LOCUS       sequence-317300-        5521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondria-targeted genetically encoded fluorescent indicator for 
            measuring NAD/NADH ratio..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5521)
  AUTHORS   Bilan DS, Matlashov ME, Gorokhovatsky AY, Schultz C, Enikolopov G, 
            Belousov VV
  TITLE     Genetically encoded fluorescent indicator for imaging NAD/NADH ratio
            changes in different cellular compartments.
  JOURNAL   Biochim Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519-9. doi: 
            10.1016/j.bbagen.2013.11.018.
  PUBMED    24286672
REFERENCE   2  (bases 1 to 5521)
  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: "Biochim 
            Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519-9. doi: 
            10.1016/j.bbagen.2013.11.018."
COMMENT     Sequence Label: pC1-mitoRexYFP
FEATURES             Location/Qualifiers
     source          1..5521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             538..612
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             634..705
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     1203..1224
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1333..1354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1594..1613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2250..2371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2287..2306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2341..2360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2378..2833)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2515..2536)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2727..2746
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2860..2964
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2966..3323
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2966..3157
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2989..3009)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3174..3309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3358..4152
                     /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             3358..4152
                     /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(3412..3431)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4022..4041
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4340..4359)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4384..4431
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4760..5348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5249..5268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcatg
      541 tccgtcctga cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg
      601 ccgcgcgcca agatccattc gttgggggat ctgtccgtcc tgacgccgct gctgctgcgg
      661 ggcttgacag gctcggcccg gcggctccca gtgccgcgcg ccaagatcca ttcgttgggg
      721 gatccaccgg tcgccaccat gaaggtcccc gaagcggcca tcagccgcct gatcacctac
      781 ctccgcatcc tggaggagct ggaggcccag ggggttcacc ggacgagctc cgagcagctt
      841 ggggagctcg cccaggtcac ggccttccag gtgcggaagg acctttccta cttcggctcc
      901 tacgggacgc gcggggtggg gtacaccgtc cccgtcctga agcgggagct ccgccacatc
      961 ctggggctca accgctccgc cggctacaac agcgacaacg tctatatcat ggccgacaag
     1021 cagaagaacg gcatcaaggc caacttcaag atccgccaca acgtcgagga cggcagcgtg
     1081 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1141 gacaaccact acctgagctt ccagtccgtc ctgagcaaag accccaacga gaagcgcgat
     1201 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcccggcat ggacgagctg
     1261 aacaacgtgg atggcggtag cggtggcacc ggcagcaagg gcgaggagct gttcaccggg
     1321 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1381 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1441 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tcggctacgg cctgaagtgc
     1501 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1561 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1621 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcggcttc
     1681 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acggcaccaa gtggggcctg
     1741 tgcatcgtgg gcatgggccg gctcgggagc gccctggccg actacccggg gttcggggag
     1801 agctttgagc tcaggggctt ctttgacgtg gacccggaga aggtgggccg cccggtgcgg
     1861 ggcggggtca ttgagcacgt ggacctcctg ccccagcggg ttcccggccg cattgagatc
     1921 gccctcctca ccgtgccccg ggaagcggcc cagaaggcgg cggacctgtt ggtggcggcg
     1981 gggatcaagg ggatcctgaa cttcgccccg gtggtcctcg aggtgcccaa ggaggtggcg
     2041 gtggagaacg tggacttctt ggcggggctc acgcggcttt ctttcgctat cctgaacccc
     2101 aagtggcggg aagagatgat ggggtgaagc ggccgcgact ctagatcata atcagccata
     2161 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2221 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2281 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2341 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2401 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2461 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2521 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2581 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2641 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2701 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2761 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2821 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2881 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2941 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3001 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3061 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3121 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3181 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3241 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3301 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3361 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3421 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3481 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     3541 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3601 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3661 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3721 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3781 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3841 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     3901 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3961 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4021 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4081 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4141 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4201 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4261 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4321 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4381 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4441 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4501 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     4561 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     4621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5401 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     5461 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     5521 c
//
