LOCUS       sequence-196842-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of eNpHR 3.0 fused to EYFP driven by humanized 
            Synapsin promoter for optogenetic inhibition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  TITLE     eNpHR3.0-EYFP
REFERENCE   2  (bases 1 to 6242)
  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"
COMMENT     Sequence Label: pAAV-hSyn-eNpHR 3.0-EYFP
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   52..181
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        226..673
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1655..1657,1658..2368)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1697..1718)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1958..1977)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2305..2326
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2420..3008
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2473..2493)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2891..2902)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3040..3516
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3239..3258)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3556..3696
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3556..3685
                     /label=AAV2 ITR
     rep_origin      3771..4226
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3858..3877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4068..4089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4243..4262)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4362..4384
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4422..4440)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4508..4612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4613..4681,4682..5473)
                     /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(4831..4850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5644..6232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6133..6152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
       61 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
      121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      181 tgcggccgca cgcgtgtgtc tagactgcag agggccctgc gtatgagtgc aagtgggttt
      241 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      301 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      361 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      421 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      481 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      541 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      601 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      661 cctgagagcg cagtcgagaa ggtaccggat ccccgggtac cggtaggcca ccatgacaga
      721 gaccctgcct cccgtgaccg agagtgccgt ggcccttcaa gccgaggtta cccaaaggga
      781 gttgttcgag ttcgtgctga acgacccttt gcttgcaagc agtctctata tcaacatcgc
      841 acttgcagga ctgagtatac tgctgttcgt ttttatgacc cgaggactcg atgatccacg
      901 ggcaaaactt attgctgtgt caaccatcct tgtgcctgtc gtcagcattg cctcctacac
      961 tggattggcg agcggcctga caatttccgt tcttgaaatg ccagcgggcc attttgcaga
     1021 aggcagctca gtgatgctgg gaggagaaga ggtagatggt gtagtcacca tgtggggacg
     1081 gtatctcacc tgggcacttt ccacgcccat gattctcctc gctctgggtc tcctggccgg
     1141 aagcaatgct acaaagctct tcacagctat cactttcgat atcgctatgt gcgtgactgg
     1201 ccttgccgcg gccctgacta cctcctccca cctcatgaga tggttctggt acgctatcag
     1261 ttgtgcatgc tttctggtgg tcttgtatat cctgctggtg gagtgggcac aggacgccaa
     1321 agccgcggga accgctgaca tgttcaatac cctgaagctg ttgacagtag tgatgtggct
     1381 ggggtatcca attgtgtggg ctcttggagt cgagggtatc gcggtgttgc ccgttggggt
     1441 gacgagctgg ggatattctt tcctggatat cgtggcaaag tacattttcg cattcttgct
     1501 cctgaactat ctgacgtcaa acgaatctgt cgtgtccggc agcattttgg atgttccatc
     1561 tgcttctggg accccggctg atgatgcggc cgccaagagc aggatcacca gcgagggcga
     1621 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac
     1681 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1741 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1801 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
     1861 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1921 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1981 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2041 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2101 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2161 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2221 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     2281 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2341 cactctcggc atggacgagc tgtacaagtt ctgctacgag aacgaggtgt aatgagaatt
     2401 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2461 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2521 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2581 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2641 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2701 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2761 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2821 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     2881 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2941 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3001 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     3061 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3121 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3181 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3241 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3301 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     3361 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3421 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3481 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     3541 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3601 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3661 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3721 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3841 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3961 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4021 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4081 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     4141 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4201 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4261 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4321 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4381 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4441 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4501 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4561 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4621 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4681 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4741 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4801 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4861 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4921 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4981 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5041 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5101 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5161 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5221 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5281 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5341 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5401 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5461 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5521 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5581 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5641 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5701 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5761 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5821 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5881 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5941 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6001 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6061 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6241 ca
//
