LOCUS       sequence-205266-        6101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synOptopatch construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6101)
  AUTHORS   Fan LZ, Nehme R, Adam Y, Jung ES, Wu H, Eggan K, Arnold DB, Cohen AE
  TITLE     All-optical synaptic electrophysiology probes mechanism of 
            ketamine-induced disinhibition.
  JOURNAL   Nat Methods. 2018 Oct;15(10):823-831. doi: 
            10.1038/s41592-018-0142-8. Epub 2018 Oct 1.
  PUBMED    30275587
REFERENCE   2  (bases 1 to 6101)
  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"; doi: 
            "10.1038/s41592-018-0142-8"; journalName: "Nat Methods"; date: 
            "2018-10"; volume: "15"; issue: "10"; pages: "823-831"
COMMENT     Sequence Label: LZF1718: Cre-off hSyn-CheRiff-CFP (FAS)
FEATURES             Location/Qualifiers
     source          1..6101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      501..510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             507..1454
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="MGGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQ
                     CSSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKV
                     ILEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALL
                     VSDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLV
                     LAMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKI
                     HEHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     CDS             join(1524..1526,1527..2240)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1566..1587)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1827..1846)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2174..2195
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2312..2900
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2365..2385)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2783..2794)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2942..2959)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2948..3155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3184..3324
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3184..3313
                     /label=AAV2 ITR
     rep_origin      3399..3854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3486..3505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3696..3717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3871..3890)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3990..4012
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4050..4068)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4136..4240
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4241..4309,4310..5101)
                     /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(4459..4478)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5272..5860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5761..5780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5922..6051
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cacaacttcg tatatacctt
      481 tctatacgaa gttgtggcgc gccaccatgg gcggagctcc tgctccagac gctcacagcg
      541 ccccacctgg aaacgattct gccggaggca gtgagtacca tgccccagct ggatatcaag
      601 tgaatccacc ctaccacccc gtgcatgggt atgaggaaca gtgcagctcc atctacatct
      661 actatggggc cctgtgggag caggaaacag ctaggggctt ccagtggttt gccgtgttcc
      721 tgtctgccct gtttctggct ttctacggct ggcacgccta taaggccagc gtgggatggg
      781 aggaagtgta cgtgtgctcc gtggagctga tcaaagtgat tctggagatc tatttcgagt
      841 tcaccagtcc tgctatgctg ttcctgtacg gagggaacat taccccatgg ctgagatatg
      901 ccgaatggct gctgacatgt cccgtgatcc tgattcatct gtctaacatc accggcctga
      961 gtgaggcata caataagcgg acaatggctc tgctggtgtc cgacctggga actatttgca
     1021 tgggagtgac agccgctctg gccactgggt gggtgaagtg gctgttttac tgtatcggcc
     1081 tggtgtatgg aacccagaca ttctacaacg ctggaatcat ctacgtggag tcttactata
     1141 tcatgcctgc cggcggctgt aagaaactgg tgctggccat gactgccgtg tactattcta
     1201 gttggctgat gtttcccggc ctgttcatct ttgggcctga aggcatgcac accctgagcg
     1261 tggctgggtc cactattggc cataccatcg ccgacctgct gtccaagaat atttggggac
     1321 tgctggggca cttcctgcgg atcaaaattc acgagcatat cattatgtac ggcgatatca
     1381 ggagaccagt gagctcccag tttctgggac gcaaggtgga cgtgctggcc ttcgtgacag
     1441 aggaagataa agtggcggcc gccaagagca ggatcaccag cgagggcgag tacatccccc
     1501 tggaccagat cgacatcaac gtggtgagca agggcgagga gctgttcacc ggggtggtgc
     1561 ccatcctggt cgagctggac ggcgacgtaa acggccacag gttcagcgtg tccggcgagg
     1621 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1681 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc
     1741 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1801 tccaggagcg taccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1861 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1921 acggcaacat cctggggcac aagctggagt acaactacat cagccacaac gtctatatca
     1981 ccgccgacaa gcagaagaac ggcatcaagg cccacttcaa gatccgccac aacatcgagg
     2041 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2101 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2161 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2221 tggacgagct gtacaagtaa ttagctagct aaacaacttc gtatatacct ttctatacga
     2281 agttgtcgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2341 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2401 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2461 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2521 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2581 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2641 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2701 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     2761 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2821 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2881 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagctagagc tcgctgatca
     2941 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3001 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3061 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3121 gaggattggg aagacaatag caggcatgct ggggaggtaa ccacgtgcgg accgagcggc
     3181 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3241 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3301 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3361 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3421 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3481 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3541 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3601 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3661 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3721 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3781 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3841 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3901 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3961 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4021 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4081 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4141 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4201 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4261 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4321 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4381 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4441 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4501 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4561 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4621 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4681 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4741 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4801 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4861 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4921 gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact
     4981 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5041 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5101 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5881 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     5941 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     6001 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     6061 cgcgttgtac agtgtctaga ctgcagaggg ccctgcgtat g
//
