LOCUS       sequence-390699-        5805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses optimized PdCO in frame with EGFP under control of human 
            synapsin1 promotor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  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 5805)
  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-PdCO-EGFP-WPRE
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   193..322
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        367..814
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      834..843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2016..2018,2019..2021,2022..2732)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2016..2018,2019..2021,2022..2732)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2061..2082)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2322..2341)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2669..2690
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2772..3175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    3182..3230
                     /note="synthetic polyadenylation signal"
     repeat_region   3260..3400
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3260..3389
                     /label=AAV2 ITR
     rep_origin      3475..3930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3562..3581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3772..3793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3947..3966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4066..4088
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4126..4144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4212..4316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4317..4385,4386..5177)
                     /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(4535..4554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5348..5805,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggttttg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
      241 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      301 aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg
      361 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac
      421 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
      481 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
      541 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
      601 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
      661 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
      721 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
      781 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tccgccacca
      841 tggacgggga gaacttgacg attccaaatc ctgtcaccga gttgatggat accccaatca
      901 acagcacgta tttccaaaac cttaatgcgg aaacggacgg cggaaatcat tacatctaca
      961 atgccttcac agccaccgat tacaacattt gtgcagctta ccttttcttc attgcttgtc
     1021 tgggagtcag tctcaacgtt ttggtgttgg tcctctttat caaagacaga aagttgagga
     1081 gccccaataa cttcctgtat gtaagtctcg ctctcgggga tctgttggtt gcagttttcg
     1141 gcaccgcatt taagttcatt attacagctc gcaaaaccct gttgcgagaa gaagacggtt
     1201 tctgcaaatg gtacggcttc ataacatacc tcggcgggct cgcggcactg atgaccctga
     1261 gcgttattgc ttttgttcgt tgtctggccg tgctgcgtct cggctccttc acaggtttga
     1321 caacgcggat gggcgtggcg gcgatggctt tcatttggat ctactccctg gcattcacac
     1381 tcgcgcctct ccttgggtgg aatcactaca taccggaagg tttggccact tggtgtagta
     1441 tcgactggct ttccgatgaa acatccgata agtcctacgt gtttgctatt ttcatcttct
     1501 gctttcttgt ccctgttctc attatcgtcg tgtcatacgg gctgatctac gataaggttc
     1561 ggaaggtagc gaaaacaggc ggttctgttg ccaaagcaga acgcgaagta ctgcgcatga
     1621 cgctgctgat ggttagcctg ttcatgcttg cctggagtcc ctacgctgtt atctgcatgc
     1681 tcgctagctt cggccctaaa gacctgctcc atccagtggc cacagtgatt cctgccatgt
     1741 ttgcaaagtc ttctacgatg tataaccctc ttatctatgt gttcatgaac aaacaattca
     1801 ggcggagcct taaagttctt ctcggaatgg gagtagagga cctgaactcc gagagtgaga
     1861 gagcaactgg tggtacagcc acgaatcaag tcgccgcgac gaccgagact tcccaggttg
     1921 ctcctgctcc gcgagcccga gatccaaccg gtaagagcag gatcaccagc gagggcgagt
     1981 acatccccct ggaccagatc gacatcaacg tggtgatggt gagcaagggc gaggagctgt
     2041 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2101 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcattt
     2161 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2221 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2281 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2341 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2401 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2461 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2521 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2581 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2641 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2701 ggatcactct cggcatggac gagctgtaca agttctgcta cgagaacgag gtgtaactag
     2761 aattcaagct taatcaacct ctggattaca aaatttgtga aagattgact gatattctta
     2821 actatgttgc tccttttacg ctgtgtggat atgctgcttt aatgcctctg tatcatgcta
     2881 ttgcttcccg tatggctttc gttttctcct ccttgtataa atcctggttg ctgtctcttt
     2941 atgaggagtt gtggcccgtt gtccgtcaac gtggcgtggt gtgctctgtg tttgctgacg
     3001 caacccccac tggctggggc attgccacca cctgtcaact cctttctggg actttcgctt
     3061 tccccctccc gatcgccacg gcagaactca tcgccgcctg ccttgcccgc tgctggacag
     3121 gggctaggtt gctgggcact gataattccg tggtgttgtc ggggaagctg acgtcagcgc
     3181 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg ggtaacccac
     3241 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3301 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3361 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3421 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3481 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3541 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3601 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3661 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3721 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3781 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3841 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3901 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3961 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4021 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4081 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4141 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4201 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4261 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4321 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4381 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4441 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4501 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4561 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4621 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4681 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4741 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4801 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4861 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4921 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4981 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5041 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5101 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5161 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5221 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5281 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5341 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5401 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5461 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5521 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5581 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5641 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5701 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5761 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcg
//
