LOCUS       sequence-395157-        6232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express muLight biosensor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6232)
  AUTHORS   Dong C, Gowrishankar R, Jin Y, He XJ, Gupta A, Wang H, Sayar-Atasoy 
            N, Flores RJ, Mahe K, Tjahjono N, Liang R, Marley A, Or Mizuno G, Lo
            DK, Sun Q, Whistler JL, Li B, Gomes I, Von Zastrow M, Tejeda HA, 
            Atasoy D, Devi LA, Bruchas MR, Banghart MR, Tian L
  TITLE     Unlocking opioid neuropeptide dynamics with genetically encoded 
            biosensors.
  JOURNAL   Nat Neurosci. 2024 Jul 15. doi: 10.1038/s41593-024-01697-1.
  PUBMED    39009835
REFERENCE   2  (bases 1 to 6232)
  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 
            Neurosci. 2024 Jul 15. doi: 10.1038/s41593-024-01697-1."
COMMENT     Sequence Label: pCMV-µLight
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        178..481
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        482..685
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     636..656
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1861..1882
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1988..2009)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2249..2268)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3137..3258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3174..3193)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3228..3247
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3265..3720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3402..3423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3614..3633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3747..3851
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3853..4210
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3853..4044
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3876..3896)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4061..4196
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4245..5039
                     /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             4245..5039
                     /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(4299..4318)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4909..4928
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5227..5246)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5271..5318
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5647..6232,1..3)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6136..6155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
       61 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      121 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      181 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      241 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      301 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      361 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      421 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      481 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      541 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      601 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      661 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
      721 gactcagatc tcgagcttaa gcttaccggt gcgcgccatg aagacgatca tcgccctgag
      781 ctacatcttc tgcctggtgt tcgccgacta caaggacgat gatgacgcca tggatagtag
      841 cgctgcgcct accaacgcgt caaactgcac cgatgctctt gcgtactcct cctgctcccc
      901 ggcacctagt cccggttctt gggtcaattt gtcccatctg gacggaaacc tctctgatcc
      961 ctgtgggcct aacaggacgg acctcggtgg gagggactcc ctttgcccgc cgaccggatc
     1021 tccgtccatg ataacggcca ttacaattat ggcgttgtat agcatcgtat gcgttgtagg
     1081 tctttttggg aatttcctgg tgatgtacgt catcgtcagg tacacaaaga tgaaaacagc
     1141 tactaacatt tatatattta acctggcgct cgcggacgct ctcgcaacgt caacgctccc
     1201 gtttcagtcc gtgaattatc tcatgggtac ttggcctttc ggaacaatac tgtgtaaaat
     1261 tgttataagc atagattatt ataatatgtt cacgtccatc ttcacactct gcacaatgtc
     1321 tgtggatagg tacattgctg tatgtcaccc acagaaggcg cttgacttta gaactccacg
     1381 caatgcaaag attataaatg tgtgcaactg gatcttgtcc tctgcaatag ggcttcctgt
     1441 gatgttcatg gcgactacta agtacagaca gggcagcata gattgcacac tcaccttctc
     1501 acacccaact tggtactggg aaaatctgct caagatctgc gtcttcattt ttgcttttat
     1561 catgccagta ttgataatca cggtctgtta cgggttgatg attttgcggc tcaaatcagt
     1621 tagaatgttg tctgggagcc tgagctcatg cattaacgtc tatatcaagg ccgacaagca
     1681 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     1741 gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1801 caaccactac ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga agcgcgatca
     1861 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1921 caagggcggt accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt
     1981 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2041 gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2101 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2161 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2221 catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2281 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2341 ggacggcaac atcctggggc acaagctgga gtacaacagc catgaccaac tgaaggagaa
     2401 ggaccgcaac ctcagaagga taacgagaat ggtgctggtc gtagtggcgg ttttcattgt
     2461 ttgttggacg ccaatacaca tatacgtgat tataaaggct ctggtgacaa ttcccgaaac
     2521 aacgtttcag acggtctctt ggcatttctg tattgcattg gggtacacta attcctgcct
     2581 taatcctgta ttgtacgcct ttctggatga aaactttaaa agatgtttcc gcgagttctg
     2641 cataccgacc agcagcaaca ttgaacaaca aaactccacg cgcatacggc aaaatactag
     2701 ggatcacccg tccactgcga atactgtaga ccgaacgaac catcagttgg agaatttgga
     2761 agcggaaact gctcctctgc cacagtccca gcccatcctc aacaccaagg agatggcccc
     2821 gcagagcaag cctccagagg agctggagat gagcagcatg cccagccccg tggcccctct
     2881 gcccgcacgc acggagggcg tcatcgacat gcggagcatg tccagcattg acagcttcat
     2941 cagctgtgcc acggacttcc ctgaagccac cagattcttc tgctacgaga acgaggtgtg
     3001 acaacctgat gtacagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     3061 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3121 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3241 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3301 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3361 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3421 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3481 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3541 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3601 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3661 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3721 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3781 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3841 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3901 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3961 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4021 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4081 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4141 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4201 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4261 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4321 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4381 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4441 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4501 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4561 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4621 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4681 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4741 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4801 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4861 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4921 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4981 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5041 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5101 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5161 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5221 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5281 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5341 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5401 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5461 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5521 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5581 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat gg
//
