LOCUS       sequence-386290-        5910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NOPLight1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5910)
  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 5910)
  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: pCMV_NOPlight1
FEATURES             Location/Qualifiers
     source          1..5910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2334..2455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2371..2390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2425..2444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2462..2917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2599..2620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2811..2830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2944..3048
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3050..3407
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3050..3241
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3073..3093)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3258..3393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3442..4236
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3442..4236
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3496..3515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4106..4125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4424..4443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4468..4515
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4844..5432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5333..5352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5607..5910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttggatccg gtacctgcgc gccatgaaga cgatcatcgc
      301 cctgagctac atcttctgcc tggtgttcgc cgactacaag gacgatgatg acgccatgga
      361 gccacttttt ccggctccct tttgggaagt tatctatggg tctcacctcc aggggaattt
      421 gagtcttctt agcccgaacc attctctgct gcctccacac cttttgctga acgcgtcaca
      481 cggtgccttt cttccgctcg gtcttaaggt aaccattgtt ggactctatc ttgctgtgtg
      541 cgttggaggc ttgctgggga attgtcttgt catgtatgtg atcttgcgac ataccaagat
      601 gaagactgct acaaatatat acatatttaa tctggcactt gctgatactc ttgtgttgct
      661 cacgctgccc tttcaaggga ctgacatact ccttgggttc tggccgttcg gtaatgcact
      721 ctgcaaaacc gtgatcgcaa ttgattatta caatatgttt acgtcaacgt ttactcttac
      781 cgctatgagt gtggatcggt acgtcgctat ttgtcaccct aaacgagcgc tggatgttcg
      841 aaccagttca aaagcacagg ccgtcaatgt agctatctgg gcacttgcaa gtgtggtcgg
      901 tgtgcctgtg gcgattatgg ggtctgcgca agtagaagac gaagagattg agtgcttggt
      961 tgagatccct acaccccagg actattgggg acccgtattc gcaatttgta tttttctctt
     1021 ttcttttatt gtccccgttc tggtaatttc tgtctgttat agtctcatga tacgccgcct
     1081 gaggggggtg cgcatagccc agaagaatag ctctctcatc aacgtatata taaaagccga
     1141 taaacaaaag aatggcatta aggcgaattt caagattagg cacaatatag aagacggagg
     1201 tgttcaactt gcgtaccatt atcagcagaa taccccgatt ggggatggtc ctgtgcttct
     1261 tcccgacaat cactatctga gcgtacagag caaactctcc aaggacccga acgagaaacg
     1321 ggatcacatg gtgctgttgg aatttgtaac ggcagcgggg attactctgg gcatggacga
     1381 attgtataaa ggaggcacgg ggggttcaat ggttagtaaa ggagaggaac tttttacagg
     1441 cgtcgtcccg attttggtag aattggatgg cgatgtaaac gggcataaat ttagtgtatc
     1501 cggcgaggga gagggagatg ctacctacgg gaagctcacc ctgaaattca tctgcacgac
     1561 aggtaaattg cctgttccgt ggccgacttt ggtgacgact ctgacctatg gtgtccagtg
     1621 tttctctcgg tatccagacc atatgaagca gcatgacttc ttcaagagcg cgatgccaga
     1681 gggttacatt caagaaagga ctatattctt taaagatgac ggcaattaca agactcgcgc
     1741 tgaagttaaa ttcgaaggtg atacattggt aaaccgaata gaacttaagg gtatagactt
     1801 caaagaagat ggaaatatac tgggccacaa acttgagtac aacaaccacg atcagaagga
     1861 cagaaatttg cgacggatca cgcggctcgt gctcgttgtg gttgctgttt ttgttggttg
     1921 ttggaccccg gttcaggtct ttgtgttggc ccaaggtctc ggggtacaac ccagctcaga
     1981 aacagctgtc gcgatattga ggttttgtac ggcgcttgga tacgtcaact catgcttgaa
     2041 ccctatactg tacgctttcc tcgacgaaaa ctttaaaagg tgttttcgag acttctgctt
     2101 cccgttgaag atgcgaatgg agagacaggc cactgcgaga gtgcggaata cggtacagga
     2161 cccggcggcc ctccgagaca tcgacggaat gaacaagcct gtataatgta cagcggccgc
     2221 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2281 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2461 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2521 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2581 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2641 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2701 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2761 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2821 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2881 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3001 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3061 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3121 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3241 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3301 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3361 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3421 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3481 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3541 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3601 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3661 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3721 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3781 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3841 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3901 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3961 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4021 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4081 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4141 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4201 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4261 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4321 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4381 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4441 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4501 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4561 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4621 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4681 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4741 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4801 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4861 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4921 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4981 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5041 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5101 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5161 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5221 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5281 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5341 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5401 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5461 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5521 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
     5581 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     5641 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     5701 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     5761 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     5821 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     5881 atctacgtat tagtcatcgc tattaccatg
//
