LOCUS       sequence-353305-        7628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-on bicistronic expression of QuasAr6b-based Optopatch under an 
            neuronal promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7628)
  AUTHORS   Tian H, Davis HC, Wong-Campos JD, Park P, Fan LZ, Gmeiner B, Begum 
            S, Werley CA, Borja GB, Upadhyay H, Shah H, Jacques J, Qi Y, Parot 
            V, Deisseroth K, Cohen AE
  TITLE     Video-based pooled screening yields improved far-red genetically 
            encoded voltage indicators.
  JOURNAL   Nat Methods. 2023 Jul;20(7):1082-1094. doi: 
            10.1038/s41592-022-01743-5. Epub 2023 Jan 9.
  PUBMED    36624211
REFERENCE   2  (bases 1 to 7628)
  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/s41592-022-01743-5"; journalName: "Nat Methods"; date: 
            "2023-07"; volume: "20"; issue: "7"; pages: "1082-1094"
COMMENT     Sequence Label: 
            HT107_pAAV_hSyn-DiO-SomQuasAr6a_EGFP-P2A-somCheRiff_HA
FEATURES             Location/Qualifiers
     source          1..7628
                     /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     complement(2319..2340)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     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(4651..4668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4657..4864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4891..5031
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4891..5020
                     /label=AAV2 ITR
     rep_origin      5106..5561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5193..5212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5403..5424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5578..5597)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5697..5719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5757..5775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5843..5947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5948..6016,6017..6808)
                     /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(6166..6185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6979..7567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7468..7487
                     /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 caggaccatg
     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 aatgcttctt 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 caggcaccaa
     3421 gagtatctgg caatggcagt atgtgacagt gtccctatca ggcctgtgac aatcatcaga
     3481 gcatcgccgc caacgagagt gccgatggtc actctatcaa ctttggccag gagtgccagg
     3541 tgcagcagca acagtggtgt agtcagcagc cactgtgcgt atcttgcgta gtagatgttc
     3601 aggatctctc cacctactgg cacctctgtg aggccaatac caaagaacat tgacagataa
     3661 gctgcactgg caataccgct gaccaggatt gtcactgcgt agtactcccg tgcatccttg
     3721 tcagtctctc cgcccgctct 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 ccgagcgctg
     4621 ctcgagagat ctctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     4681 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4741 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4801 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4861 gggataacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4921 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4981 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5041 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5101 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5161 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     5221 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5281 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5341 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5401 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     5461 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     5521 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5581 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5641 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5701 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5761 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5821 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5881 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5941 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6001 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6061 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6121 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6181 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6241 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6301 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6361 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6421 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6481 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6541 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6601 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6661 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6721 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6781 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6841 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6901 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6961 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7021 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7081 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7141 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7201 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7261 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7321 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7381 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7441 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7501 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7561 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7621 tcacatgt
//
