LOCUS       sequence-247758-        7526 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 7526)
  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 7526)
  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..7526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(984..1010)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1026..1970)
                     /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(1971..2027)
                     /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(2277..2987,2988..2990))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(2277..2987,2988..2990))
                     /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     2668..2687
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2927..2948
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3854..3863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3873..3906
                     /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    3957..3990
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4015..4603
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4068..4088)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4486..4497)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4657..4676)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4711..4730
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4789..4929
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4789..4918
                     /label=AAV2 ITR
     rep_origin      5004..5459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5091..5110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5301..5322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5476..5495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5595..5617
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5655..5673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5741..5845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5846..5914,5915..6706)
                     /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(6064..6083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6877..7465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7366..7385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccttaga atcgagtagc ctctggaaaa tcggtggcgc atgaaatgaa gctgtcgata
      841 ctggacatag atcgcatgtc tataacgcct tcagtccgtg ctggaagggg tgcgacaggg
      901 ctgggcatgg atgacatctc caattcctcg ggtggttttg attgtggggc catttcctta
      961 gtgttaagta taggctgact ctgagcgtaa tctggaacat cgtatgggta gctcttggcg
     1021 gccgccactt tatcttcctc tgtcacgaag gccagcacgt ccaccttgcg tcccagaaac
     1081 tgggagctca ctggtctcct gatatcgccg tacataatga tatgctcgtg aattttgatc
     1141 cgcaggaagt gccccagcag tccccaaata ttcttggaca gcaggtcggc gatggtatgg
     1201 ccaatagtgg acccagccac gctcagggtg tgcatgcctt caggcccaaa gatgaacagg
     1261 ccgggaaaca tcagccaact agaatagtac acggcagtca tggccagcac cagtttctta
     1321 cagccgccgg caggcatgat atagtaagac tccacgtaga tgattccagc gttgtagaat
     1381 gtctgggttc catacaccag gccgatacag taaaacagcc acttcaccca cccagtggcc
     1441 agagcggctg tcactcccat gcaaatagtt cccaggtcgg acaccagcag agccattgtc
     1501 cgcttattgt atgcctcact caggccggtg atgttagaca gatgaatcag gatcacggga
     1561 catgtcagca gccattcggc atatctcagc catggggtaa tgttccctcc gtacaggaac
     1621 agcatagcag gactggtgaa ctcgaaatag atctccagaa tcactttgat cagctccacg
     1681 gagcacacgt acacttcctc ccatcccacg ctggccttat aggcgtgcca gccgtagaaa
     1741 gccagaaaca gggcagacag gaacacggca aaccactgga agcccctagc tgtttcctgc
     1801 tcccacaggg ccccatagta gatgtagatg gagctgcact gttcctcata cccatgcacg
     1861 gggtggtagg gtggattcac ttgatatcca gctggggcat ggtactcact gcctccggca
     1921 gaatcgtttc caggtggggc gctgtgagcg tctggagcag gagctccgcc gggaccgggg
     1981 ttttcttcca cgtctcctgc ttgctttaac agagagaagt tcgtggctcc ggagcccacc
     2041 tcgttctcgt agcagaactt gtagaacctg gtggcctcag ggaagtcggt ggcgcagctg
     2101 atgaagctgt cgatgctgct catgctcctc atgtcgatca cgccctcggt cctggcaggc
     2161 agaggggcca cagggctagg catgctgctc atctccagct cctcaggagg cttgctctga
     2221 ggggccatct ccttggtgtt caggataggc tggctctggg tacctccgct cccacccttg
     2281 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag taggaccatg
     2341 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     2401 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     2461 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     2521 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     2581 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     2641 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     2701 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     2761 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     2821 cagggcacgg gcagcttgcc ggtggtgcaa atgaacttca gggtcagctt gccgtaggtg
     2881 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     2941 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac cacgttgatg
     3001 tcgatctggt ccagggggat gtactcgccc tcgctggtga tcctgctctt ggcggccgcc
     3061 ttgctcactg ctactaccgg gcggtcggcg gcaccggtat cagctccagc tgaaggttca
     3121 ggtgcttctg tatcttgcag aatggctctt gacctgagca ggataaaacc gaagcccacc
     3181 ttggcggtga cgtccagcac catgaacagc aaagtttcga ttcccaggcc cacaactccg
     3241 gcaccctcgg tgccgataat ccagatgata gggtaagcag tccacaggac cagaaccagt
     3301 gcggtgagga tgttgaaggt tgaggcaacc tctgggcctc tctccttagc ggctgaccgc
     3361 agagaagtgg ccaggacata cagaacgacg atcatacaga tagttgagaa cagccaccaa
     3421 gagtatctgg caatggcagt atgtgacagt gttcctatca ggcctgtgac aatcatcaga
     3481 gcatcgacgc caacgagagt gccgatggtc actctatcaa ctttggccag gagtgccagg
     3541 tgcagcagca acagtggtgt agtgaacagc cactgtgcgt atcttgcgta gtagatgttc
     3601 agcatctctc cacctactgg cacctctgtg aggccaatac caaagaacat tgacagataa
     3661 gctgcactgg caataccgct gaccaggatt gtcactgcgt agtactcccg tgcatccttg
     3721 tcagtctctc cccacgctct aaccaggaag tagaaggttc cgatcagcat cagcagtgta
     3781 ccaattccga gccacagact ttcaggtcta ccatctccga ggaggtcata accagcctgc
     3841 agtgcgatag aaaccatggt ggcggagcta gcataacttc gtataaagta tcctatacga
     3901 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     3961 cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa
     4021 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4081 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4141 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4201 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4261 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4321 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4381 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     4441 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4501 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4561 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc
     4621 gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     4681 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4741 accattataa gctgcaataa actaaccacg tgcggaccga gcggccgcag gaacccctag
     4801 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4861 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4921 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4981 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5041 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     5101 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5161 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5221 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5281 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     5341 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     5401 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     5461 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5521 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5581 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5641 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5701 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5761 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5821 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5881 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5941 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6001 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6061 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6121 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6181 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6241 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6301 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6361 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6421 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6481 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6541 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6601 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6661 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6721 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6781 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6841 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6901 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6961 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7021 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7081 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7141 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7201 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7261 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7321 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7381 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7441 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7501 cttttgctgg ccttttgctc acatgt
//
