LOCUS       sequence-386295-        6387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NOPLight1 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6387)
  AUTHORS   Zhou X, Stine C, Prada PO, Fusca D, Assoumou K, Dernic J, Bhat MA, 
            Achanta AS, Johnson JC, Pasqualini AL, Jadhav S, Bauder CA, 
            Steuernagel L, Ravotto L, Benke D, Weber B, Suko A, Palmiter RD, 
            Stoeber M, Kloppenburg P, Bruning JC, Bruchas MR, Patriarchi T
  TITLE     Development of a genetically encoded sensor for probing endogenous 
            nociceptin opioid peptide release.
  JOURNAL   Nat Commun. 2024 Jun 25;15(1):5353. doi: 10.1038/s41467-024-49712-0.
  PUBMED    38918403
REFERENCE   2  (bases 1 to 6387)
  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 
            Commun."; date: "2024-06-25"; volume: "15(1):5353. doi"; pages: " 
            10.1038/s41467-024-49712-0"
COMMENT     Sequence Label: pAAV_hSynapsin1_NOPLight1
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          635..767
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     750..769
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2766..3354
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2819..2839)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3237..3248)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3372..3596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3639..3779
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3639..3768
                     /label=AAV2 ITR
     rep_origin      3854..4309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3941..3960)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4151..4172
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4326..4345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4445..4467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4505..4523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4591..4695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4696..4764,4765..5556)
                     /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(4914..4933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5727..6315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6216..6235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6377..6387,1..119)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgtgtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      601 tgagagcgca gtcgagatta attaactcga gcaggtaagt atcaaggtta caagacaggt
      661 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
      721 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg ctagcgttta
      781 aacggtaccg gatccggtac ctgcgcgcca tgaagacgat catcgccctg agctacatct
      841 tctgcctggt gttcgccgac tacaaggacg atgatgacgc catggagcca ctttttccgg
      901 ctcccttttg ggaagttatc tatgggtctc acctccaggg gaatttgagt cttcttagcc
      961 cgaaccattc tctgctgcct ccacaccttt tgctgaacgc gtcacacggt gcctttcttc
     1021 cgctcggtct taaggtaacc attgttggac tctatcttgc tgtgtgcgtt ggaggcttgc
     1081 tggggaattg tcttgtcatg tatgtgatct tgcgacatac caagatgaag actgctacaa
     1141 atatatacat atttaatctg gcacttgctg atactcttgt gttgctcacg ctgccctttc
     1201 aagggactga catactcctt gggttctggc cgttcggtaa tgcactctgc aaaaccgtga
     1261 tcgcaattga ttattacaat atgtttacgt caacgtttac tcttaccgct atgagtgtgg
     1321 atcggtacgt cgctatttgt caccctaaac gagcgctgga tgttcgaacc agttcaaaag
     1381 cacaggccgt caatgtagct atctgggcac ttgcaagtgt ggtcggtgtg cctgtggcga
     1441 ttatggggtc tgcgcaagta gaagacgaag agattgagtg cttggttgag atccctacac
     1501 cccaggacta ttggggaccc gtattcgcaa tttgtatttt tctcttttct tttattgtcc
     1561 ccgttctggt aatttctgtc tgttatagtc tcatgatacg ccgcctgagg ggggtgcgca
     1621 tagcccagaa gaatagctct ctcatcaacg tatatataaa agccgataaa caaaagaatg
     1681 gcattaaggc gaatttcaag attaggcaca atatagaaga cggaggtgtt caacttgcgt
     1741 accattatca gcagaatacc ccgattgggg atggtcctgt gcttcttccc gacaatcact
     1801 atctgagcgt acagagcaaa ctctccaagg acccgaacga gaaacgggat cacatggtgc
     1861 tgttggaatt tgtaacggca gcggggatta ctctgggcat ggacgaattg tataaaggag
     1921 gcacgggggg ttcaatggtt agtaaaggag aggaactttt tacaggcgtc gtcccgattt
     1981 tggtagaatt ggatggcgat gtaaacgggc ataaatttag tgtatccggc gagggagagg
     2041 gagatgctac ctacgggaag ctcaccctga aattcatctg cacgacaggt aaattgcctg
     2101 ttccgtggcc gactttggtg acgactctga cctatggtgt ccagtgtttc tctcggtatc
     2161 cagaccatat gaagcagcat gacttcttca agagcgcgat gccagagggt tacattcaag
     2221 aaaggactat attctttaaa gatgacggca attacaagac tcgcgctgaa gttaaattcg
     2281 aaggtgatac attggtaaac cgaatagaac ttaagggtat agacttcaaa gaagatggaa
     2341 atatactggg ccacaaactt gagtacaaca accacgatca gaaggacaga aatttgcgac
     2401 ggatcacgcg gctcgtgctc gttgtggttg ctgtttttgt tggttgttgg accccggttc
     2461 aggtctttgt gttggcccaa ggtctcgggg tacaacccag ctcagaaaca gctgtcgcga
     2521 tattgaggtt ttgtacggcg cttggatacg tcaactcatg cttgaaccct atactgtacg
     2581 ctttcctcga cgaaaacttt aaaaggtgtt ttcgagactt ctgcttcccg ttgaagatgc
     2641 gaatggagag acaggccact gcgagagtgc ggaatacggt acaggacccg gcggccctcc
     2701 gagacatcga cggaatgaac aagcctgtat aatgtacagc ggccgcgact ctaaagctta
     2761 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2821 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2881 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2941 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3001 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3061 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3121 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3181 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3241 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3301 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgctctcga
     3361 gatacgtacg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3421 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3481 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3541 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3601 tgaggcggaa agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     3661 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3721 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     3781 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3841 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4201 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4321 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4381 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4441 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4501 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4561 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4621 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4681 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4741 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4801 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4861 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4921 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4981 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5041 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5101 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5161 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5221 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5281 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5341 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5401 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5461 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5521 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6361 ccttttgctc acatgtcctg caggcag
//
