LOCUS       sequence-268574-        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 7508)
  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 7508)
  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: LZF1826 pAAV_hSyn-DiO-SomArchon-EGFP-P2A-stGtACR2
FEATURES             Location/Qualifiers
     source          1..7508
                     /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(1953..2009)
                     /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(2259..2969,2970..2972))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(2259..2969,2970..2972))
                     /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     2650..2669
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2909..2930
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3836..3845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3855..3888
                     /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    3939..3972
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3997..4585
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4050..4070)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4468..4479)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4639..4658)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4693..4712
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4771..4911
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4771..4900
                     /label=AAV2 ITR
     rep_origin      4986..5441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5073..5092)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5283..5304
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5458..5477)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5577..5599
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5637..5655)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5723..5827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5828..5896,5897..6688)
                     /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(6046..6065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6859..7447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7348..7367
                     /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 caccacgttg
     1021 atgtcgatct ggtccagggg gatgtactcg ccctcgctgg tgatcctgct cttggcggcc
     1081 cttccaaaca ttctgatatt gacttcgtcg ttaggagggg cattcttctc gtgattcagg
     1141 ccaaaaggat cgtcggcttc gggtttctcc tgccctggca gacatgcggg gtcccacttc
     1201 ccttccagca ctccccaggc agtgctccac aggatcactc catatgtatt tttgcacagg
     1261 ccgtcagcca gcaggtacag aatggcgctc acgttctcgt ggatcacgca gactccggtg
     1321 ggtgaaatca gccacacgat tgggaacata gtccaagcaa taaagaagat caggcgcagc
     1381 atcttgatcc tcaccaccac cttctgtgcc agaggtgtcc cgacagattc aaagccatac
     1441 agccccactt gaaagattgt gaagaccaca ctgtactcga acaccaggca ggtggcccca
     1501 aagaaaaaga acagccactt catatactct gcgggagaaa ttgtagcggt gattcccatg
     1561 accactctaa tgatggaagc ggcctgtgca atgggattca gtgggatact tttgtacttc
     1621 accagagcca tgttagaaat ctgtcccagc atgattggac aggtgcacag ccaggaggcc
     1681 attcggctcc aaggaaagaa tctgccgtcc ctcatctgca gggttccgtt gccagtactg
     1741 gccagtgcat aagtgataga ttcgacaaag ggcaggtaca cggactccca agggaccttg
     1801 ggagccctgc taatcatgaa gattgtctgg cagcctgaga ccacggccca caccagctgc
     1861 agcagtgaga cgaaggtgga gtcgattctg gacatattgt agcactcggt gtgagttgat
     1921 gcccattccc catagacgac ctgtgatgcc atgggaccgg ggttttcttc cacgtctcct
     1981 gcttgcttta acagagagaa gttcgtggct ccggagccca cctcgttctc gtagcagaac
     2041 ttgtagaacc tggtggcctc agggaagtcg gtggcgcagc tgatgaagct gtcgatgctg
     2101 ctcatgctcc tcatgtcgat cacgccctcg gtcctggcag gcagaggggc cacagggcta
     2161 ggcatgctgc tcatctccag ctcctcagga ggcttgctct gaggggccat ctccttggtg
     2221 ttcaggatag gctggctctg ggtacctccg ctcccaccct tgtacagctc gtccatgccg
     2281 agagtgatcc cggcggcggt cacgaactcc agtaggacca tgtgatcgcg cttctcgttg
     2341 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     2401 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     2461 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
     2521 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     2581 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     2641 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     2701 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     2761 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
     2821 ccggtggtgc aaatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     2881 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     2941 accccggtga acagctcctc gcccttgctc accacgttga tgtcgatctg gtccaggggg
     3001 atgtactcgc cctcgctggt gatcctgctc ttggcggccg ccttgctcac tgctactacc
     3061 gggcggtcgg cggcaccggt atcagctcca gctgaaggtt caggtgcttc tgtatcttgc
     3121 agaatggctc ttgacctgag caggataaaa ccgaagccca ccttggcggt gacgtccagc
     3181 accatgaaca gcaaagtttc gattcccagg cccacaactc cggcaccctc ggtgccgata
     3241 atccagatga tagggtaagc agtccacagg accagaacca gtgcggtgag gatgttgaag
     3301 gttgaggcaa cctctgggcc tctctcctta gcggctgacc gcagagaagt ggccaggaca
     3361 tacagaacga cgatcataca gatagttgag aacagccacc aagagtatct ggcaatggca
     3421 gtatgtgaca gtgttcctat caggcctgtg acaatcatca gagcatcgac gccaacgaga
     3481 gtgccgatgg tcactctatc aactttggcc aggagtgcca ggtgcagcag caacagtggt
     3541 gtagtgaaca gccactgtgc gtatcttgcg tagtagatgt tcagcatctc tccacctact
     3601 ggcacctctg tgaggccaat accaaagaac attgacagat aagctgcact ggcaataccg
     3661 ctgaccagga ttgtcactgc gtagtactcc cgtgcatcct tgtcagtctc tccccacgct
     3721 ctaaccagga agtagaaggt tccgatcagc atcagcagtg taccaattcc gagccacaga
     3781 ctttcaggtc taccatctcc gaggaggtca taaccagcct gcagtgcgat agaaaccatg
     3841 gtggcggagc tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc
     3901 agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct
     3961 atacgaagtt atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     4021 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4081 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4141 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4201 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4261 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4321 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4381 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     4441 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4501 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4561 atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc gcttcgagca
     4621 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4681 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4741 aaactaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4801 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4861 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     4921 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     4981 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5041 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     5101 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5161 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5221 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5281 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     5341 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     5401 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5461 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5521 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5581 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5641 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5701 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5761 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5821 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5881 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5941 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6001 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6061 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6121 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6181 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6241 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6301 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6361 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6421 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6481 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6541 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6601 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6661 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6721 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6781 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6961 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7441 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7501 tcacatgt
//
