LOCUS       sequence-215118-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nLight3.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  AUTHORS   Patriarchi T, Cho JR, Merten K, Howe MW, Marley A, Xiong WH, Folk 
            RW, Broussard GJ, Liang R, Jang MJ, Zhong H, Dombeck D, von Zastrow 
            M, Nimmerjahn A, Gradinaru V, Williams JT, Tian L
  TITLE     Ultrafast neuronal imaging of dopamine dynamics with designed 
            genetically encoded sensors.
  JOURNAL   Science. 2018 Jun 29;360(6396). pii: science.aat4422. doi: 
            10.1126/science.aat4422. Epub 2018 May 31.
  PUBMED    29853555
REFERENCE   2  (bases 1 to 5739)
  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: "Science."; 
            date: "2018-06-29"; volume: "360(6396). pii: science.aat4422. doi"; 
            pages: " 10.1126/science.aat4422. Epub 2018 May 31"
COMMENT     Sequence Label: pCMV-nLight3.1
FEATURES             Location/Qualifiers
     source          1..5739
                     /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"
     primer_bind     1252..1273
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1379..1400)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1640..1659)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2163..2284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2200..2219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2254..2273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2291..2746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2428..2449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2640..2659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2773..2877
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2879..3236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2879..3070
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2902..2922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3087..3222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3271..4065
                     /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             3271..4065
                     /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(3325..3344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3935..3954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4253..4272)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4297..4344
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4673..5261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5162..5181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5436..5739
                     /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 acgccatggg
      361 ctccctccag ccggacgcag gcaatgcgtc atggaatggg acagaagccc ctggcggagg
      421 cgcaagggca acgccatata gtcttcaagt aaccctgact cttgtctgcc tcgcaggtct
      481 cctgatgctc ctgacggtct ttggcaatgt gcttgtgata atagctgtat ttacatccag
      541 ggcactcaag gcgccacaaa acttgttttt ggtttcactg gcgtccgccg acattcttgt
      601 ggctacgctc gtcatcccct tctcactcgc taatgaagtg atgggttatt ggtactttgg
      661 caaggcgtgg tgtgagatat atcttgcact cgatgtcttg ttctgcacga gcagtatagt
      721 tcatctctgc gctatctcat tggatagata ttggtcaata acgcaagcta ttgagtataa
      781 cctgaagaga acccctagac ggatcaaggc cattattatt accgtttggg tcatatcagc
      841 ggtcatctca ttccctccct tgataagcat tgagaaaaaa ggtggaggcg gaggaccgca
      901 acctgctgag cccagatgtg agataaacga ccaaaaatgg tacgttatat caagttgtat
      961 cgggtcattc tttgcgccat gcttgattat gattctggta tatgttagaa tctatcagat
     1021 agcgaaacgg ctgagctcac tcattaacgt ctatatcaag gccgacaagc agaagaacgg
     1081 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccta
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1201 cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt ataagggcgg
     1321 tactggaggg agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1381 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg
     1441 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1501 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1561 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acatccagga
     1621 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1681 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1741 catcctgggg cacaagctgg agtacaacaa tcatgaccaa ctgagggaga aacgatttac
     1801 gttcgttctc gccgtagtca tcggtgtttt cgtagtttgc tggttccctt tcttttttac
     1861 atacacattg actgccgtcg gctgttctgt accccgaaca ctttttaaat tttttttctg
     1921 gttcgggtac tgcaactcta gccttaatcc ggtgatatac acaattttta atcatgattt
     1981 tcggcgggct tttaagaaaa ttctctgtcg gggggaccga aaacgcatag tgtagtgtac
     2041 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2101 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2161 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2221 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2281 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2341 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2401 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2461 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2521 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2581 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2641 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2701 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2761 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2821 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2881 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3121 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3181 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3241 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3421 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3781 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4201 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4261 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4321 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4381 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4441 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4501 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4561 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5341 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     5401 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5461 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5521 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5581 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5641 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5701 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
