LOCUS       sequence-390701-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses optimized PdCO in frame with mScarlet under control of 
            human synapsin1 promotor after Cre-dependent recombination..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  AUTHORS   Wietek J, Nozownik A, Pulin M, Saraf-Sinik I, Matosevich N, 
            Gowrishankar R, Gat A, Malan D, Brown BJ, Dine J, Imambocus BN, Levy
            R, Sauter K, Litvin A, Regev N, Subramaniam S, Abrera K, Summarli D,
            Goren EM, Mizrachi G, Bitton E, Benjamin A, Copits BA, Sasse P, Rost
            BR, Schmitz D, Bruchas MR, Soba P, Oren-Suissa M, Nir Y, Wiegert JS,
            Yizhar O
  TITLE     A bistable inhibitory optoGPCR for multiplexed optogenetic control 
            of neural circuits.
  JOURNAL   Nat Methods. 2024 May 29. doi: 10.1038/s41592-024-02285-8.
  PUBMED    38811857
REFERENCE   2  (bases 1 to 6506)
  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: "Nat 
            Methods. 2024 May 29. doi: 10.1038/s41592-024-02285-8."
COMMENT     Sequence Label: pAAV-hSyn-SIO-PdCO-mScarlet-WPRE
FEATURES             Location/Qualifiers
     source          1..6506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      26..614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     515..534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   676..805
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        850..1297
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(1311..1344)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1378..2073)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1653..1672)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1922..1940
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3246..3255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3262..3295
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3308..3896
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3361..3381)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3779..3790)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3928..4404
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4127..4146)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4444..4584
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4444..4573
                     /label=AAV2 ITR
     rep_origin      4659..5114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4746..4765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4956..4977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5131..5150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5250..5272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5310..5328)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5396..5500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5501..5569,5570..6361)
                     /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(5719..5738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
       61 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      121 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      361 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      421 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      481 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      541 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      601 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      661 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
      721 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      781 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc
      841 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      901 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      961 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
     1021 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
     1081 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
     1141 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
     1201 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
     1261 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc taccgttcgt
     1321 atagcataca ttatacgaag ttatgaattc tagttacacc tcgttctcgt agcagaactt
     1381 gtacagctcg tccatgccgc cggtggagtg gcggccctcg gagcgttcgt actgttccac
     1441 cacggtgtag tcctcgttgt gggaggtgat gtccaacttg cggtcgacgt tgtaggcgcc
     1501 gggcatctgc acgggcttct tggccttgta ggtggtcttg aagtccgcca ggtagcgtcc
     1561 gccgtccttc aggcgcaggg ccatcttaat gtcgcccttc agcacgccgt cctcggggta
     1621 caaccgctcg gtggacgctt cccagcccat tgtcttcttc tgcattacgg ggccgtcagg
     1681 agggaagttg gtgccgcgga gcttcacctt gtagatcagg gtgccgtcct ccagggaggt
     1741 gtcctgggtc acggtcacgg cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa
     1801 gccctcgggg aaggactgct tatagtagtc ggggatgtcg gcggggtgct tggtgaaggc
     1861 cctggagccg tacatgaact gaggggacag gatgtcccag gagaagggca gggggccacc
     1921 cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc
     1981 ctcgatctcg aactcgtggc cgttcatgga gccctccatg tgcaccttga accgcatgaa
     2041 ctccttgatc actgcctcgc ccttgctcac catcaccacg ttgatgtcga tctggtccag
     2101 ggggatgtac tcgccctcgc tggtgatcct gctcttaccg gttggatctc gggctcgcgg
     2161 agcaggagca acctgggaag tctcggtcgt cgcggcgact tgattcgtgg ctgtaccacc
     2221 agttgctctc tcactctcgg agttcaggtc ctctactccc attccgagaa gaactttaag
     2281 gctccgcctg aattgtttgt tcatgaacac atagataaga gggttataca tcgtagaaga
     2341 ctttgcaaac atggcaggaa tcactgtggc cactggatgg agcaggtctt tagggccgaa
     2401 gctagcgagc atgcagataa cagcgtaggg actccaggca agcatgaaca ggctaaccat
     2461 cagcagcgtc atgcgcagta cttcgcgttc tgctttggca acagaaccgc ctgttttcgc
     2521 taccttccga accttatcgt agatcagccc gtatgacacg acgataatga gaacagggac
     2581 aagaaagcag aagatgaaaa tagcaaacac gtaggactta tcggatgttt catcggaaag
     2641 ccagtcgata ctacaccaag tggccaaacc ttccggtatg tagtgattcc acccaaggag
     2701 aggcgcgagt gtgaatgcca gggagtagat ccaaatgaaa gccatcgccg ccacgcccat
     2761 ccgcgttgtc aaacctgtga aggagccgag acgcagcacg gccagacaac gaacaaaagc
     2821 aataacgctc agggtcatca gtgccgcgag cccgccgagg tatgttatga agccgtacca
     2881 tttgcagaaa ccgtcttctt ctcgcaacag ggttttgcga gctgtaataa tgaacttaaa
     2941 tgcggtgccg aaaactgcaa ccaacagatc cccgagagcg agacttacat acaggaagtt
     3001 attggggctc ctcaactttc tgtctttgat aaagaggacc aacaccaaaa cgttgagact
     3061 gactcccaga caagcaatga agaaaaggta agctgcacaa atgttgtaat cggtggctgt
     3121 gaaggcattg tagatgtaat gatttccgcc gtccgtttcc gcattaaggt tttggaaata
     3181 cgtgctgttg attggggtat ccatcaactc ggtgacagga tttggaatcg tcaagttctc
     3241 cccgtccatg gtggcggatc ctaccgttcg tataatgtat gctatacgaa gttataagct
     3301 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3361 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3421 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3481 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3541 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3601 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3661 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3721 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3781 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3841 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3901 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     3961 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     4021 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     4081 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     4141 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     4201 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     4261 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     4321 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     4381 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     4441 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4501 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4561 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4621 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4681 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4741 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4801 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4861 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4921 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4981 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     5041 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5101 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5161 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5221 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5281 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5341 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5401 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5461 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5521 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5581 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5641 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5701 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5761 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5821 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5881 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5941 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6001 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6061 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6121 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6181 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6241 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6301 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6361 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6421 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6481 gttttcgttc cactgagcgt cagacc
//
