LOCUS       sequence-375341-        5547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ATP measurement, mitochondria outer membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5547)
  AUTHORS   Yaginuma H,  Okada Y
  TITLE     Live cell imaging of metabolic heterogeneity by quantitative 
            fluorescent ATP indicator protein, QUEEN-37C
  JOURNAL   bioRxiv 2021.10.08.463131
REFERENCE   2  (bases 1 to 5547)
  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: "bioRxiv 
            2021.10.08.463131"
COMMENT     Sequence Label: pN1-TOMM20-QUE37C
FEATURES             Location/Qualifiers
     source          1..5547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1368..1619
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1556..1577
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1683..1704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1944..1963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2275..2396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2312..2331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2366..2385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2403..2858)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2540..2561)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2752..2771
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2885..2989
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2991..3348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2991..3182
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3014..3034)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3199..3334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3383..4177
                     /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             3383..4177
                     /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(3437..3456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4047..4066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4365..4384)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4409..4456
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4785..5373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5274..5293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      541 ccatggtggg tcggaacagc gccatcgccg ccggtgtatg cggggccctt ttcattgggt
      601 actgcatcta cttcgaccgc aaaagacgaa gtgaccccaa cttcaagaac aggcttcgag
      661 aacgaagaaa gaaacagaag cttgccaagg agagagctgg gctttccaag ttacctgacc
      721 ttaaagatgc tgaagctgtt cagaagttct tccttgaaga aatacagctt ggtgaagagt
      781 tactagctca aggtgaatat gagaagggcg tagaccatct gacaaatgca attgctgtgt
      841 gtggacagcc acagcagtta ctgcaggtct tacagcaaac tcttccacca ccagtgttcc
      901 agatgcttct gactaagctc ccaacaatta gtcagagaat tgtaagtgct cagagcttgg
      961 ctgaagatga tgtggaaggc ggtagcgggg atccaccggt cgccaccatg gtgaaaactg
     1021 tgaaagtgaa tataacaacc cctgatgggc cagtctacga cgctgatatc gagatggtgt
     1081 ccgtgcgggc cgagagtggt gatctcggca tcctccccgg tcacattccc acaaaggccc
     1141 cactgaagat cggagctgtg cggctgaaga aggacggcca aacagaggat gtcgcagtct
     1201 caggcggcac tgttgaagtg cggcctgacc acgttaccat taatgctcaa gccgctgaac
     1261 gggcggagga cattgacgtc ctccgcgcca aagcggcgaa agagcgggcg gagcgccgcc
     1321 tgcaaagcca gacccgttac aacagccaca acgtctatat catggccgac aagcagaaga
     1381 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1441 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1501 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1561 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     1621 gtggcaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1681 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1741 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1801 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1861 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1921 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1981 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2041 gcaacatcct ggggcacaag ctggagtaca acctgggtga catcgacttc aaacgggccg
     2101 aactggcgtt aaaacgcgcc atgaaccgtt tgagcgttgc ggaagataag tgagcggccg
     2161 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2221 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2281 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2341 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2401 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2461 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2521 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2581 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2641 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2701 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2761 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2821 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2881 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2941 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3001 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3361 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3421 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3481 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3541 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3601 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3661 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3721 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3781 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3841 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3901 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3961 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4021 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4081 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4141 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4201 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4261 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4321 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4381 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4441 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4501 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4561 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4621 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4681 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4741 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4801 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4861 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4921 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4981 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5041 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5101 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5161 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5221 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5281 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5341 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5401 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5461 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     5521 agttcatagc ccatatatgg agttccg
//
