LOCUS       sequence-427266-        5935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses norepinephrine indicator nLightR under mammalian promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5935)
  AUTHORS   Kagiampaki Z, Rohner V, Kiss C, Curreli S, Dieter A, Wilhelm M, 
            Harada M, Duss SN, Dernic J, Bhat MA, Zhou X, Ravotto L, Ziebarth T,
            Wasielewski LM, Sonmez L, Benke D, Weber B, Bohacek J, Reiner A, 
            Wiegert JS, Fellin T, Patriarchi T
  TITLE     Sensitive multicolor indicators for monitoring norepinephrine in 
            vivo.
  JOURNAL   Nat Methods. 2023 Sep;20(9):1426-1436. doi: 
            10.1038/s41592-023-01959-z.
  PUBMED    37474807
REFERENCE   2  (bases 1 to 5935)
  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"; doi: "10"; journalName: 
            "Nat Methods"; date: "2023-09"; volume: "20"; issue: "9"; pages: 
            "1426-1436"
COMMENT     Sequence Label: pCMV_nLightR
FEATURES             Location/Qualifiers
     source          1..5935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     236..256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2440..2561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2477..2496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2531..2550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2568..3023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2705..2726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2917..2936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3050..3154
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3156..3513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3156..3347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3179..3199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3364..3499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3548..4342
                     /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             3548..4342
                     /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(3602..3621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4212..4231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4530..4549)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4574..4621
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4950..5538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5439..5458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5713..5935,1..81)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
       61 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      121 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      181 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      241 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      301 cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttggatcc gccaccatga
      361 agacgatcat cgccctgagc tacatcttct gcctggtgtt cgccgactac aaggacgatg
      421 atgacgcagt ctttctcagt gggaacgcta gtgactcttc aaattgtacc cacccaccac
      481 ctcctgttaa tatcagcaag gctattttgc tgggtgtcat cttgggaggc ctgattctgt
      541 tcggagtgct tggtaacatt ctcgtaattc tctcagtagc gtgtcaccgc catctccaca
      601 gcgttaccca ctactatatt gtgaacctcg ctgtagcaga cctgcttctc acgagtacgg
      661 tactgccatt ttccgctatc ttcgaaattt tggggtattg ggcctttgga cgggtttttt
      721 gtaacatctg ggccgctgtg gacgtattgt gttgtacggc atcaatcatg ggcctgtgcc
      781 ttatctcaat agatcgatac tgggcgatta gttcacccga gcgctacgag agaaagatga
      841 ctccgaaggc ggcgttcatt gctctccttt gtgtctgggc tctttccttg gttatatcca
      901 ttgggccctt gttcggctgg cgccagcccg ccccggagga cgaaacaata tgtcagatca
      961 acgaagagcc tggatacgtc cttttcagcg ccctcggatc tttttatgtc ccgttgacaa
     1021 tcatcctggt catgtattgc agagtttatc ggatcgcgca gaaacagctc tccagcttcc
     1081 acgtgagcga gaggatgtat cctgaggacg gtgcgctgaa atccgaaata aagaaaggcc
     1141 tgcgcctcaa agacgggggc cactacgccg ctgaggttaa gacaacttac aaggctaaaa
     1201 agcctgtcca gctcccgggc gcctacatag tcgacattaa actggacata gtcagccata
     1261 acgaggacta caccatagtc gaacagtgtg agcgggcgga aggtcgccac agtacaggag
     1321 gaatggacga attgtataaa gggggcaccg gtggcagctt ggttagcaag ggagaggaag
     1381 ataacatggc tattataaag gagttcatga gatttaaagt tcacatggag ggatctgtaa
     1441 acgggcatga gtttgagatt gagggagaag gagaagggcg accgtacgag gcgttccaaa
     1501 ccgcgaaatt gaaggtcacc aagggaggac cccttccgtt tgcttgggac attcttagcc
     1561 cacaattcat gtatggcagc aaggcgtaca taaagcaccc ggcagacata cccgactact
     1621 tcaaacttag ctttccagag ggtttccgat gggaaagagt gatgaacttt gaggatggtg
     1681 gcatcataca cgtcaatcaa gatagcagcc ttcaggacgg agtatttata tacaaagtca
     1741 aactccgggg tacaaacttt cccccggacg ggccagtcat gcagaagaag actatgggtt
     1801 gggaggcgcg gaaagacgac ttgaagaggg agactaaagt cctcaagacg ctcgggatag
     1861 ttgttggttg tttcgtactc tgctggttgc cctttttctt ggtgatgccg ataggtagct
     1921 tttttcccga cttccgaccg agtgagacag tattcaaaat agctttttgg cttggttatt
     1981 tgaactcctg tattaatccc attatatatc cctgttcatc tcaagagttt aaaaaggctt
     2041 ttcaaaacgt gttgcgcata caatgcctgc gcagaaagca gtcttctaag cacgcactcg
     2101 gttatacgct ccacccccca tcccatgctc tcgagggaca gtatagaaac ctcgtgcgaa
     2161 tacctgtggg tagtggcgaa acattttgca aaatcagtaa aacagacggg gtatgtgaat
     2221 ggaaattttt ttcatccttg ccacgcggat ctgccagaat aacagttcct aaggaccagt
     2281 ccgcttgcac aactgcccgg gacttttaag aattcagagc ggccgcgact ctagatcata
     2341 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2401 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2521 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2581 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2641 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2701 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2761 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2821 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2881 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2941 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3001 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3061 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3121 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3241 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3301 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3361 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3421 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcaa gaagtagtga
     3481 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3541 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3601 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3661 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3721 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3781 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3841 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3901 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3961 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4021 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4081 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4141 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4201 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4261 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4321 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4381 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4441 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4501 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4561 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4621 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4681 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4741 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4801 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4861 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4921 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4981 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5041 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5101 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5161 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5221 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5281 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5341 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5401 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5461 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5521 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5581 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5641 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5701 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgccaaacga
     5761 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5821 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5881 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcc
//
