ccacgaactt
     1921 ctctctgtta aagcaagcag gagacgtgga agaaaacccc ggtcccatgg cacacgcacc
     1981 aggcaccgac cagatgttct acgtgggcac aatggacggc tggtatctgg ataccaagct
     2041 gaactccgtg gccatcggcg cccactggtc ttgctttatc gtgctgacaa tcaccacatt
     2101 ctacctgggc tatgagtcct ggacatctcg gggccctagc aagagaacct ccttttacgc
     2161 cggctatcag gaggagcaga acctggccct gtttgtgaat ttctttgcca tgctgagcta
     2221 cttcggcaag atcgtggccg acacactggg ccacaacttc ggcgatgtgg gccccttcat
     2281 catcggcttc ggcaattaca ggtatgccga ctacatgctg acctgcccaa tgctggtgta
     2341 cgatctgctg tatcagctga gggcccccta tcgcgtgtct tgtagcgcca tcatctttgc
     2401 catcctgatg tctggcgtgc tggccgagtt ctacgcagag ggcgacccta ggctgaggaa
     2461 tggcgcctac gcctggtatg gctttggctg tttctggttt atcttcgcct actctatcgt
     2521 gatgagcatc gtggccaagc agtatagccg gctggcccag ctggcccagg atacaggcgc
     2581 cgagcacagc ctgcacgtgc tgaagttcgc cgtgtttacc ttctccatgc tgtggattct
     2641 gtttcccctg gtgtgggcca tctgccctag aggcttcggc tggatcgacg ataactggac
     2701 agaggtggcc cactgcgtgt gcgacatcgt ggccaagtcc tgttacggct ttgccctggc
     2761 ccggttcaga aagacctatg atgaggagct gtttcggctg ctggagcagc tgggacacga
     2821 cgaggatgag ttccagaagc tggagctgga tatgaggctg agcagcaatg gcgagcgcct
     2881 gcggagactg tctgcggccg ccaagagcag gatcaccagc gagggcgagt acatccccct
     2941 ggaccagatc gacatcaacg tgcagtccca gcccatcctc aacaccaagg agatggcccc
     3001 gcagagcaag cctccagagg agctggagat gagcagcatg cccagccccg tggcccctct
     3061 gcccgcacgc acggagggcg tcatcgacat gcggagcatg tccagcattg acagcttcat
     3121 cagctgtgcc acggacttcc ctgaagccac cagattctac ccatacgatg ttccagatta
     3181 cgcttaagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     3241 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3301 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3361 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3421 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3481 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3541 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3601 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3661 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3721 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3781 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3841 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3901 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3961 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     4021 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     4081 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     4141 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     4201 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     4261 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     4321 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     4381 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4441 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     4501 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4561 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4621 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4681 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4741 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4801 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4861 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4921 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4981 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     5041 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5101 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5161 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5221 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5281 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5341 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5401 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5461 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5521 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5581 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5641 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5701 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5761 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5821 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5881 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5941 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6001 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6061 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6121 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6181 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6241 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6301 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6361 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6421 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6481 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6541 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6601 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6661 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6721 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6781 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6841 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6901 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6961 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7021 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
//
