LOCUS       sequence-241952-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optopatch4. Simultaneous optical recording and manipulation of 
            electrical activity in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7537)
  AUTHORS   Fan LZ, Kheifets S, Bohm UL, Wu H, Piatkevich KD, Xie ME, Parot V, 
            Ha Y, Evans KE, Boyden ES, Takesian AE, Cohen AE
  TITLE     All-Optical Electrophysiology Reveals the Role of Lateral Inhibition
            in Sensory Processing in Cortical Layer 1.
  JOURNAL   Cell. 2020 Jan 23. pii: S0092-8674(20)30047-7. doi: 
            10.1016/j.cell.2020.01.001.
  PUBMED    31978320
REFERENCE   2  (bases 1 to 7537)
  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: "Cell. 2020 
            Jan 23. pii: S0092-8674(20)30047-7. doi: 
            10.1016/j.cell.2020.01.001."
COMMENT     Sequence Label: LZF1735 pAAV_hSyn-DiO-somArchon1-EGFP-P2A-somC...
FEATURES             Location/Qualifiers
     source          1..7537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(671..704)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(755..788)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(995..1021)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1037..1981)
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="GGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQC
                     SSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKVI
                     LEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLV
                     SDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLVL
                     AMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIH
                     EHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     CDS             complement(1982..2038)
                     /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             complement(join(2288..2998,2999..3001))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(2288..2998,2999..3001))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2330..2351)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2679..2698
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2938..2959
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3865..3874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3884..3917
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3968..4001
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4026..4614
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4079..4099)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4497..4508)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4668..4687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4722..4741
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4800..4940
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4800..4929
                     /label=AAV2 ITR
     rep_origin      5015..5470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5102..5121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5312..5333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5487..5506)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5606..5628
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5666..5684)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5752..5856
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5857..5925,5926..6717)
                     /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(6075..6094)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6888..7476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7377..7396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      721 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      781 gaagttatgg cgcgccttag aatcgagtag cctctggaaa atcggtggcg catgaaatga
      841 agctgtcgat actggacata gatcgcatgt ctataacgcc ttcagtccgt gctggaaggg
      901 gtgcgacagg gctgggcatg gatgacatct ccaattcctc gggtggtttt gattgtgggg
      961 ccatttcctt agtgttaagt ataggctgac tctgagcgta atctggaaca tcgtatgggt
     1021 agctcttggc ggccgccact ttatcttcct ctgtcacgaa ggccagcacg tccaccttgc
     1081 gtcccagaaa ctgggagctc actggtctcc tgatatcgcc gtacataatg atatgctcgt
     1141 gaattttgat ccgcaggaag tgccccagca gtccccaaat attcttggac agcaggtcgg
     1201 cgatggtatg gccaatagtg gacccagcca cgctcagggt gtgcatgcct tcaggcccaa
     1261 agatgaacag gccgggaaac atcagccaac tagaatagta cacggcagtc atggccagca
     1321 ccagtttctt acagccgccg gcaggcatga tatagtaaga ctccacgtag atgattccag
     1381 cgttgtagaa tgtctgggtt ccatacacca ggccgataca gtaaaacagc cacttcaccc
     1441 acccagtggc cagagcggct gtcactccca tgcaaatagt tcccaggtcg gacaccagca
     1501 gagccattgt ccgcttattg tatgcctcac tcaggccggt gatgttagac agatgaatca
     1561 ggatcacggg acatgtcagc agccattcgg catatctcag ccatggggta atgttccctc
     1621 cgtacaggaa cagcatagca ggactggtga actcgaaata gatctccaga atcactttga
     1681 tcagctccac ggagcacacg tacacttcct cccatcccac gctggcctta taggcgtgcc
     1741 agccgtagaa agccagaaac agggcagaca ggaacacggc aaaccactgg aagcccctag
     1801 ctgtttcctg ctcccacagg gccccatagt agatgtagat ggagctgcac tgttcctcat
     1861 acccatgcac ggggtggtag ggtggattca cttgatatcc agctggggca tggtactcac
     1921 tgcctccggc agaatcgttt ccaggtgggg cgctgtgagc gtctggagca ggagctccgc
     1981 cgggaccggg gttttcttcc acgtctcctg cttgctttaa cagagagaag ttcgtggctc
     2041 cggagcccac ctcgttctcg tagcagaact tgtagaacct ggtggcctca gggaagtcgg
     2101 tggcgcagct gatgaagctg tcgatgctgc tcatgctcct catgtcgatc acgccctcgg
     2161 tcctggcagg cagaggggcc acagggctag gcatgctgct catctccagc tcctcaggag
     2221 gcttgctctg aggggccatc tccttggtgt tcaggatagg ctggctctgg gtacctccgc
     2281 tcccaccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     2341 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
     2401 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     2461 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     2521 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     2581 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     2641 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     2701 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     2761 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     2821 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca aatgaacttc agggtcagct
     2881 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     2941 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     3001 ccacgttgat gtcgatctgg tccaggggga tgtactcgcc ctcgctggtg atcctgctct
     3061 tggcggccgc cttgctcact gctactaccg ggcggtcggc ggcaccggta tcagctccag
     3121 ctgaaggttc aggtgcttct gtatcttgca gaatggctct tgacctgagc aggataaaac
     3181 cgaagcccac cttggcggtg acgtccagca ccatgaacag caaagtttcg attcccaggc
     3241 ccacaactcc ggcaccctcg gtgccgataa tccagatgat agggtaagca gtccacagga
     3301 ccagaaccag tgcggtgagg atgttgaagg ttgaggcaac ctctgggcct ctctccttag
     3361 cggctgaccg cagagaagtg gccaggacat acagaacgac gatcatacag atagttgaga
     3421 acagccacca agagtatctg gcaatggcag tatgtgacag tgttcctatc aggcctgtga
     3481 caatcatcag agcatcgacg ccaacgagag tgccgatggt cactctatca actttggcca
     3541 ggagtgccag gtgcagcagc aacagtggtg tagtgaacag ccactgtgcg tatcttgcgt
     3601 agtagatgtt cagcatctct ccacctactg gcacctctgt gaggccaata ccaaagaaca
     3661 ttgacagata agctgcactg gcaataccgc tgaccaggat tgtcactgcg tagtactccc
     3721 gtgcatcctt gtcagtctct ccccacgctc taaccaggaa gtagaaggtt ccgatcagca
     3781 tcagcagtgt accaattccg agccacagac tttcaggtct accatctccg aggaggtcat
     3841 aaccagcctg cagtgcgata gaaaccatgg tggcggagct agcataactt cgtataaagt
     3901 atcctatacg aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt
     3961 gcaaagaata acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta
     4021 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4081 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4141 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4201 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4261 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4321 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4381 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4441 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4501 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4561 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     4621 accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     4681 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4741 ctttatttgt aaccattata agctgcaata aactaaccac gtgcggaccg agcggccgca
     4801 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4861 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4921 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4981 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     5041 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5101 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5161 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5221 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5281 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5341 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5401 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5461 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5521 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5581 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5641 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5701 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5761 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5821 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5881 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5941 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6001 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6061 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6121 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6181 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6241 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6301 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6361 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6421 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6481 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6541 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6601 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6661 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6721 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6781 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6841 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7021 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7261 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7501 cggttcctgg ccttttgctg gccttttgct cacatgt
//
